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Objective: JWH133, a cannabinoid type 2 receptor agonist, was tested for its ability to protect mice from bleomycin-induced pulmonary fibrosis. Methods: By using a random number generator, 24 C57BL/6J male mice were randomly divided into the control group, model group, JWH133 intervention group, and JWH133+a cannabinoid type-2 receptor antagonist (AM630) inhibitor group, with 6 mice in each group. A mouse pulmonary fibrosis model was established by tracheal instillation of bleomycin (5 mg/kg). Starting from the first day after modeling, the control group mice were intraperitoneally injected with 0.1 ml of 0.9% sodium chloride solution, and the model group mice were intraperitoneally injected with 0.1 ml of 0.9% sodium chloride solution. The JWH133 intervention group mice were intraperitoneally injected with 0.1 ml of JWH133 (2.5 mg/kg, dissolved in physiological saline), and the JWH133+AM630 antagonistic group mice were intraperitoneally injected with 0.1 ml of JWH133 (2.5 mg/kg) and AM630 (2.5 mg/kg). After 28 days, all mice were killed; the lung tissue was obtained, pathological changes were observed, and alveolar inflammation scores and Ashcroft scores were calculated. The content of type Ⅰ collagen in the lung tissue of the four groups of mice was measured using immunohistochemistry. The levels of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) in the serum of the four groups of mice were measured using enzyme-linked immunosorbent assay (ELISA), and the content of hydroxyproline (HYP) in the lung tissue of the four groups of mice was measured. Western blotting was used to measure the protein expression levels of type Ⅲ collagen, α-smooth muscle actin (α-SMA), extracellular signal regulated kinase (ERK1/2), phosphorylated P-ERK1/2 (P-ERK1/2), and phosphorylated ribosome S6 kinase type 1 (P-p90RSK) in the lung tissue of mice in the four groups. Real-time quantitative polymerase chain reaction was used to measure the expression levels of collagen Ⅰ, collagen Ⅲ, and α-SMA mRNA in the lung tissue of the four groups of mice. Results: Compared with the control group, the pathological changes in the lung tissue of the model group mice worsened, with an increase in alveolar inflammation score (3.833±0.408 vs. 0.833±0.408, P <0.05), an increase in Ashcroft score (7.333±0.516 vs. 2.000±0.633, P <0.05), an increase in type Ⅰ collagen absorbance value (0.065±0.008 vs. 0.018±0.006, P <0.05), an increase in inflammatory cell infiltration, and an increase in hydroxyproline levels [(1.551±0.051) μg/mg vs. (0.974±0.060) μg/mg, P <0.05]. Compared with the model group, the JWH133 intervention group showed reduced pathological changes in lung tissue, decreased alveolar inflammation score (1.833±0.408, P <0.05), decreased Ashcroft score (4.167±0.753, P <0.05), decreased type Ⅰ collagen absorbance value (0.032±0.004, P <0.05), reduced inflammatory cell infiltration, and decreased hydroxyproline levels [(1.148±0.055) μg/mg, P <0.05]. Compared with the JWH133 intervention group, the JWH133+AM630 antagonistic group showed more severe pathological changes in the lung tissue of mice, increased alveolar inflammation score and Ashcroft score, increased type Ⅰ collagen absorbance value, increased inflammatory cell infiltration, and increased hydroxyproline levels. Compared with the control group, the expression of α-SMA, type Ⅲ collagen, P-ERK1/2, and P-p90RSK proteins in the lung tissue of the model group mice increased, while the expression of type Ⅰ collagen, type Ⅲ collagen, and α-SMA mRNA increased. Compared with the model group, the protein expression of α-SMA (relative expression 0.60±0.17 vs. 1.34±0.19, P <0.05), type Ⅲ collagen (relative expression 0.52±0.09 vs. 1.35±0.14, P <0.05), P-ERK1/2 (relative expression 0.32±0.11 vs. 1.14±0.14, P <0.05), and P-p90RSK (relative expression 0.43±0.14 vs. 1.15±0.07, P <0.05) decreased in the JWH133 intervention group. The type Ⅰ collagen mRNA (2.190±0.362 vs. 5.078±0.792, P <0.05), type Ⅲ collagen mRNA (1.750±0.290 vs. 4.935±0.456, P <0.05), and α-SMA mRNA (1.588±0.060 vs. 5.192±0.506, P <0.05) decreased. Compared with the JWH133 intervention group, the JWH133+AM630 antagonistic group increased the expression of α-SMA, type Ⅲ collagen, P-ERK1/2, and P-p90RSK protein in the lung tissue of mice, and increased the expression of type Ⅲ collagen and α-SMA mRNA. Conclusion: In mice with bleomycin-induced pulmonary fibrosis, the cannabinoid type-2 receptor agonist JWH133 inhibited inflammation and improved extracellular matrix deposition, which alleviated lung fibrosis. The underlying mechanism of action may be related to the activation of the ERK1/2-RSK1 signaling pathway.

Objective: To investigate the effect of hydronidone on CCl 4 -induced liver fibrosis in rats and explore the possible mechanism., Methods: Sixty-six male SD rats were randomized into 5 groups, including a control group ( n =10), a liver fibrosis model group ( n =20), 2 hydronidone dose groups (100 and 250 mg/kg; n =12), and a pirfenidone (250 mg/kg) treatment group ( n = 12). Rat models of liver fibrosis were established by subcutaneous injection of CCl 4 in all but the control group. Hydronidone and pirfenidone were given daily at the indicated doses by intragastric administration for 6 weeks. After the treatments, serum samples were collected from the rats for detecting liver function parameters, and hydroxyproline content in the liver tissue was determined. Inflammation and fibrosis in the liver tissue were observed using HE staining and Sirius Red staining. In the cell experiment, human hepatic stellate cell line LX-2 was stimulated with TGF-β1 and treated with hydronidone or pirfenidone, and the expression levels of α-SMA, collagen type I and phosphorylated Smad3, phosphorylated p38, phosphorylated ERK1/2 and phosphorylated Akt were detected with Western blotting., Results: In the rat models of liver fibrosis, treatment with hydronidone obviously improved the liver functions, reduced the content of hydroxyproline in the liver tissue, and significantly alleviated liver fibrosis ( P < 0.05). In LX-2 cells, hydronidone dose-dependently decreased the expression levels of α-SMA and collagen type I. In TGF- β1-stimulated cells, the phosphorylation levels of Smad3, P38, ERK, and Akt increased progressively with the extension of the treatment time, but this effect was significantly attenuated by treatment with hydronidone ( P < 0.05)., Conclusion: Hydronidone can inhibit the phosphorylation of the proteins in the TGF-β signaling pathway, thereby preventing TGF-β1-mediated activation of hepatic stellate cells, which may be a possible mechanism by which hydronidone alleviates CCl 4 -induced liver fibrosis in rats.

Objective: To investigate the effects of Coriaria Sinica Maxim's extract(CSME) on microcirculation and oxidative stress of wounds in rats with deep second-degree burn., Methods: One hundred and eighty rats were randomly divided into normal saline group(NS), white petroleum group(WPL), silver sulfadiazine group (SSD), Coriariasinica Maxim's extract group which were divided into low dose(CSME-L),middle dose(CSME-M) and high dose(CSME-H). After anesthesia with burn instrument to burn the hair removal area of rats, these wounds were confirmed by pathological results with deep second degree burns.And then,those drugs were applied respectively on the wounds,such as NS、WPL、SSD and different concentrations of CSME. After injury at 48 h, 7 d, 14 d and 21 d,the healing rate(HR) of wound was measured, and the microvessel density (MVD), tissue moisture (TM), vascular endothelial growth factor (VEGF), model driven architecture (MDA), superoxide dismutase(SOD) and hydroxyproline(HYP) were detected, too. All pathological sections of the wound tissue were observed., Results: The HR of CSME groups were obviously increased with a dose-dependent manner, which was significantly higher than that of NS and WPL ( P <0.05); On the 21 st day, the diameter, number, distribution of the vessels and and the TM were less than other groups with a dose-dependent manner; On the 7 th and 14 th day after injury, CSME groups were significantly higher than the NS, WPL and SSD with a dose-dependent manner ( P <0.05), but, on the 21 st day after injury, they were lower than NS, WPL and SSD with a dose-dependent ( P <0.05) manner. The levels of SOD, HYP, NO and ET in CSME groups were higher than those in other groups with dose-dependent on SOD activity, HYP, NO and ET content ( P <0.05), while MDA activity was weaker than other groups ( P <0.05). Similarly, pathological findings were also shown that CSME groups were better than other groups with a dose-dependent manner in decrease decreasing of wound repair time and hyperplasia of scar tissue., Conclusions: CSME can relieve tissue edema, promote wound contraction, speed up the formation of eschar and accelerate the proliferation of granulation tissue, which are beneficial to the wound healing in the early stages. But, it can inhibit the hyperplasia of granulation tissue to prevent the excessive scar hyperplasia of burn wound in the later stages. Its mechanism is related to regulation what microcirculation, oxidativestress, NO and VEGF.

Objective: To investigate the effect of edaravone on oxidative stress and myocardial fibrosis induced by isoproterenol in rats., Methods: Fifty male SD rats were randomly divided into 5 groups, including a control group, a myocardial fibrosis model (established by injections of isopropyl adrenaline for 10 days) group, and 3 edaravone groups with edaravone treatment at low, medium, or high doses for 14 days. After the treatments, the rats were examined for the degree of myocardial fibrosis, left ventricular mass index (LVMI), collagen volume fraction (CVF), and myocardial contents of collagen I (Col I), collage III (Col III), hydroxyproline (Hyp), superoxide dismutase (SOD), malondialdehyde (MDA), and nitric oxide (NO); The expression of transforming growth factor-β1 (TGF-β1) in the myocardial tissues was examined by immunofluorescence assay and Western blotting., Results: Compared with the control rats, the rat models of myocardial fibrosis showed significantly increased CVF and LVMI (P=0.000), which were lowered by edaravone treatments in a dose-dependent manner (P<0.05). The myocardial contents of Col I, Col III and Hyp also increased in the model group (P=0.000) and were lowered dose-dependently by edaravone; the contents of MDA was higher (P=0.000) and SOD and NO were lower in the model group (P=0.000), and edaravone treatments obviously increased SOD and NO contents (P<0.05). The model rats showed significantly increased myocardial expression of TGF-β1 (P=0.000), which was markedly lowered by edaravone treatments (P=0.000). The myocardial content of MDA was positively correlated while SOD and NO were negatively with LVMI, CVF, Col I, Col III and Hyp; TGF-β1 was positively correlated with LVMI, CVF, Col I, Col III, Hyp and MDA but negatively with SOD and NO., Conclusion: Edaravone can relieve oxidative stress and inhibit TGF-β1 activation to ameliorate myocardial fibrosis in rats.

Objective: To compare the effects of oral treatment with tetrandrine (TD) and N-acetylcys-teine (NAC) separately or jointly on silica-exposed rats., Methods: 40 sprague-Dawly (SD) rats were randomly divided into normal saline group, quartz group, TD treatment group (50 mg/kg), NAC treatment group (500 mg/kg) and combined treatment group (TD: 50 mg/kg + NAC: 500 mg/kg). Rats in normal saline group and other groups received intratracheal instillation of normal saline and quartz dust suspension respectively. Treatment groups were given TD, NAC separately or jointly via esophagus the next day after instillation, once a day and six times a week for 30 consecutive days. At the end of experiment, the pathological changes of lung tissues were evaluated by the methods of Foot, HE and Masson staining, the level of hydroxyproline (HYP), malondjalde-hyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in lung tissues were measured by alkaline hydrolysis method, the barbituric acid method and enzyme-linked immunosorbent assay (ELISA) respectively., Results: Compared with the quartz group, lymph nodes/body coefficients in all treatment groups and lung/body coefficient in combined treatment group were significantly decreased (P < 0.05). Pathology results showed that the normal saline group demonstrated no obvious evidence of lung damage. The quartz group lungs silicotic lesions focused on II~III level, the TD treatment group was mainly with I level, the NAC treatment group was mainly with I~II level, and the combined treatment group only showed little silicotic nodule, no obvious fibrosis. HYP content in TD treatment group and combined treatment group were significantly lower than that in the quartz group (P < 0.05), while it showed no obvious change in NAC treatment group. MDA content in lung tissues of each treatment group (TD treatment group, NAC treatment group and combined treatment group) were 18.80 ± 2.94, 20.13 ± 4.01 and 17.05 ± 3.52 nmol/ml respectively, which lower than in the quartz group (23.99 ± 3.26 nmol/ml). The level of IL-6 in lung tissues of the quartz group were 89.57 ± 8.78 pg/ml. After TD and NAC monotherapy, the IL-6 content decreased to 79.22 ± 9.65 pg/ml and 81.63 ± 5.72 pg/ml, and it decreased more significantly after combined medication (74.37 ± 3.17 pg/ml). The level of TNF-α in the quartz group were 59.05 ± 4.48 pg/ml. After TD and NAC monotherapy, the TNF-α content decreased to 50.48 ± 2.76 pg/ml and 54.28 ± 4.30 pg/ml, and it decreased more significantly after combined medication (49.10 ± 4.98 pg/ml)., Conclusion: NAC and TD could reduce MDA, TNF-α and IL-6 levels in lung tissue, and alleviate SiO2-induced pulmonary fibrosis in rats. Combined treatment with TD and NAC was more effective than TD or NAC treatment separately.

Objective: To investigate the effects of Artesunate(Art) on the LX-2 cell., Methods: The cultured hepatic stellate cells were divided into control group and Art-treated groups with 250,350,450 µmol/L. The rate of cellular proliferation was detected by MIT assay, the content of ceramide (Cer)was determined by HPLC method, the content of hydroxyproline (Hyp) was determined by enzyme digestion method, the expressions of PPAR-γ, p53 and Caspase 3 were detected by Western blot., Results: Compared with control group, IX-2 treated with Art were inhibited in a concentration-dependent manner(P < 0.01). Art could significantly increase the content of cerarnide in LX-2 ( P <0.01), and the content of Hyp was significantly decreased (P <0.05, P <0.01). The expressions of PPAR-γ, p53 and Caspase 3 were increased compared with that of control group(P < 0.01)., Conclusion: Artesunate could inhibit the proliferation and induce apoptosis of hepatic stellate cells through upregulating ceramide.

Objective: To study the protective effect of alcohol extract of Plumula Nelumbini (AEPN) on carbon tetrachloride (CCl4) induced hepatic fibrosis rats and to explore its possible mechanism., Methods: Totally 32 male SD rats were randomly divided into four groups, i.e., the normal control group, the model group, the high dose AEPN group, and the low dose AEPN group, 8 in each group. 1,000 mg/kg AEPN was given to rats in the high dose AEPN group by gastrogavage at 10 mL/kg, once daily, while 500 mg/kg AEPN was given to rats in the low dose AEPN group by gastrogavage at 10 mL/kg, once daily. Hepatic fibrosis was induced by intraperitoneal injection of CCl4. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and albumin (ALB) were examined using automatic biochemical analyzer. Activities of superoxide dismutase (SOD), contents of malondialdehyde (MDA) and hydroxyproline (Hyp) in the hepatic tissue were determined using colorimetry. The degree of liver fibrosis was observed by HE staining and Masson staining. The expression of α-smooth muscle actin (α-SMA) was detected using immunohistochemistry., Results: (1) Compared with the normal control group, serum levels of ALT and AST obviously increased and the serum ALB level obviously decreased in the model group (all P < 0.05). After treated by AEPN, serum levels of ALT and AST were lowered. and the serum ALB level was higher (all P < 0.05). (2) Compared with the normal control group, collagen deposition was obviously seen in rats' livers of the model group, and pseudolobule had formed; inflammatory activities and fibrosis degrees were serious; contents of Hyp also increased (P < 0.05).After treated by AEPN, collagen deposition was obviously reduced with no obvious pseudolobule; inflammatory activities and fibrosis degrees were alleviated; contents of Hyp were also lowered (P < 0.05). (3) Compared with the normal control group, contents of MDA in the liver tissue obviously increased, while activities of SOD obviously decreased (P < 0.05) in the model group. After treated by AEPN, contents of MDA in the liver tissue decreased and the serum SOD level significantly increased (all P < 0.05). (4) Compared with the normal control group, the expression of α-SMA was obviously elevated in the model group (P < 0.05). After treated by AEPN, its expression was obviously lowered (P < 0.05)., Conclusions: AEPN could fight against CCl4 induced liver fibrosis in rats. Fighting against lipid peroxidation and inhibi- ting activation and proliferation of hepatic stellate cells might be possibly main mechanism.

Objective: To study the pathological process and characteristics of bleomycin (BLM)-induced interstitial lung disease (ILD) model and collagen-induced arthritis (CIA) complicated by BLM-induced ILD (CIA-ILD) model in mice and explore the correlations with circulating fibrocytes., Methods: Ninety mice were randomly divided into normal saline group (S group), BLM group (B group), CIA-BLM group (CB group), with 30 mice in each group. On the 2, 7, 14, 21 and 28 days after BLM challenge, mice of each group were sacrificed. HE staining was used to detect the degrees of acute inflammation and sirius red staining to detect lung fibrosis. Immunohistochemicstry was adopted to detect alpha-smooth muscle actin expression. The peripheral blood was obtained to count the proportion of circulating fibrocytes (CD45⁺COL1⁺) by flow cytometry. The correlations between circulating fibrocytes and pathological changes of ILD were analyzed statistically., Results: Both of the ILD model groups showed the dynamic process from pulmonary alveolitis to fibrosis gradually. Inflammatory lesion in the lungs of B group was the most obvious on the 7-14th day, and then there was a slow rehabilitation process on the 21th and 28th day. CB group began to present alveoli destruction and collagen deposition on the 14th day, and the inflammation was the most severe on the 28th day. Pulmonary hydroxyproline content was obviously ascended in CB group as compared with B group (P<0.05) after 14 days. An increase in the total number of circulating fibrocytes was observed in both B and CB groups. It tended to increase at first, and subsequently declined. The level of circulating fibrocytes was higher in CB group than in B group on day 14, 21 and 28 (P<0.05). The immunohistochemical results showed that there were many myofibroblasts infiltrating in CB group from the 14th day to 28th day. The number of circulating fibrocytes in peripheral blood was positively correlated with pulmonary inflammation, fibrosis degree and hydroxyproline content (r=0.847, 0.826, 0.735, P<0.01)., Conclusion: The pathological process of CIA-ILD animal model is more close to rheumatoid arthritis-induced ILD development. Circulating fibrocytes may be involved in the inflammation and fibrosis progression of lung.

Objective: To observe the expression of leptin (LP) and its influence on pulmonary fibrosis in experimental rats with silicosis and the correlation between the expressions of leptin and HIF-1α., Methods: A total of 120 male Sprague-Dawley rats were randomly divided into normal control group, silicosis model group, LP intervention groups (consisting of LP5, LP10 and LP20 groups according to the concentration of LP). The rats in the normal control group were intratracheally administered 1 mL normal sodium, and the ones in the other groups were intratracheally given 1 mL SiO₂(40 mg/mL) suspension. The rats in LP intervention groups were intraperitoneally injected with leptin 5, 10, 20 ng/kg.d respectively from the first day. Six rats in each group were sacrificed on the 7th, 14th, 21th and 28th day. The expression of LP on the 7th, 14th, 21th and 28th day and the hydroxyproline content on the 28th day in rat lung tissues were measured by ELISA, and the expressions of HIF-1α and LP proteins in the lung tissues of the silicosis model group were measured by Western blotting., Results: There were significant differences in the expression of LP in the lung tissues at each time point (7th, 14th, 21th and 28th day) among the five groups (F=669.18, 948.67, 1 172.00, 521.55, P<0.05). The LP content in the silicosis model group was significantly higher than that in the normal control group (P<0.05) and the expression of LP in the LP intervention groups were significantly elevated at each time point (P<0.05) as compared with the silicosis model group. Hydroxyproline content on 28th day was (0.89 ± 0.16), (3.14 ± 0.40), (3.78 ± 0.27), (4.35 ± 0.13), (4.87 ± 0.16) mg/g in normal control group, silicosis model group, LP5, LP10 and LP20 intervention groups, respectively. Compared with the normal control group, hydroxyproline content in the silicosis model group significantly increased (P<0.05), and compared with the silicosis model group, hydroxyproline content in LP intervention groups were significantly raised (P<0.05). The significant positive correlations were noted between the expressions of HIF-1α and LP (r=0.876, 0.796, 0.859 and 0.854 at each time, P<0.05)., Conclusion: The expression of LP goes up significantly in the lung tissues of rats with silicosis. Addition of exogenous leptin can increase the collagen deposition in the lung tissues; meanwhile leptin has a positive correlation with HIF-1α.

Objective: To discuss the reverse effect of Yinchenhao decoction(YCHD) in dimethyl nitrosamine (DMN)-induced hepatic fibrosis in rats., Method: The rat hepatic fibrosis model was established through the intraperitoneal injection with 1% dimethyl nitrosamine (DMN) with a dose of 1.0 mL x kg(-1) x d(-1) for consecutively three weeks, once for the first three days of each. The rats were randomly divided into six groups: the silymarin positive control group (50.0 mg x kg(-1) x d(-1), YCHD high (20.0 g x kg(-1) d(-1)), middle (8.0 g x kg(-1) x d(-1)) and low (3.2 g x kg(-1) x d(-1)) dose groups, the model group and the normal control group. The model group and the normal control group were orally administered with normal saline for consecutively five weeks. The pathologic changes in liver tissues were observed by HE staining. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), g-glutamyltransferase (g-GGT), hyaluronic acid (HA), laminin (LN), collagen type IV (CIV) and type III procollagen amino terminal peptide (PIIINP) in serum were determined. The metabolite profiling of amino acid and the content of hydroxyproline in liver tissues were also measured., Result: Compared with the model group, YCHD high and middle dose groups could significantly reverse the pathologic changes in liver tissues of rats. YCHD could reduce the levels of ALT, AST, gamma-GGT, HA, LN, CIV, PIIINP in serum and the content of hydroxyproline in liver tissues in a dose-dependent manner, and altered the metabolite profiling of amino acid in rat liver tissues., Conclusion: YCHD has the effect in reversing dimethyl nitrosamine induced hepatic fibrosis in rats.

Objective: To study the effect of Wu-He Dipsacus asper (WHDA), Traditional Chinese Medicine, injection on mice-aging model induced by D-galactose., Methods: Forty-eight Kunming mice (24 male and 24 female) were randomly divided into control group, model group, positive control group, 7.2 g/kg WHDA group, 3.6 g/kg WHDA group and 1.8 g/kg WHDA group with eight in each group. The model was induced through injecting D-galactose into peritoneal cavity and Morris water maze was used to detect the learning and cognitive ability of mice. The skin hydroxyproline, brain tissue malondialdehyde (MDA), lipofuscin (LP), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels of mice were detected; the IL-2 and IL-6 levels in serum of mice were detected by using double antibody sandwich ELISA method., Results: Each WHDA group was significantly reduced in latency period compared with the model group during Morris water maze test (P < 0.05) and the number of mice in model group through the platform was less than other mice in each group (P < 0.05). The levels of MAD and LP of the control group and each WHDA group were less than model group in the detection of heart, brain tissue oxidation index (SOD, MAD, LP and GSH-Px, P < 0.05). The activity of SOD and GSH-Px in the control group and each WHDA group was significantly higher than that in the model group (P < 0.05). The skin hydroxyproline content of mice which had been injected with D-galactose was significantly lower than that in the control group (P < 0.05) and the skin hydroxyproline content of mice of WHDA group was significantly higher than that in the model group (P < 0.05). The IL-2, IL-6 levels in serum of mice in WHDA group were significantly higher than those in the control group and the model group (P < 0.05) and the IL-2, IL-6 levels in serum of mice in the model group were lower than those in the control group (P < 0.05)., Conclusion: The effective constituents of WHDA have a variety of biological activity which can have a good effect on anti-aging by different ways, improving learning and memory function, eliminating free radicals antioxidant, and enhancing the body immunity and other aspects.

Objective: To investigate the effects of Corbrin Shugan capsule on dimethylnitrosamine (DMN)-induced hepatic fibrosis in rats., Methods: Hepatic fibrosis was induced by DMN in AD rats. The serum concentrations of III pro-collagen (III PC),laminin (LN) and tissue inhibitor of metalloproteinase-1(TIMP-1) were determined with ELISA. The concentration of albumin (ALB) in sera and the content of hydroxyproline (Hyp) in liver tissues were determined with chemical colorimetric and HPLC, respectively. The fibrosis area was measured with Motic Med 6.0 digital medical image analysis system., Results: Compared to model group the high-dose (450 mg kg(-1)),mid-dose (270 mg kg(-1)) and low-dose (90 mg kg(-1)) groups of Corbrin Shugan capsule had significantly lower serum content of III PC [34.46 ± 13.95),(36.15 ± 9.46), and (40.58 ± 7.72)ng ml(-1) compared with (49.38 ± 10.95)ng ml(-1),P<0.05 or P<0.01],TIMP-1 [(16.65 ± 4.24),(16.66 ± 4.34),and (18.99 ± 6.05)ng ml(-1) compared with (30.84 ± 14.48)ng ml(-1), P<0.05 or P<0.01], LN [(12.94 ± 4.29), (12.96 ± 3.21),and (15.32 ± 8.00)ng ml(-1) compared with (30.22 ± 17.00)ng ml(-1),P<0.05 or P<0.01] and smaller hepatic fibrosis area [(0.02240 ± 0.01337), (0.02176 ± 0.01460) and (0.02384 ± 0.01405)μm(2) compared with vs (0.03929 ± 0.01732)μm2, P<0.05 or P<0.01]; the high-dose and mid-dose groups of Corbrin Shugan capsule had significantly lower content of Hyp in liver tissues [(0.77 ± 0.09) and (0.81 ± 0.09)μg μmg(-1) compared with (1.06 ± 0.33)μg mg(-1),P<0.05 or P<0.01]; and the high-dose group of Corbrin Shugan capsule significantly increased the content of ALB in sera [(34.02 ± 4.17)g L(-1) compared with (30.25 ± 4.21)g L(-1),P<0.05]., Conclusion: Corbrin Shugan capsule is effective in treatment of DMN-induced hepatic fibrosis in rats.

Objective: To investigate the effect of ursolic acid (UA) on the alloxan-induced myocardial fibrosis in mice and discuss the possible mechanism., Methods: Diabetes was produced by a single injection of alloxan (70 mg/kg, i.v.) in mice. The mice were randomly divided into four groups: normal control group, model group, ursolic acid group (UA, 35 mg/kg, p.o.) and benazepril group (5 mg/kg, p. o.), and continuous administrated for 8 weeks. The blood glucose was measured 24 hours after the last administration. Detected the specific biochemical of myocardial tissue: superoxide dismutase (SOD), malondialdehyde (MDA) and hydroxyproline(HYP). Using masson staining to observe the morphology of the myocardial tissue. Immunohistochemistry was employed to determine the protein levels of TGF-beta1., Results: Compared to normal group, the blood glucose, heart index, myocardial tissue MDA, HYP level were increased, and SOD activities were decreased in the diabetic mice, Masson stain showed that myocardial cells disarranged, myocardial collagen fibrosis hyperplasia. Meanwhile, the protein expression of TGF-beta1 was increased in model group. The UA group improved all the above significantly., Conclusion: UA improves the myocardial collagen fibrosis in diabetic mice induced by alloxan, its mechanism may be related to inhibiting the expression of TGF-beta1 and antioxidation.

Objective: To investigate the effect of the anti-laminin receptor 1 (anti-LAMR1) monoclonal antibody (mAb) on rat pulmonary fibrosis induced by bleomycin., Methods: Seventy-two male Wistar rats were randomly divided into 3 groups: LAMR1 mAb group (group L), control group (group C) and model group (group M). All rats were injected with bleomycin via tracheal instillation (5 mg/kg body mass) to induce pulmonary fibrosis. Then, the rats were treated with anti-LAMR1 mAb (group L), dexamethasone (group C), normal saline (group M), three times each week via intraperitoneal injection. Eight rats in each group were sacrificed at the 7, 14, 28 days. Histological changes of the lungs were evaluated by HE staining. The expression of surfactant protein A (SP-A) in the lung tissue was measured by immunohistochemistry. The expression of hydroxyproline (Hyp) in serum was measured by ELISA. The expressions of MMP-9 and TGF-β1 mRNA were evaluated by PCR., Results: Compared with the other two groups, group M presented more apparent alveolitis and the higher expression of hydroxyproline in serum, the lower expression of SP-A, and the higher expressions of MMP-9 and TGF-β1 mRNA in the lung tissue., Conclusion: LAMR1 mAb can evidently alleviate pulmonary fibrosis induced by bleomycin in rats.

Objective: To study the effects of iptakalim (Ipt), an ATP-sensitive potassium channel opener, on cardiac remodeling induced by isoproterenol (ISO) in Wistar rats., Methods: ISO was given subcutaneously (85 mg/(kg x d), sc, 7 days) to induce cardiac remodeling in rats. The rats in Ipt treated group were administrated with Ipt 3 mg/kg (po) after ISO injection. After treated with Ipt for 6 weeks, the hemodynamic parameters were tested by an eight channel physiological recorder (RM-6000). Then the heart weight was weighed and the cardiac remodeling index was calculated. HE stain and Masson's stain were employed to perform histological analysis, the hydroxyproline(Hyp) content in cardiac tissue was detected by colorimetric method, radioimmunoassay was used to measure the plasma levels of endothelin-1 (ET-1) and prostacyclin (PGI2)., Results: Six weeks after ISO injection, the cardiac functions of model group were damaged markedly compared with those of normal group. The characteristics of ventricular remodeling in model group included that the heart weight index, myocyte cross-sectional area, myocardial fibrosis, and the hydroxyproline content in cardiac tissue were all increased significantly. The plasma level of ET-1 was increased, while the plasma level of PGI2 was decreased significantly. These changes could be reversed by Ipt treatment (3 mg/(kg x d) for 6 weeks)., Conclusion: Ipt can reverse cardiac remodeling induced by isoproterenol in rats. The endothelial protective effect regulating effects of Ipt on the balance between the ET-1 and PGI2 system may be involved in its mechanisms.

Objective: To explore the intervention effects of Xiaopi Pill (XPW), a compound traditional Chinese herbal medicine, on the development progress of dimethylnitrosamine (DMN)-induced liver fibrosis in rats., Methods: Liver fibrosis model was established by intraperitoneal injection of 0.5% DMN 2 mL/kg thrice a week for 4 weeks. Rats were divided into control group given saline and treatment group given XPW during the 3rd week of DMN injections. Rats were sacrificed at the end of the experiment, and then liver histological changes, liver function and mRNA expression of the liver fibrosis-associated markers were observed., Results: (1) At the end of the 2nd and 4th weeks of DMN injection, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) increased significantly in rats (P<0.01 or P<0.05); content of total bilirubin (TBil) increased significantly compared with the normal group until the end of the 4th week (P<0.05); compared with the model group after 4 weeks of DMN injection, the serum levels of ALT, AST, ALP and TBil were decreased remarkably in the XPW-treated group (P<0.01 or P<0.05). (2) The hepatic inflammation and collagen deposition in hepatic tissues increased by different degrees in experimental rats. Parts of pathological changes in the rat liver were found at the end of the 4th week, including a complete round structure of false flocculus round, meantime, the hydroxyproline content of hepatic tissue was increased significantly at the end of the 2nd and 4th weeks (P<0.05). Compared with the 4-week model group, the hepatic inflammation, collagen deposition and hydroxyproline content in hepatic tissues were alleviated dramatically (P<0.05). (3) Compared with the normal and 2nd week groups, protein expression of alpha-smooth muscle actin (α-SMA) was gradually increased, and that of the 4th week group were aggrandized significantly (P<0.01). Compared with the normal group, the mRNA expression of α-SMA, transforming growth factor-β1 (TGF-β1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and heme oxygenase-1 (HO-1) was gradually increased. Further changes in above-mentioned abnormalities were found in the model rats at the end of the 4th week (P<0.01); while compared to the 4th week group, protein and mRNA levels of α-SMA and mRNA levels of TGF-β1, TIMP-1, and HO-1 were decreased significantly in the XPW group (P<0.01 or P<0.05)., Conclusion: Progressive DMN-induced liver fibrosis in rats can be suppressed by XPW; the mechanism may be associated with inhibition of the activated hepatic stellate cells.

Objective: To investigate the therapeutic effect of Corbrin shugan capsule for treatment of alcoholic hepatic fibrosis in rats., Methods: The rat model of alcoholic hepatic fibrosis was induced by intragastric administration of alcohol repeatedly. The serum procollagen III (PC III), laminin (LN) and tissue inhibitors of metalloproteinase-1 (TIMP-1) levels were measured with ELISA, and the content of hydroxyproline (Hyp) in liver tissue were determined with colorimetric method. Collagen deposition in liver tissue was observed with Masson's staining, and the fibrosis area was measured with digital medical image analysis system (Motic Med 6.0)., Results: Compared with the model control group, the serum TIMP-1 and LN levels and hepatic fibrosis area in liver tissue significantly decreased in Corbrin shugan capsule groups with doses of 0.09,0.27 and 0.45 g*kg(-1), and the serum PC III and the Hyp contents in liver tissue also decreased of Corbrin shugan capsule groups with doses of 0.27 and 0.45g*kg(-1)., Conclusion: Corbrin shugan capsule can decrease serum PC III, TIMP-1 and LN levels and Hyp levels in liver tissue and hepatic fibrosis area in rats, indicating it may have therapeutic effect on alcoholic hepatic fibrosis.

The study is to observe the effect of racemic TJ0711 on blood pressure and heart rate as well as protection of cardiovascular system of renal hypertensive rats after long-term administration. The renal hypertensive models were established by the two-kidney, one-clip (2K1C) method in Wistar rats. Four weeks later, assigned the rats whose SBP had increased at least 4 kPa randomly into 5 groups: racemic TJ0711 10, 20 and 40 mg x kg(-1) groups, carvedilol control group, model group and sham group (n=10), ig administration once daily. The changes of BP (blood press) and HR (heart rate) before and after administration were measured by tail-cuff method weekly. Plasma samples of all animals were taken in 6-8 weeks, and plasma MDA as well as renin, angiotensin II (Ang II) and endothelin-1 (ET-1) levels were measured. Left ventricle was cut off after 9 weeks, and left ventricular weight index (LVWI) and hydroxyproline were measured. The significant decrease of the BP of TJ0711 40 mg x kg(-1) group was observed after TJ0711 ig administration for 4 weeks, and this effect remained till the end of the study. In 8th week, the systolic blood pressure values were: TJ0711 40 mg x kg(-1) group 18.93 +/- 1.82 kPa (vs 21.30 +/- 2.30 kPa, P < 0.05); 20 mg x kg(-1) group 20.68 +/- 3.29 kPa (vs 22.19 +/- 2.88 kPa). The plasma MDA level of all treated groups was significantly lower than that of model group, so were the plasma renin, Ang II and ET-1 levels (P < 0.05). LVWI and hydroxyproline content of myocardial tissue decreased to some extent, but was not significant as compared with that of model group. The study showed that TJ0711 repeated dosing could reduce BP level beginning from drug administration; besides block adrenal alpha and beta receptors to play an antihypertensive role. The sustained antihypertensive effect also related to reduce plasma vasoconstrictor substances and oxidation product MDA. These effects benefited cardiovascular protection.

This study is to determine the preventive effect and mechanism of targeting IL-17A on pulmonary inflammation and fibrosis after acute lung injury. Mice were treated with anti-IL-17A antibody on the day 7 and sacrificed on the day 14 after bleomycin lung injury. The pulmonary inflammatory status and the deposition of collagen were measured by HE and Sirius stains staining. The contents of hydroxyproline and collagen were measured by using commercial kits. The survival rate of mice was calculated by Kaplan-Meier methods. The inflammatory cytokines in bronchoalveolar lavage fluid were measured by ELISA and the expressions of inflammation-related molecules were detected by Western blotting assay. Targeting of IL-17A could prevent the development of lung inflammation, decrease collagen deposition and the contents of hydroxyproline, and protect against the development of pulmonary fibrosis, which together led to an increase in the animal survival. Moreover, blocking IL-17A decreased the expression ofpro-fibrotic cytokines such as IL-17A, TGF-beta1 and IL-13; increased the expression of anti-fibrotic or anti-inflammatory factors such as IFN-gamma, COX-2, 5-LOX, 15-LOX. Indeed, IL-17A antagonism suppressed the activation of pro-inflammatory p65NF-kappaB but enhanced the activation of pro-resolving p50NF-kappaB. In conclusion, that blockade of IL-17A prevents the development of pulmonary fibrosis from acute lung injury, is because blocking IL-17A may prevent acute inflammation converting to chronic inflammation.

Objective: To observe the effect of traditional Chinese medicine formulas clearing away heat and promoting blood circulation-Biejiayinzi (BJYZ), Gexiazhuyu Tang (GXZYT) and Fugan Wan (FGW) on liver fibrosis in CCl4 mice by screening and analyzing formula-syndrome database of kidney and liver fibrosis based on the principle of formula-syndrome, compared with pivot-harmonizing decoction., Method: Ten-week-old male C57BL/6 mice, with the weight of (20 +/- 3) g, were randomly divided into 6 groups: the normal group, the model group, the BJYZ group, the GXZY group, the FGW group and the XST group. Except the normal group, other groups were abdominally injected with 10% CCl4 olive oil solution a dose of 2 mL x kg(-1) body weight for four weeks, three times each week. Meanwhile, the latter four groups were administered with extracts of BJYZ, GXZYT, FGW and XST, respectively, once every day, concomitantly continued CCl4 administration. The normal and the model groups were given the same volume of deionized water. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (gamma-GT), Alb and TBil were detected by chemiluminescence. The hydroxyproline (HYP) content was detected by acid hydrolysis method. The hepatic collagen deposition was evaluated with Sirius red staining., Result: Compared with the normal group, the model group recorded notable decrease in weight and increase in the ratio of liver weight and body weight and the ratio of spleen weight and body weight, with obvious fatty degeneration and inflammatory necrosis in liver cells. Collagen fiber deposition was so notable to form fibrous septums and pseudolobules. The levels of serum ALT, AST, TBil, gamma-GT, the HYP content in liver tissue and the deposition of hepatic collagen were significantly increased in the model group. Compared with model group, Serum AST were significantly decreased in BJYZ group as gamma-GT decreased in the GXZYT group, without notable decrease in degeneration and inflammatory necrosis in liver cells and collagen deposition. The GXZYT group showed significant decrease in gamma-GT, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The ratio of liver weight and body weight, AST, gamma-GT and HYP content were significantly decreased in the FGW group, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The XST group showed decrease in the ratio of liver weight and body weight, with no obvious change in inflammation and fibrosis of hepatic tissue., Conclusion: FGW shows the best effect of prevention and treatment of liver fibrosis in CCl4 mice.

Objective: To observe the effects of blueberry on rat with hepatic fibrosis and ultrastructural. of hepatocytes., Methods: Sixty (60) healthy Wistar rats were randomly divided into six groups: normal control group (group A), hepatic fibrosis model group (group B), blueberry at low, middle and high concentration groups (group C, D, E), Fu-Fang-Bie-Jia-Ruan-Gan tablet group (group F). The hepatic fibrosis model of rat was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in groups C-F were respectively perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet for 12 weeks except for the normal control group which accepted saline alone. Upon terminal sacrifices of all rats at the end of the twelve weeks. Pathology of hepatic tissue was evaluated by hematoxylin-eosin (HE), Masson staining and transmission electron microscope. Liver index were measured. Levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were examined. Activities of superoxide dismutase (SOD), the contents of malondialdehyde (MDA), hydroxyproline (Hyp) and reduced glutathione (GSH) in liver homogenates were determined., Results: Refer to the serum levels of ALT and AST, there is no significant difference existed in all groups (P > 0.05). Activities of SOD, the contents of GSH in liver homogenates of group D and E were significantly higher than those of group B(P < 0.05), while liver index, the contents of Hyp, MDA in liver homogenates were significantly lower than those of group B (P < 0.05). Compared with the group B, the pathological stages of hepatic fibrosis in group D and E were significantly reduced (P < 0.05), the expression of collagen were decreased, ultrastructural alterations were markedly improved., Conclusion: Blueberry may have certain preventional protective effects on porcine serum induced rat hepatic fibrosis and exhibit certain protective effects on organelles of hepatocytes (such as mitochondria), especially in middle and high does of blueberry. Its mechanism is probably related to increase the ability of antioxidant stress by increasing SOD activity and GSH content and reducing MDA content.

Objective: To study the effect of gypenosides on DMN-induced liver fibrosis in rats., Method: A rat liver fibrosis model was established by injecting DMN intraperitoneally. Four weeks later, model rats were randomly devided into three groups: the model group, the gypenosides treated group (200 mg x kg(-1)) and the colchicine treated group (0.1 mg x kg(-1)), with 10 specimens for each group. After a 2-week treatment, following parameters were observed: (1) last body weight, weight ratio between liver and spleen; (2) content of liver hydroxyproline (Hyp); (3) activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (gamma-GT), content of albumin (Alb) and total bilirubin( TBiL) in serum; (4) liver pathology (Sirius red staining and HE staining); (5) activity of liver superoxide dismutase (SOD), glutathione reduced (GSH), glutathione peroxidase (GSH-Px) and content of liver maleic dialdehyde (MDA)., Result: There were classic liver cirrhosis pathological changes in model groups. Compared with the normal group, liver Hyp content, activity of serum ATL, AST, gamma-GT and content of serum TBiL, MDA of model groups significantly increased; content of serum Alb and liver GSH, activity of liver SOD and GSH-Px decreased significantly in model groups. In comparison with the model group, liver cirrhosis remarkable improved in the gypenosides group, content of liver Hyp reduced significantly (P < 0.01), which was equal to the colchicine group. Compared with the model group, liver function parameters improved markedly in the gypenosides group; liver SOD and GSH-Px activities significantly increased; MDA content reduced significantly (P < 0.05)., Conclusion: Gypenosides shows an effect in treating DMN-induced liver fibrosis in rats.

Objective: To investigate hydroxyproline tolerance of Chrysanthemum morifolium plantlets included " Boju", "Huaiju", "Chuju", "Gongju" and "Hangju",and provide references basis for excellent cultivar and breeding of Ch. morifolium., Method: Plantlets in vitro from five kinds of Ch. morifolium were inoculated on medium added with different concentrations of hydroxyproline. Free proline in leaves from plantlets was determined, then the damage index and survival rate were compared., Result and Conclusion: The results showed that hydroxyproline tolerance of " Boju" and "Huaiju" were superior, the survival rates and free proline of them were higher, but the damage index was inferior. The hydroxyproline tolerance of "Hangju" was the worst, and the survival rate was minimum. The survival rate of "Chuju" and "Gongju" was between "Boju" and " Hangju", and the hydroxyproline tolerance of them was also medium.

Objective: To investigate the effects of Leonurus japonicas on myocardial remodeling induced by isoproterenol (ISO) in rats., Methods: The model of rat myocardial remodeling was established by subcutaneous injection of ISO (20,10 and 5 mg/kg for 3 days, subsequently 3 mg/kg for 7 days). Rats were randomly divided into five groups: Control, ISO, ISO + enalapril, ISO + Leonurus 8 g/ kg and ISO + Leonurus 16 g/kg. Recorded the values of LVSP, LVEDP, +/- dp/dt (max), cardiac output ( CO). Weighed the body weight (BW), heart weight (HW) and left ventricular weight (LVW) of rats to calculate the values of HW/BW and LVW/BW. The hydroxyproline contents were investigated by spectrophotometric measurement. The contents of type I collagen and type III collagen and ratio of I/III collagen were assessed by immunohistochemical stain., Results: Leonurus (16 g/kg x day(-1)) could increased the value of LVSP, + dp/dt(max), CO (P < 0.05). Leonurus (8 g/kg) could increased the value of -dp/dt(max), decreased the ratios of HW/BW and LVW/BW (P < 0.05), the contents of hydroxyproline, type I collagen, type III collagen and the ratios of I/III collagen (P < 0.05)., Conclusions: Leonurus at a dosage of 16 g/kg may improve the systolic function; Leonurus at a dosage of 8 g/kg may improve the diastolic function, down-regulate the expression of collagen and normalize the ratio of I/III collagen.

This study is to investigate the therapeutic effect and mechanism of indole-3-carbinol (I3C) on pig serum-induced liver fibrosis of rats. The liver fibrotic model of rats was induced by pig serum. After models were successfully established, rats in the treatment groups were administered with I3C through intraperitoneal injection or curcumin by intragastric administration, daily for 17 days. Hepatic hydroxyproline (Hyp) content was measured. The liver histology and immunohistochemistry with a-smooth muscle actin (alpha-SMA) were assayed. Hepatic stellate cells line, HSC-T6 was incubated with different concentrations of I3C (25, 50, and 100 micromol x L(-1)) for 24 h. The effect of I3C on cell apoptosis was identified by FITC-Annexin V/PI double labeled assay. And the mRNA expressions of Bax and Bcl-2 were measured by real time RT-PCR. The results showed that hepatic content of Hyp decreased by I3C treatment, as compared with the fibrotic model control. Histopathological changes, such as steatosis, necrosis, deposition of collagenous fiber reduced remarkably and the expression of alpha-SMA was significantly down-regulated in the I3C-treated groups (P < 0.01). Apoptosis analysis showed that I3C significantly increased HSC-T6 apoptosis rate and the expressional ratio of Bax to Bcl-2. The results indicated that I3C could effectively cure pig serum-induced liver fibrosis in vivo by inducing HSC apoptosis and promoting ECM degradation.

Aim: To observe the effect of serum concentration on the growth, proliferation and secretion of collagen I, collagen III and hydroxyproline in mouse fibroblasts L929, to optimize serum concentration for cell growth, proliferation and collagen secretion., Methods: L929 cells were divided into 6 groups cultured in the present of 0 mL/L, 200 mL/L, 400 mL/L, 600 mL/L, 800 mL/L and 1 000 mL/L serum respectively. Acridine Orange and HE staining were performed on 2 d, 4 d, 6 d and 8d and sulforhodamine B sulfonyl chloride was used to determine cell proliferation at the same time; Meanwhile, the supernatants of each group ware collected and enzyme immunoassay was performed to detect the protein levels of collagen I, III and hydroxyproline., Results: (1) The cell proliferation was inhibited when serum concentration was very high (100%) or very low (0%). The proliferation rate in the presence of 20% serum was the highest. When serum concentration was higher than 20%( 40%-100%), the cell proliferation decreased. (2) Apoptotic cells were observed in the absence of serum. Vacuolization appeared in the presence of 80% serum, and was enhanced in the case of 100% serum. Cells were morphologically normal when serum concentrations were between 20%-60%. (3)The level of collagen I was low in the case of serum concentrations of 0%, 80% and 100%, with the highest expression in 40% serum. Collagen III in all groups was increased on day 2, followed by a transient decreased and an final increased on day 6-8. The level of hydroxyproline was not changed in all groups., Conclusion: The serum concentrations between 20%-40% is suitable for L929cells growth, proliferation and secret collagen. The extremes serum concentration of too high or too low leads to inhibition of cell proliferation, apoptosis, vacuolization, and impaired secretion of collagens. This study indicates the requirement of optimum serum concentration for self serum injection therapy.

The aim of this study was to evaluate the efficacy of Bletilla striata polysaccharide on diabetes mellitus ulcers. Diabetes mellitus animal model was established by single ip injection of streptozotocin (STZ, 50 mg x kg(-1)) with the criteria of blood glucose > or = 16.7 mmol x L(-1) after 72 h. 4 weeks after STZ injection, each animal received two full thickness incisional wounds (1.8 cm in diameter). The wounds then were divided into B. striata polysaccharide group and PBS group. Wound closure rate, fibroblast (FB) infiltration, hydroxyproline (OHP) content and myeloperoxidase (MPO) levels were examined on day 3, 7, 14, 21 post wound. The treatment of B. striata polysaccharide significantly facilitated diabetes mellitus ulcers healing compared to PBS group. Histological analysis showed that B. striata polysaccharide markedly increased inflammatory cell infiltration in wound area. The herb also strongly evaluation of FB, OHP demonstrated a significantly increased in B. striata polysaccharide group. B. striata polysaccharide group promoted wound closure by means of enhanced inflammatory cell infiltration and re-epithelialization, and the promotion of FB and OHP levels.

To clarify the mechanism underlying the preventive effect of baicalin (Bai) on fibrosis in lung, we investigated the influence of Bai on the up-regulation of connective tissue growth factor (CTGF) in fibrotic lungs. Male Sprague-Dawley (SD) rats were divided into four groups randomly: normal saline (NS)+NS group (a single intratracheal instillation of NS plus i.p. injection of NS), NS+Bai group (intratracheal instillation of NS plus i.p. injection of Bai), bleomycin (BLM)+NS group (intratracheal instillation of BLM plus i.p. injection of NS) and BLM+Bai group (intratracheal instillation of BLM plus i.p. injection of Bai). All the i.p. injections were performed once daily. On day 28 after intratracheal instillation of BLM or NS, the rats were sacrificed for lung tissue sampling. As the index of the severity of pulmonary fibrosis, the content of hydroxyproline in lungs was analyzed by chloramine T method. The expression levels of CTGF mRNA and protein in the lungs were detected by RT-PCR and immunohistochemistry, respectively. The results showed that, compared to the rats in NS+NS group, the rats in BLM+NS group showed increased hydroxyproline content and higher levels of CTGF mRNA and protein expressions (P<0.01), suggesting that BLM had induced fibrosis in lung and up-regulated CTGF expression in the fibrotic lungs. Administration of different dosages of Bai (6, 12.5 and 50 mg/kg per d, for 28 days) into the BLM-treated rats reduced the increased content of hydroxyproline, and ameliorated the up-regulation of CTGF mRNA and protein levels, respectively. These results suggest that Bai could prevent the up-regulation of CTGF expression in fibrotic lungs of rats receiving BLM instillation, which might be one of the mechanisms underlying the preventive effect of Bai on pulmonary fibrosis.

Objective: to investigate whether pirfenidone (PFD) presents the antifibrotic effect in silicosis of rats., Methods: SD rats were randomly divided into five groups: the non-treat group, the normal saline group, the normal saline + PFD group, the SiO2 group, the SiO2 + PFD group. Rats except in the non-treat group were intratracheally instilled with SiO2 (25 mg/ml) or normal saline. The rats in normal saline + PFD group and the SiO2 + PFD group were given PFD (50 mg/kg) orally the next day after instillation and throughout the study. Rats were respectively sacrificed 7, 21, 42 days after instillation. The pathology changes were evaluated by Haematoxylin-eosin (HE), Van Gieson and Foot staining, and the hydroxyproline (HYP) content of pulmonary tissue was determined., Results: compared with the SiO2 group, PFD could relieve the fibrotic changes in the lungs of rats. The fibrotic degree in silicotic lesions of lungs was lower in the SiO2 + PFD group than that of SiO2 group. The HYP content in the lungs of the SiO2 + PFD group [(0.75 ± 0.12) mg/g] was significantly lower than that of the SiO2 group [(1.19 ± 0.17) mg/g] at 42 days after instillation (P < 0.05)., Conclusion: these data support that PFD has an antifibrotic effect against SiO2 induced lung fibrosis in rats, Which appears to be changing collagen accumulation and inhibiting pulmonary fibrosis.

Objective: To observe the therapeutic effect and mechanism of penehyclidine hydrochloride on paraquat-induced acute lung injury., Methods: 80 healthy adult male Wistar rats were randomly assigned into control groups (10 rats), 100 mg/kg PQ group (10 rats), 100 mg/kg PQ plus 33 µg/kg penehyclidine hydrochloride treatment group (30 rats), 100 mg/kg PQ plus 66 µg/kg penehyclidine hydrochloride treatment group (30 rats). The two treatment groups were executed respectively at 36 h, 72 h and 7 d. Lung tissues were used to assess histopathological change by HE staining. The level of MMP-2, caveolin-1 and HYP were detected in the lung homogenate. The serum and BALF contents of ET were measured., Results: Pathology inspection confirmed that the model of acute rat pulmonary injury were duplicated successfully. The level of MMP-2, HYP in lung tissues and the serum and BALF ET contents in PQ group were (1.77 ± 0.40) µg/g, (2.91 ± 0.79) µg/g, (505.23 ± 124.69) µg/ml, (640.38 ± 136.60) µg/ml. The level of those was higher than that in control group [(0.95 ± 0.66) µg/g, (1.48 ± 0.69) µg/g, (95.48 ± 46.01) µg/ml, (200.40 ± 88.39) µg/ml, P < 0.05]; The above-mentioned index in two treatment groups was lower than that in PQ group (P < 0.05). The caveolin-1 content [(1.77 ± 0.82) µg/g] in PQ group was lower than that in control group [(5.39 ± 1.68) µg/g, P < 0.05]. The level of caveolin-1 in two treatment groups was higher than that in PQ group (P < 0.05)., Conclusion: Penehyclidine hydrochloride can decrease the level of MMP-2, HYP in lung tissues and the ET in serum and BALF, increase that of caveolin-1 and lessen the damage induced by paraquat.

Objective: To explore the stimulation of human hepatic stellate cells by Cytochrome P4502E1-mediated oxidative stress., Methods: HepG2-line was transfected with human CYP2E1 plasmid (HepG2/CYP2E1) and empty plasmid (HepG2/PCI) respectively. The CYP2E1 expression was evaluated with RT-PCR and Western blot. MDA was measured in culture medium of HepG2 cell lines. LX2 was co-incubated with HepG2/CYP2E1, HepG2/PCI and HepG2 respectively. The level of hydroxyproline in culture medium was examined in 48 hours and the cells were lysated and total RNA and protein were extracted. COL-1 and MMP2 mRNA levels were detected by RT-PCR and analyzed semi-quantitatively. PICP proteins were measured by ELISA. Zymography was performed to investigate MMP2 enzymatic activities., Results: (1) MDA from the HepG2 which (HepG2/CYP2E1)express human CYP2E1 (6.51+/-0.25) was significantly higher than that from the HepG2 which do not (HepG2/PCI) express human CYP2E1 (3.07+/-0.29) and HepG2 alone (2.57+/-0.29). (F=22.66, all P<0.01). (2) After co-incubated for 48 hours,the level of hydroxyproline in culture medium (35.24+/-3.52) excreted from CYP2E1/LX2 could significantly increase (F=58.89, P is less than 0.01). PICP protein (540.01+/-11.38) excreted from CYP2E1/LX2 was significantly increased (F=124.97, P<0.01). Zymography showed MMP2 gene expression and enzymatic activities of MMP2 had no difference among the groups (F=0.29, P>0.05) (F=0.33, P>0.05)., Conclusions: CYP2E1 derived oxidative stress mediated stimulation of collagen I synthesis by hepatic stellate cells. Hydroxyproline excreted by LX2 was increased by CYP2E1. COL-1mRNA had no difference among the groups (F=0.73, P>0.05).

The purpose of this study is to evaluate the effect of oligomeric grape seed proanthocyanidins (GSP) on isoproterenol (ISO)-induced cardiac remodeling in rats. ISO was given subcutaneously (5 mg x kg(-1), sc, 7 days) to induce cardiac remodeling in rats. Therapeutic groups were given GSP (50, 100, and 150 mg x kg(-1)) after ISO treatment. After 2 weeks intervention, heart rate (HR), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), rate of rise of left ventricular pressure (+/- dp/dt(max)) were examined. The myocardial hypertrophy index was expressed as heart weight/body weight (HW/BW) and left ventricle weight/body weight (LVW/BW), the histological changes were investigated by HE and Van Gieson stain. SOD activity and MDA content in serum, contents of hydroxyproline (Hyp) in the left ventricular tissue were assayed by xanthinoxidase method, thiobarbituric acid (TBA) method and alkaline hydrolysis method, respectively. After the onset of ISO-treatment, GSP therapy potently improved cardiac function, inhibited myocardial hypertrophy, improved cardiac pathology change, decreased the myocardial cross-section area (CSA), collagen volume fraction (CVF) and perivascular circumferential collagen area (PVCA), reduced the content of Hyp in the left ventricular tissue, inhibited the decrease of SOD activity and increase of MDA content in serum. GSP possess protective effect against ISO induced cardiac remodeling in rats, this may be related to reducing the oxidative stress and improving antioxidant capacity.

Objective: To investigate the effect of Aloe polysaccharide on proliferation and hyaluronic acid and hydroxyproline secretion of human fibroblasts in vitro., Methods: The fibroblasts were treated with different doses of polysaccharide (0, 25, 50, 100, 200, 400 mg/L). Subsequently, cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell cycle by flow cytometry, evaluation of the Aloe polysaccharide toxic effect by acridine orange-ethidium bromide staining, evaluation of the cell injury by lactate dehydrogenase (LDH) assay, and the collagen synthesis by (3)H-proline incorporation. In addition, hyaluronic acid and hydroxyproline levels in the supernatants of cultured fibroblasts were measured by enzyme-linked immunosorbent assay., Results: The proliferation of fibroblasts was induced with polysaccharide in a dose-dependent manner, reaching its highest level on 5th day. Meanwhile, the percentage of cells at phase G(0)/G(1) was decreased, while that at phases G(2)/M and S was increased significantly in Aloe polysaccharide-treated groups as compared with those in the control group (P<0.05). Additionally, the apoptosis of the fibroblasts showed no differences among all groups. The collagen synthesis was increased and cell injury decreased in polysaccharide-treated groups as compared with those in control group (P<0.05), while the levels of hyaluronic acid and hydroxyproline in the supernatants of fibroblasts treated with polysaccharide were significantly higher than those in the control group (P<0.05)., Conclusion: The Aloe polysaccharide promotes both the proliferation of fibroblasts and the production of hyaluronic acid and hydroxyproline in fibroblasts. This indicates that the Aloe polysaccharide may play an important role in the extracellular matrix remodeling during wound healing.

Objective: To explore therapeutic effect of Haobieyangyinruanjianfang (HBYYRJ) on mouse liver fibrosis by schistosomiasis., Methods: Mice except for normal control were infected with Japanese schistosome cercarias, after 12 weeks, infected mice were divided into 7 groups: low HBYYRJ group, middle HBYYRJ group, high HBYYRJ group, Fufangbiejiaruangan tablet (FFBJRG) group, colchicine group, 3 months infection group and 6 months infection group. Hepatic fibrosis was found in 3 months infection group. Liver hydroxyproline (Hyp) was determined, matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) were detected with gelatin zymography, serum hyaluronic acid (HA) and precollagen III (PC-III) were detected using RIA., Results: HBYYRJ obviously reduced hepatic fibrosis (probability value less than 0.01). Collagen and HA in 3 months infection group and 6 months infection group were higher than that in normal group (probability value less than 0.01), collagen in high and middle HBYYRJ groups and HA in middle and low HBYYRJ groups were lower than that in 6 months infection group (P less than 0.01, probability value less than 0.05). The expression of MMP-9 and MMP-2 in 3 months infection group and 6 months infection group was higher than that in normal group (probability value less than 0.01), The expression of MMP-9 in three HBYYRJ groups and the expression of MMP-2 in high HBYYRJ group were lower than that in 6 months infection group (probability value less than 0.05)., Conclusion: HBYYRJ can reduce liver fibrosis caused by schistosomiasis.

Objective: To investigate the effects of anluohuaqianwan on experimental hepatic fibrosis induced by dimethyl nitrosamine (DMN) in rats., Methods: 36 male SD rats were randomly dividied into three groups: model group, normal group, anluohuaqianwan group. The rats in the three groups were treated with DMN daily for 4 weeks. The liver function was detected using auto biochemistry analyzer, the serum HA, LN, IV-C, PIIIP were detected by immunoradiometry, the histopathology was observed in the left liver lobe after HE staining, the expression of matrix metalloproteinase-2 (MMP-2) in liver tissue were detected by immunohistochemistry., Results: The serum levels of ALT, AST, ALP, TP, ALB and the contents of HA, LN, IV-C in model group were significantly increased compared to these in the normal group (P less than 0.01). The serum levels of ALT, AST and the contents of HA in anluohuaqianwan group were significantly lower than those in the model group (P less than 0.01). The liver MMP-2 in the model group was significantly increased compared to that in the normal group (P less than 0.05). The expression of MMP-2 in liver tissue of model group was lower than that in the anluohuaqianwan group (P less than 0.05)., Conclusion: Anluohuaqianwan can inhibit liver fibrosis in rats induced by DMN.

Objective: To investigate the effect of urokinase on hepatic fibrogenesis in rats., Methods: Hepatic fibrosis was induced in rats by complex pathogenic factors including subcutaneous injections of carbon tetrachloride, alcohol and cholesterol feeding. Animals were randomly divided into 3 groups: normal control group, hepatic fibrosis group (complex pathogenic factors for 6 weeks), UK prevention group (complex pathogenic factors+UK for 6 weeks). The animals were sacrificed at the end of week 6. The expression of alpha-SMA, uPA, PAI-1, TGFb1, TIMP-1, collagen type I and type III proteins in hepatic fibrosis tissue was detected by immunohistochemistry, the expression of PAI-1 and TGFb1 mRNA in the hepatic fibrosis tissue was quantified by real time RT-PCR. The serum levels of hyaluronicacid (HA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin (TBil) and the content of liver hydroxyproline (Hyp) were detected using ELISA kits., Results: The serum ALT, AST, TBil, HA and the content of liver Hyp were (46.66+/-6.30) U/L, (126.26+/-31.65) U/L, (31.11+/-4.20) micromol/L, (109.70+/-18.81) microg/L and (0.98+/-0.09) mg/(g liver), respectively, in UK prevention group, which were significantly lower than those [(101.57+/-11.97) U/L, (205.89+/-56.26) U/L, (67.75+/-2.75) micromol/L, (184.43+/-32.36) microg/L and (1.65+/-0.16) mg/(g liver), respectively] in hepatic fibrosis group (q = 3.3801-20.0061, P < 0.01). The levels of a-SMA, collagen type I, type III, TIMP-1, PAI-1, TGFb1 proteins were (299.27+/-37.36), (210.05+/-27.17), (192.94+/-24.48), (213.70+/-32.21), (204.25+/-17.92), (205.97+/-23.81), respectively, in UK prevention group, which were significantly lower than those [(418.83+/-30.21), (323.77+/-21.53), (302.37+/-31.43), (376.63+/-25.19), (313.53+/-26.67) and (327.42+/-36.75), respectively] in hepatic fibrosis group. The level of uPA protein was increased, and the expression of PAI-1, TGFb1 mRNA in hepatic fibrosis tissue was decreased in UK prevention group., Conclusion: In the early stage of hepatic fibrogenesis, urokinase can attenuate the progression of rat hepatic fibrosis via upregulation of uPA, downregulation of TGFb1, and inhibition of HSC activation.

Objective: To study the effects of artemether and dihydroarteannuin on the mouse model of scleroderma., Method: Sixty mice were randomly divided into 8 groups: PBS control group, model group, menstruum group (20% Tween-80, 0.4%CMC-Na), positive medicine group (penicillamine 200 mg kg(-1)), low-dose artemether group (5 mg kg(-1)), high-dose artemether group (20 mg kg(-1)), low-dose dihydroarteannuin group (5 mg kg(-1)), high-dose dihydroarteannuin group (25 mg kg(-1)). We have established a mouse model for scleroderma in Balb/c mice by subcutaneous injections of bleomycin 0.1 mL per day (200 mg L(-1) BLM) for 3 weeks. Meanwhile, the administration lasted for 4 weeks. The back skin was removed in the next day after the final administration. Treated skins and lungs were harvested and analyzed for histological sclerosis. The thickness of the skin and fibrosis degree of derma were observed and made an analysis of the contents of collagen and hydroxyproline., Result: Compared with the model groups, the high-dose groups markedly inhibited the thickness of derma (P<0.001), furthermore, the contents of collagen and hydroxyproline in the skin were also significantly reduced (P<0.05). Other groups of mice showed improvement on scleroderma., Conclusion: Our results suggest that administration of artemether or dihydroarteannuin may be an effective approach in preventing systemic sclerosis.

Aim: To investigate whether Smad6 and Smad7 can regulate TGF-beta-induced epithelial-mesenchymal transition in human renal proximal tubule epithelial cells., Methods: Two recombinant adeno-associated viruses (AAV) expressing Smad6 and Smad7 genes were produced without helper virus and then they were delivered into human renal proximal tubule epithelial cells (HKCs). The cells were randomly divided into normal controls, TGF-beta1-treated group, Smad7-infected control, LacZ-infected control, TGF-beta1+Smad7 group or TGF-beta1+Smad6 group, and TGF-beta1 + LacZ group. 10 microg/L of TGF-beta1 was added into the cell culture at the time of 15 min, 30 min, 60 min, and 120 min and the third day. The levels of phospho-Smad2, alpha-smooth muscle actin (alpha-SMA) and E-cadherin proteins were measured by Western blot. The concentration of hydroxyproline excreted into the culture supernatant was determinated by ELISA. The morphological changes of the cells detected by inverted microscope., Results: Compared with the cells in absence of TGF-beta1, the expression level of P-Smad2 in HKCs increased at 15 min, 30 min, 60 min, and 120 min with the TGF-beta1 stimulation. It reached peak at 30 min. TGF-beta1 treatment for 72 hours resulted in significant up-regulation of alpha-SMA protein expression and hydroxyproline secretion, but a marked decrease in E-cadherin expression in the culture of HKCs. 10 microg/L of TGF-beta1 treatment for 72 hours also resulted in marked alteration in cell morphology. The cells lost their regular cuboidal appearance, and become elongated and spindle shaped. In the Smad7-infected cells, the overexpression of Smad7 resulted in a marked decrease of alpha-SMA protein and hydroxyproline synthesis, but a increase of E-cadherin protein, as well as the retainment of epithelial phenotypic appearance. These effects were associated with the inhibition of TGF-induced Smad2 activation, whereas the overexpression of Smad6 had no effect on the TGF-beta-induced changes in HKCs., Conclusion: The overexpression of Smad7 instead of Smad6 can efficiently inhibit TGF-beta-induced epithelial-mesenchymal transition by blocking Smad2 activation in human renal proximal tubule epithelial cells.

Objective: To determine the effects of curcumin on bleomycin (BLM)-induced pulmonary fibrosis in rats., Method: One hundred and forty-four male Sprague-Dawley rats were randomized into 6 groups (24 rats in each group, model group, sham group, prednisone group (0.56 mg x kg(-1) x d(-1)), curcumin with low dose 5 mg group, curcumin with middle dose group 10 mg and curcumin with high dose group 20 mg per 100 g of body weight). Rats in all groups except in sham group were injected with BLM intratracheally. Curcumin with different doses were given by gavage one time everyday for 7, 14 and 28 days. Prednisone were given to rats in prednisone group, po, serving as the positive treatment group. On the 7th, 14th, 28th day, the lung functions (inspiratory resistance, maximal volutary ventilation, forced vital capacity, Fev 0.2/FVC, peak expiratory flow) were determinated in experimental rats, respectively, and the concentrations of hydroxyproline in lung homogenates of each rat were assayed., Result: Administration of curcumin in different doses improved lung functions of BLM-induced fibrotic rats in the all experimental days; and it decreased the concentration of hydroxyproline in lung homogenates compared with those levels in model control group; and it also lessened the hyperplasia of BLM-induced pulmonary fibrosis in rats., Conclusion: Administration of curcumin can suppress BLM induced pulmonary fibrosis indicated by improved respiratory function, as well as companied with low content of hydroxyproline in lung tissue of rats.

Objective: To investigate the effects of carvedilol and metoprolol on cardiac fibrosis in rats with experimental myocardial infarction (MI)., Methods: MI was induced in male Sprague-Dawley rats by ligating the left coronary artery. Rats randomly received saline, carvedilol (10 mg.kg(-1).d(-1)) or metoprolol (20 mg.kg(-1).d(-1)) beginning at 4 weeks post MI for 8 weeks per gavage. Sham-operated rats serve as control. Collagen perivascular circumferential collagen area (PCVA), peri-coronary circumferential collagen area (VLCA) and interstitial collagen volume fraction (ICVF) as well as myocardial hydroxyproline content were determined after hemodynamic measurements at the study end., Results: LVEDP were significantly lower and +/- dp/dt significantly higher in carvedilol and metoprolol treated MI rats than that in saline treated MI rats. Myocardial hydroxyproline, PCVA/VLCA ratio and ICVF were significantly reduced in metoprolol, more significantly reduced in carvedilol treated MI rats compared to saline treated MI rats (all P < 0.05)., Conclusion: Metoprolol and carvedilol could decrease the concentration of hydroxyproline and ICVF in MI rats.

Objective: To study the prevention & treatment effect of cordyceps sinensi on bleomycin-induced pulmonary fibrosis (PF) in mice., Method: Rats were divided into 4 groups. In BM group (model), pulmonary interstitial fibrosis model was established through nasal dripping of 5 mg x kg(-1) bleomycin. In PCT group (prevention & treatment with cordyceps sinensi), cordyceps sinensi was orally administered 48 h before bleomycin dripping. In CT group (treatment with cordyceps sinensi), cordyceps sinensi was orally administered at 14 d after bleomycin dripping, and NC group (normal mice, control). At 28 d, bronchoalveolar lavage fluid (BALF) and lung tissue were collected, right lung received HE staining and masson staining, the change in cell micro-structure was observed under electron microscope, the hydroxyproline (HYP) content in lung tissue of left lung was detected through acidolysis method, and TGF-beta1, IFN-gamma and IL-4 concentration in BALF was detected through ELISA., Result: Compared with those in BM group, mouse lung coefficient decreased significantly (P < 0.05), alveolitis and fibrosis degree improved significantly in PCT and CT groups (P < 0.05), HYP content in lung decreased (P < 0.05). TGF-beta1 and 1L-4 content in BALF decreased significantly in each drug intervention group, IFN-gamma content increased significantly (P < 0.05)., Conclusion: The cordyceps sinensi can alleviate bleomycin-induced alveolitis and fibrosis degree in a certain degree. The mechanism of the alleviation is possibly due to its inhibition of TGF-beta1 expression and its regulation of imbalance of the type I and II cytokine in lung tissue.

Objective: To explore therapeutic effect of glutathione (GSH) on unilateral ureteral obstruction (UUO) induced renal interstitial fibrosis and its mechanism in rat., Methods: Thirty-six adult healthy Wistar rats were randomly divided into 3 groups (each n=12): sham operation group, UUO group and GSH group. Rats in UUO group and GSH group underwent left unilateral ureteral ligation as described previously. Rats in sham group had their ureters manipulated but not ligated. In GSH group, GSH was injected intraperitoneally (100 mg.kg(-1).d(-1)) in different doses based on the animal's body weight, one day before UUO and then for consecutive 10 days after UUO. Meantime, same volume of physiological saline was given in sham operation and UUO groups as GSH group. Animals were sacrificed at 10 days after surgery. The pathological changes in obstructed kidney tissue were observed with hematoxylin and eosin (HE) and Masson stains. The contents of hydroxyproline (Hyp) and total anti-oxygen capability (T-AOC) were measured by chemical colorimetry method, the activity of total superoxide dismutase (T-SOD) was assayed by a modified xanthine/xanthine oxidase method, and the content of malondialdehyde (MDA) was determined by thiobarbituric acid method., Results: Ten days after UUO, swelling and atrophy in renal parenchyma of obstructed kidneys were clearly observed. Fibrous material and monocyte infiltration were increased in the interstitial space. Furthermore, thickening of interstitial space of the tubular basement membrane and widening of the interstitial space of the renal cortex were noted. Hypertrophy or atrophy of juxtaglomerular tubules were also observed. There were cellular or albumin casts in a part of tubules. Interstitial fibrosis was observed in obstructed kidneys 10 days after UUO. The data indicated that ureteral obstruction significantly increased the contents of Hyp and MDA, but decreased the content of T-AOC and T-SOD activity, as compared with sham operation group [Hyp: (0.524+/-0.132) microg/mg, T-AOC: (1.48+/-0.21) U/mg, T-SOD: (12.77+/-0.76) U/mg, MDA: (2.65+/-0.32) nmol/mg, all P<0.01]. Compared with UUO group, pathological changes were milder and the contents of Hyp [(1.598+/- 0.252) microg/mg vs. (1.027+/-0.196) microg/mg, P<0.05] and MDA [(4.58+/-0.59) nmol/mg vs. (3.26+/- 0.34) nmol/mg, P<0.05] were significantly decreased in kidney of GSH group, meanwhile the content of T-AOC was increased [(0.67+/-0.19) U/mg vs. (0.94+/-0.17) U/mg, P<0.05], but the content of T-SOD did not show any change [(9.39+/-0.87) U/mg vs. (9.41+/-0.93) U/mg, P>0.05]., Conclusion: Reduced glutathione treatment attenuates UUO-induced renal interstitial fibrosis via decreasing content of Hyp in UUO kidney and preventing oxidation stress injury.

Objective: To investigate the dynamic expression of placenta growth factor (PLGF) in the lungs with paraquat (PQ)-induced pulmonary fibrosis., Methods: Forty-two adult healthy female Sprague-Dawley (SD) rats were randomly divided into two groups: the control group and the PQ group. Each group was divided into three subgroups, seven animals each. The rats in PQ group were treated intragastrically (ig) with PQ (40 mg/kg) and the rats in control group were treated with the same volume of saline at the beginning of the experiment. The animals of model and control group were sacrificed and lungs were harvested on the 7(th), 14(th) and 28th days respectively. A semiquantitative assay of histological examination and hydroxyproline in lung tissues were used to determine the severity of alveolitis and fibrosis. RT-PCR and immunohistochemistry were used to detect the mRNA and protein expression of PLGF., Results: Hydroxyproline contents in lung tissue were significantly increased after PQ administration. Inflammatory cell infiltration and fibrotic scores were more prominent in the model group compared to the control group. Further study showed that PLGF mRNA on day 7, 14 and 28 (1.28 +/- 0.29, 0.80 +/- 0.07, 0.65 +/- 0.13) and positive index of protein expression (2.27 +/- 0.34, 1.78 +/- 0.41, 1.25 +/- 0.69) in the PQ group were all upregulated as compared with those of the control group., Conclusion: The PLGF expression in the lung tissue in rats with paraquat-induced pulmonary fibrosis is upregulated.

Objective: To study anti-atherosclerotic mechanisms of divided functional recipes of Dahuang Zhechong pill (DHZCP) in rabbits., Method: The atherosclerotic rabbit model was established by high fat feeding combined with immune endothelial injury. Male New Zealand rabbits were divided into 9 groups: normal control group, model control group, Danshen positive control group, and 6 DHZCP-divided groups including divided functional recipes No. 1, 2, 3 with low and high doses for each divided recipe. After intragastric administration for 60 days, blood lipids and serum MDA and NO levels and SOD activity and plasma ET concentration, and contents of hydroxyproline and proteins in the vascular wall were determined., Result: Compared with the model group, the level of blood lipids did not significantly change, serum MDA and ET levels, and the contents of hydroxyproline and proteins in the vascular wall significantly decreased (P < 0.05), and SOD activity and NO level increased in the divided functional recipes (all P < 0.05)., Conclusion: The divided functional recipes of DHZCP can inhibit development of atherosclerosis via a non-lowering lipid mechanisms, including anti-peroxidation of lipids, protection of endothelial function, and decrease of formation of extracellular matrix by reducing synthesis of collage and protein on the vascular wall. Among them, the divided functional recipe No. 1 exhibits the most obvious effect.

Objective: To study the mechanism of Fuzheng Huayu (FZHY) recipe against pulmonary fibrosis relating to MMP-2 activity and type IV collagen expression at lung tissue., Method: The pulmonary fibrosis model was induced by intratracheal instillation with bleomycin once in rats. The models were divided into 3 groups: model control, FZHY recipe treated, and methylprednisolone (Solu-Medrol) treated group, each group was of 14 model rats. Normal control group with 10 rats was intoxicated with the same amount of saline. From the second day of intoxication, FZHY recipe treated group orally took FZHY recipe at the dosage of 4.6 g x kg(-1) rat wt, methylprednisolone treated group received intraperitoneal injection with 15 mg x kg(-1) rat wt of methylprednisolone, while model and normal controls took the same volume of saline, 1 time each day and lasting for 4 weeks. Lung and body weights were weighed and the lung/body ratio was calculated. Collagens deposition was check with Masson stain, and lung hydroxyproline (Hyp) content was assayed with Jamall's method. Protein expressions of MMP-2/9 and type IV collagen at lung tissue were analyzed with Western blot and of MMP-2/9 activities by gelatin zymography., Result: Compared to normal rats, the model control rats had a high lung/body ratio, remarkable collagen deposition, increased Hyp content and the expressions of type IV collagen, MMP-2 and MMP-9 protein at lung tissue, increased MMP-2 activity, in particular active MMP-2 activity, but decreased MMP-9 activity. Compared to model control, FZHY recipe and methylprednisolone obviously attenuated pulmonary collagen deposition, decreased lung/body ratio and Hyp content, downregulated MMP-2 protein expression and activity, in particular active MMP-2 activity, and FZHU recipe had some better actions than methylprednisolone on lung/body ratio, type IV collagen expression and active MMP-2 activity. But both drug groups had no influence on MMP-9 protein expression and activity., Conclusion: FZHY recipe has a good action against experimental pulmonary fibrosis and its mechanisms are associated with the inhibition of MMP-2 protein and activity, and with the inhibition of over expression of type IV collagen at lung tissue.

Objective: To study the preventive and therapeutic effect of Huaxian Decoction (HXD) on bleomycin-induced pulmonary fibrosis in mice by taking treatment of ligustrazin and prednisone as controls, for investigate the clinical value and possible mechanism in treatment of pulmonary fibrosis by traditional Chinese herbs., Methods: A total of 96 male ICR mice, aged 6 to 8 weeks, were randomly divided into 8 groups. Pulmonary fibrosis was induced in the model group by intranasal instillation of bleomycin 5 mg/kg. HXD were given via gastrogavage to the HXD prevention group and treatment group respectively at 48 h before and 14 days after they were modeled with the same method, and the groups of ligustrazin and prednisone, treatment and prevention, were managed the same as for HXD groups, but with ligustazin and prednisone instead of HXD respectively. Bronchoalveolar lavage fluid (BALF) and lung tissue of animals were gathered on day 28. Cellular ultrastructural changes of lung was examined under electron microscope by HE, Masson stain; Content of hydroxyproline (HYP) in lung tissue was assessed by acidolysis method; and concentration of transforming growth factor-beta1 (TGF-beta1) in BALF was assessed by ELISA., Results: Pulmonary coefficient was significantly lower in all the drug intervening groups than that in the model group (P < 0.05), improvement on alveolitis and pulmonary fibrosis were alsonaged the safter intervention, especially in the HXD prevention group (Pinstead < 0.05). HYP content in lung tissue and TGF-beta1 content in BALF lowered at the same time., Conclusion: Early stage application of HXD can alleviate bleomycin-induced alveolitis and pulmonary fibrosis in mice to certain degree, suggesting that Chinese herbs with supplementing qi and activating blood circulation effect could play a definite preventive action on pulmonary fibrosis, and the inhibition on expressions of TGF-beta1 in lung tissues may be one of the mechanisms.

Total glucosides of Picrorhiza scrophulariiflora (TGP) is the active principal in Picrorhiza scrophulariiflora which has antioxidant effect. Since oxidative stress plays a key role in diabetic nephropathy, we investigated the effects of TGP on oxidative stress in bovine glomerular mesangial cells (MC) induced by prolonged high glucose. Bovine glomerular mesangial cells were cultured and passages 2-3 were used for the experiment. Mesangial cells were cultured in high glucose medium, and treated with TGP for 3 weeks. Then collagen IV excreted by mesangial cells were detected, and the percentages of cell cycle were observed by flow cytometry technique. The levels of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and [Ca2+]i were measured by flow cytometry after loaded with fluorescent probe DCFH-DA, Rh123 and fluo-3-acetoxymethylest. TGP significantly decreased the excretion of collagen IV and cell hypertrophy induced by high glucose, reduced the levels of ROS and [Ca2+]i, and increased MMP. Therefore we conclude that TGP could protect mesangial cells against oxidative stress induced by high glucose.

Objective: To observe the effect of tyrosine kinase depressor imatinib mesylate on pulmonary fibrosis (PF) induced by bleomycin in mice and explore its mechanism., Methods: One hundred and twenty C57BL/6 mice were randomly divided into control group, model group, dexamethasone group and imatinib mesylate group (with 30 mice each). After the model of PF was reproduced by a single intratracheal instillation of bleomycin, the above-mentioned drugs were given respectively. On 7, 14 and 21 days after treatment, 10 mice of each group were sacrificed, their lungs were harvested for measurement of the lung index, pathological change, concentration of hydroxyproline (HYP) and expression of platelet-derived growth factor-BB (PDGF-BB) in lung tissues., Results: Lung indexes of model group were significantly higher than those of control group at different time points after in tracheal bleomycin (all P<0.05). Alveolitis grades and PF degrees were elevated markedly (all P<0.01). The concentrations of HYP in lung tissues were continuously increasing with the elapse of time after bleomycin (all P<0.05). The contents of PDGF-BB were obviously higher than those of control group (all P<0.05), and they gradually reduced with time. In imatinib mesylate and dexamethasone groups alveolitis grades and PF induced by bleomycin were obviously alleviated. The concentration of HYP and expression of PDGF-BB in lung tissues decreased in imatinib mesylate and dexamethasone groups compared with those in model group (P<0.05 or P<0.01). The effects of imatinib mesylate on above indexes were better than those of dexamethasone but without significant differences (all P>0.05)., Conclusion: Imatinib mesylate may obviously inhibit the development of PF induced by bleomycin in mice.