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Antioxidant action is critical for maintaining the normal cardiovascular function and vascular endothelial cell is an important target of estrogen action through estrogen receptor (ER) pathway. This study is carried out to explore the antioxidant effect of carnosol in bovine aortic endothelial cells (BAECs) via ER pathway. The ER subtype specific estrogenic effect of carnosol was further demonstrated by luciferase reporter gene assay in human embryonic kidney (HEK) 293 cells. Carnosol was extracted from Chinese medicine Rosmarinus officinalis. ER positive BAECs were employed in cell proliferation assay and cell apoptosis tests. Oxidative stress by intracellular reactive oxygen species (ROS) were measured via 2'7'-dichlorofluorescein (DCF) production. ERα and ERβ specific antagonists 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidine-3-yl]phenol (PHTPP) were employed as tools in the experiment. ER negative HEK 293 cells were employed in luciferase reporter gene assay. The results indicate that carnosol can effectively attenuate H(2)O(2) induced slowing down of cell growth and increasing of cell apoptosis. At the meantime, carnosol pretreating can also effectively reduce the H(2)O(2) induced intracellular ROS elevation in BAECs. ERα and ERβ antagonist, especially ERα antagonist, can effectively decrease the above antioxidant effects of carnosol. The reporter gene analysis further demonstrates that the action of carnosol on inducing ERE dependent luciferase expression is realized via ER pathway. The conclusion is that carnosol can exert antioxidant effects towards oxidative stress induced by H(2)O(2) in BAECs. And such effects are realized via ER, especially ERα pathway. The results contribute to explain the mechanism of cardiovascular protective function of carnosol in postmenopausal women.

To study the protective effect of soybean protease inhibitor on LPS-induced lung injury in rats.Fifty male SD rats were randomly divided in five groups, 10 rats in each group as sham-operation group, model control group, positive medicine group, and high, moderate SBTI groups. Except the sham-group, other groups were induced by intratracheal instillation of LPS with a dose of 6 mg x kg(-1). All rats were given drug throughout intraperitoneal injection except the model controlled group, the positive medicine group was given PMSF with a dose of 50 mg x kg(-1), the high dose group of SBTI was given SBTI with a dose of 100 mg x kg(-1), a dose of the moderate group is 50 mg x kg(-1). We examined all rats in seven days. Index exam: cell quantity, activity of neutrophilic granulocyte released elastic protease proteins in BALF, histopathological examination and so on.Soybean protease inhibitor can level down the level of total protein, cell quantity, PMN percent, activity of neutrophilic granulocyte in BALF. SBTI level down the content of NF-kappa B in nucleoprotein, while increase the content of I kappa B alpha in plasmoprotein.SBTI is useful in protecting experimental pulmonary injury induced by LPS in rats.

To investigate the effects of bleomycin (BLM) on the apoptosis of type II alveolar epithelial cell (AT II) in lung fibrotic rats and its possible mechanisms.Totally 32 male Sprague-Dawley rats were randomly divided into sham group (n = 8) and BLM group (n = 24). Rats in sham group or BLM group were intratracheally instillated with saline or 5 mg/kg of bleomycin, respectively. One, three, and seven days after the instillation of bleomycin, 8 rats in BLM group were taken for AT II isolation and purification. Rats in sham group were used to isolate and purify AT II on 7 days after the instillation of saline. The cell cycle and apoptosis, intracellular free calcium concentration, and mitochondrial membrane potential (MMP) in AT II were determined by flow cytometry. Immunohistochemistry was performed to observe the expressions of Bax, Bcl-2, and Fas. Caspase-3, Caspase-8, and Caspase-9 activities were measured by Caspase activity detection kit.The ratio of S phase AT II in BLM group was significantly lower than in sham group (P0.05). AT II apoptosis rates on day 1 and 3 were significantly higher in BLM group than in sham group (P0.01). Intracellular free calcium concentrations in BLM group were significantly higher than in sham group (P0.05). However, MMP was significantly lower than sham group (P0.05). The positive rates of Bax, Fas and Caspase-3, Caspase-8, and Caspase-9 activities of BLM group were significantly higher than those of sham group (P0.05, P0.01). The positive rates of Bcl-2 on day 1 and 3 were significantly lower than those of sham group (P0.05).Early AT II apoptosis may be induced by bleomycin, which may be explained by the increase of intracellular free calcium concentration, depression of MMP, increased expressions of Fas and Bax, and increase of Caspase-3, Caspase-8, and Caspase-9 activities.

To explore the phytoestrogenic effects of ten kinds of Chinese medicine including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus ligustri lucidi, fructus lycii, radix clycyrrhizae, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae.240 female Kunming mice weighting 9 - 12 g were randomly divided into two main groups A and B. A group was divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine groups. B group was also divided into 12 small groups: 1 solvent control group, 1 diethylstilbestrol control group and 10 Chinese medicine antagonistic groups. Mice in ten antagonistic groups were administered both Chinese medicine and diethylstilbestrol everyday. After administered(op) for 4 days, blood was collected and serum was separated. The effect of the pharmacological serum on proliferation rate of MCF-7 (ER+) was analyzed by MTT-assay.In A group, proliferation rates of MCF-7 cells treated with serum from eight Chinese medicine groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, fructus lycii, herba cistanches, herba epimedii, fructus psoraleae and semen cuscutae were coued markedly increase respectively. While serum from fructus ligustri lucidi group could markedly decrease the proliferation rate of MCF-7 cells. In B group, the increased proliferation rate of MCF-7 cells caused by diethylstilbestrol was significantly reduced in seven Chinese medicine antagonistic groups including flos carthami, radix cyathulae, radix salviae miltiorrhizae, radix clycyrrhizae, herba epimedii, fructus psoraleae and semen cuscutae. While the increased proliferation rate could be markedly enhanced in herba cistanches group.Six kinds of Chinese medicine such as flos carthami, radix cyathulae, radix salviae miltiorrhizae, herba epimedii, fructus psoraleae and semen cuscutae show both estrogenic effects (when administered indepently) and antiestrogenic effects (when administered together with diethylstilbestrol). Such bidirectional effects depends on the internal estrogen level.

To observe the changes of matrix metalloproteinases (MMPs) activities in pulmonary fibrosis rats.Eighty male SD rats were randomly divided into sham group (n = 40) and bleomycin group (BLM, n = 40), in which SD rats were injected with a single intratracheal dose of sham saline or bleomycin respectively. On day 1, 3, 7, 14, and 28 following bleomycin or saline instillation, rats were randomly killed, and serum from abdominal aorta, alveolar fluid from the bronchoalveolar lavage, and the lung homogenate were collected and then stored at -80 degrees C. MMPs activity was determined by zymography.Compared with sham group, the levels of MMP-9 in all samples were augmented. MMP-9 activities in the serum were highest on day 3 than those on day 1 and day 7, and in lung tissue homogenate were highest on day 7; however, no significant differences were found between BLM group and sham group on day 14 and day 28; and that of bronchoalveolar lavage fluid (BALF) was highest on day 7 than those on day 1 and day 14, while no significant difference existed between BLM group and sham group on day 28. Serum MMP-2 level did not change from day 1 to day 28, while the level of BALF MMP-2 began to increase after day 14, even on day 28. Lung tissue homogenate MMP-2 level began to increase early on day 3 and continued to day 28.The sources and effects of MMP-2 and MMP-9 differ in BLM-induced rat pulmonary fibrosis.

To evaluate the estrogenic activity of several kinds of Chinese herbal medicines.Use zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.The botanical extracts of S. tamariscina can increase weights of mice (P0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.S. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.

To study the protective effects of curcumin on exaggerated extracellular matrix accumulation of pulmonary fibrosis rats.One hundred and forty-four male Sprague-Dawley rats were randomly divided into 6 groups (24 rats in each group). Rats in the model control group, positive medicine group, and high, moderate and low curcumin groups were injected with a single dose of bleomycin by trachea, and rats in sham-model control group with same volume normal saline. One day after the injection, curcumin solution of different dosages (200, 100, 50 mg x kg(-1) x d(-1)) was respectively given to rats in the high, moderate and low curcumin group daily by gastrogavage, while equal volume of normal saline was given to those in the sham-model control group and model control group, and an equal volume of prednisone (0.56 mg x kg(-1) x d(-1)) was given to those in positive medicine control group. On the 7, 14, 28 days, 8 rats per treatment group were randomly killed, the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum were determined, the determination of hydroxyproline in lung homogenates was analyzed, and the lung was incised to make pathological sections which were stained with HE and Mallory.Curcumin could decreas the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum, and inhihit the proliferation of fibrous tissue.Curcumin may play its therapetuic role by leveling down the content of extracellular matrix in rats with pulmonary fibrosis induced by bleomycin.

AIM: To study the role of hepatic sinusoidal capillarization and perisinusoidal fibrosis in rats with alcohol-induced portal hypertension and to discuss the pathological mechanisms of alcohol-induced hepatic portal hypertension. METHODS: Fifty SD rats were divided into control group (n=20) and model group (n=30). Alcoholic liver fibrosis rat model was induced by intragastric infusion of a mixture containing alcohol, corn oil and pyrazole (1 000:250:3). Fifteen rats in each group were killed at wk 16. The diameter and pressure of portal vein were measured. Plasma hyaluronic acid (HA), type IV collagen (CoIV) and laminin (LN) were determined by radioimmunoassay. Liver tissue was fixed in formalin (10%) and 6-μm thick sections were routinely stained with Mallory and Sirius Red. Liver tissue was treated with rabbit polyclonal antibody against LN and ColIV. Hepatic non-parenchymal cells were isolated, total protein was extracted and separated by SDS-PAGE. MMP-2 and TIMP-1 protein expression was estimated by Western blotting. RESULTS: The diameter (2.207 ± 0.096 vs 1.528 ± 0.054 mm, P

To explore the possible effects and mechanism of Fufang Biejiafang on a single intratracheal instillation (IT) of bleomycin-induced lung fibrosis model.SD rats were treated with a single IT dose of bleomycin or control saline. Chinese medicine group were poured into the stomach after the first day of operation with high dosage, middle dosage and low dosage. On days 7, 14 and 28 following IT bleomycin or saline, 4 mL blood were taken from the abdominal aorta for arterial blood gas analysis. The left lung was fixed for routine light microscopic examination. Bronchoalveolar lavage fluid (BALF) from the right lung was tested the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance, then the right lung was frozen immediately in liquid nitrogen for determination of hydroxyproline concentration.Model rats had obviously changes of body weight and hypoxemia and dysfunction of PS on days 7 and improved on days 14. Compared with three dose groups, the middle dose group some degreely improved and PS function. It ameliorate fibrosis because of inhibition of inflammation.(1) PS dysfunction resulted in hypoxemia after bleomycin injured alveolar type II (AT II). Fufang biejiafang-middle dose-group ameliorate hypoxemia by remission AT-II injury. (2) Fufang biejiafang may inhibit exudation inflammation and ameliorate fibrosis.

To study the influence of compound Biejia Ruangan Prescription (CBRP) on extracelluar matrix in bleomycin induced pulmonary fibrosis rats.54 male Sprague-Dawley rats were randomly divided into 6 groups (9 rats in each group). Rats in the model control group, positive medicine group, and high, moderate and low CBRP groups were injected with bleomycin A5 by trachea, and rats in sham-model control group with same volume normal saline. 29 days after the injection, CBRP solution of different dosages (1.4 g x kg(-1), 0.7 g x kg(-1), 0.35 g x kg(-1)) was respectively given to rats in the high, moderate and low CBRP group by gavage, while equal volume of normal saline was given to those in the sham-model control group and model control group, and an equal volume of prednisone (0.56 mg x kg(-1)) was given to those in positive medicine control group. On the 80th day, the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum were determined, the determination of hydroxyproline in lung homogenates was analyzed, and the right lung was incised to make pathological sections which were stained with Hematoxylin-Eosin (HE) and Masson staining for pathological diagnosis.CBRP could decrease the levels of III-collagen, IV-collagen, laminin and hyaluronic acid in the serum.CBRP may play its therapeutic role by leveling down the content of extracellular matrix in rats with pulmonary fibrosis induced by Bleomycin A5.

To study the effects of extracts of root of kudzu vine on mammary gland and uterus development in rats.40 Wistar rats weighting 65-85 g were randomly divided into 4 groups: control group, estrogen group, extracts of root of kudzu vine group of high dose, extracts of root of kudzu vine group of low dose. (10 rats in each group). After having been treated for 7 days, the rats were killed; mammary glands and uterus were removed and weighed. Serum was isolated and kept at 4 degrees C for determination of hormones.1. Administration of the root of kudzu vine significantly increased the weigh of mammary gland and uterus in rats. 2. Administration of the root of kudzu vine increased serum FSH, LH, E2 and decreased PRL.Extracts of root of kudzu vine could enhance the weight of mammary gland and uterus growth in rats, which may provide experimental evidence for the development of new drug used for promoting mammary gland and uterus.

In order to study on effects of soy extract on energy metabolims in ovariectomized rats.90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone of low dose, soy extract of high dose, soy extract of low dose, 10 rats each group. Beside of control and sham groups, the rest rats were ovariectomized. One week after operation, the rats were treatmented with different drugs, measument of body weigh and feed weigh each week. Six week after operation, the rats were killed, serum were taken, abdomen lipid were removed and weight.The ovariectomized rats took more food and got weight gain significantly; Body mess index(BMI), Abdomen lipid weigh and food transform rate in Model group increased significantly than control and sham groups. Administration of estrogen or soy extract or soy isoflavone could block these changes in ovariectomized rats, but soy polysaccharides did not have the effects.Ovariectomized rats have imbalance of energy metabolism, weigh gain and accumulation of abdomen lipid; administration of estrogen, soy extracts or soy isoflavone could attenuate these changes induced by ovariectomizing.

To study effects of soy extract on lipid metabolims in ovariectomized rats.90 Wistar rats were randomly divided into 9 groups: control group, sham group, model group, estrogen group, soy isoflavone group of high dose, soy isoflavone group of low dose, soy extract of high dose, soy extract of low dose, and soy polysaccharde group, 10 rats in each group. Except fer of control and sham groups, the test rats were ovariectomized. One week after operation, the rats were treated with different drugs. Six weeks after operation, the rats were killed, with serum and liver taken, and serumglycerol(sGT), cholesterol(sGC), LDL, HDL and liver homogenate hGT, hGC, measured.The level of sGC, LDL in ovariectmized rats increased significantly, compared with that in control and sham groups. In liver both the level of hGT and hGC were higher than that in liver from control and sham groups. Administration of estrogen or soy extract or soy isoflavone could attenuate these in ovariectomized rats, but soy polysacchardes did not have any effects.Ovariectomized rats have an imbalance of lipid metabolism, the level of hGT and hGC were increased, and administration of estrogen, soy extracts or soy isoflavone could decrease these changes induced by ovariectomizing.

AIM: To study the effects of Radix Puerariae flavones (RPF) on liver lipid metabolism in ovariectomized (OVX) rats. METHODS: Forty adult female Wistar rats were randomly divided into four groups: OVX group; sham-OVX group; OVX + estrogen group and OVX + RPF group. One week after operation rats of the first two groups were treated with physiological saline, rats of OVX + estrogen group with estrogen (1 mg/kg·b.w.) and rats of OVX + RPF group with RPF (100 mg/kg·b.w.), respectively for 5 weeks. After the rats were killed, their body weight, the weight of the abdominal fat and uterus were measured, and the levels of total cholesterol (TC) and triglyceride (TG) in liver homogenate were determined. RESULTS: Compared with the sham-OVX group, the body mass of the rats in OVX group was found increased significantly; more abdominal fat in store; TC and TG in liver increased and uterine became further atrophy. As a result, the RPF was found to have an inhibitive action on those changes of various degrees. CONCLUSION: RPF has estrogen-like effect on lipid metabolism in liver and adipose tissue.

AIM: To observe the effect of compound Biejiaruangan decoction (CBJRGC) (composite prescription of Carapax trionycis for softening the liver) on proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells (HSCs) and to find the mechanism of prevention and treatment of hepatic fibrosis by CBJRGC. METHODS: Using MTT, immunohistochemistry and image analysis technology, the related indexes for proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells were detected in 24 h, 48 h, and 72 h after adminstration of different dosages of CBJRGC. RESULTS: Statistical analysis showed that serum collected from rat perfused with CBJRGC could restrain the proliferation of HSC in 48 h and 72 h especially in high and medium dosage groups, markedly decrease the expression of desmin, synapsin and platelet derived growth factor (PDGF) in HSC in 24 h, 48 h and 72 h, as well as the expression of α-SMA, collagen III, TIMP and TGFβ1 in 48 h and 72 h, decrease the excretion of collagen I in 72 h. CBJRGC serum had no significant effect on collagens I, III and TIMP in 24 h. CONCLUSION: CBJRGC serum has a good curative effect on hepatic fibrosis. Its main mechanism may be related to the following factors. The drug serum can restrain the proliferation and activation of HSC, decrease the number of activated HSC and the total number of HSC, the excretion of collagens I, III, enhance the degradation of collagen and restore the balance of synthesis and degradation of collagen, inhibit the expression of transforming growth factor β1 (TGFβ1) and platelet derived growth factor (PDGF) in HSC, block and delay the process of hepatic fibrosis. Synapsin is a new marker of activation of HSC, which provides a theoretical and testing basis for neural regulation in the developing process of hepatic fibrosis.