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This study aimed to investigate the structure of a new heteropolysaccharide (MC-Pa) from Moutan Cortex (MC), and its protection on diabetic nephropathy (DN). The MC-Pa composed of D-glucose and L-arabinose (3.31:2.25) was characterized with homogeneous molecular weight of 1.64 × 105 Da, and the backbone was 4)-α-D-Glcp-(1 → 5-α-L-Araf-(1 → 3,5-α-L-Araf-(1→, branched partially at O-3 with α-L-Araf-(1 → residue with methylated-GC–MS and NMR. Furthermore, MC-Pa possessed strong antioxidant activity in vitro and inhibited the production of ROS caused by AGEs. In vivo, MC-Pa could alleviate mesangial expansion and tubulointerstitial fibrosis of DN rats in histopathology and MC-Pa could decrease significantly the serum levels of AGEs and RAGE. Western blot and immunohistochemical analysis showed that MC-Pa can reduce the expression of main protein (FN and Col IV) of extracellular-matrix, down-regulate the production of inflammatory factors (ICAM-1 and VCAM-1), and therefore regulate the pathway of TGF-β1. The above indicated that MC-Pa has an improving effect on DN.

Copper nanoparticles assembled on carboxymethylcellulose (Cu0NPs@CMC) were successfully synthesized and well characterized by FT-IR, SEM, EDS, TEM, XPS, and ICP-AES. The new prepared nanocatalyst was applied effectively as a heterogeneous catalyst for the synthesis of propargylamines via decarboxylatived A3 and classic A3 reaction under solvent-free condition. A broad spectrum of diversely polysubstituted propargylamines could be obtained in moderate to excellent yields. The present method showed several merits such as easy work-up, short reaction time, additive-free characteristic, solvent-free condition, functional group tolerance, usage of recyclable green and sustainable nanocatalyst.

An ordinary silica gel (SG-1) was chosen as the raw material and hydrophobically modified with trimethylchlorosilane under different microwave exposure. The orthogonal experiment was designed to screen the optimized modification method. The hydrophobic properties modified silica gel (SG-2) was analyzed by comparing the difference changes of surface chemistries. Then, the adsorption capacities of SG-1 and SG-2 to the n-hexane vapor and water vapor, respectively, were investigated and compared with the capacities of activated carbon and SG-3 (hydrophobically modified using the same composite modifier without microwave exposure). Meanwhile, there cyclable adsorptions of SG-2 to the two vapors were conducted while the desorptions of vacuum and heat was observed. The results showed that the effect of hydrophobic modification to SG-1 under microwave exposure was obvious and SG-2 also had a very good effect of adsorption for the n-hexane vapor. The modified silica gel shows great potential and effect for the recovery of volatile organic compounds with high content of water.

Ethnopharmacological relevance Epimedium koreanum Nakai is documented as tonic herbal in China for over a thousand years and has the potential to enhance the body's immunity according to the theory of traditional Chinese medicine. Polysaccharides are one of the most important effective compounds in Epimedium koreanum Nakai. Accumulating evidence indicated polysaccharides derived from traditional Chinese medicine have potent immune-enhancing properties and relatively nontoxic effects in cancer treatment. However, information about immunological regulation in tumor of Epimedium koreanum Nakai polysaccharides is limited and the reports of purification, characterization of polysaccharides have remained less. The purpose of our study was to further investigate the active polysaccharides from Epimedium koreanum Nakai by evaluating the immune-regulation activities in tumor-bearing mice and provide reasonable explanation for traditional application. Materials and methods We firstly purified Epimedium koreanum polysaccharide (EPS) from crude extracts and evaluated EPS in vitro using immunological experiments including maturation and Ag presentation function of DCs, CD4 T-cell differentiation and secretion of anti-cancer cytokines. In LLC-bearing mice model, we investigated its antitumor activities through evaluation of tumor cell proliferative activity, calculation of immune organ indexes and relative host immune system function tests. Results Results showed that EPS (180 × 104 Da) was composed of mannose (Man), rhamnose (Rha), glucuronic acid (GlcUA), galactosamine (GalN), glucose (Glc), galactose (Gal), arabinose (Ara) and fructose (Fuc). Chemical composition assay indicated EPS was a fraction with 28.20% uronic acid content. FT-IR suggested the presence of pyraoid ring in EPS and SEM displayed smooth surface embedded by several pores. Moreover, Our study suggested EPS could remarkably stimulate macrophages to secrete substantial anti-cancer cytokines and promote maturation as well as Ag presentation function of DCs. Strikingly, CD4 T-cell differentiation and increased INF-γ production stimulated by EPS–activated macrophages were observed in the research. Furthermore, EPS exhibited prominent antitumor activities through regulating host immune system function in LLC-bearing mice. Taken together, experimental findings suggested EPS could be regarded as a potential immune-stimulating modifier for cancer therapy. Conclusion Our studies demonstrated the polysaccharide (180 × 104 Da) purified from Epimedium koreanum Nakai could promote maturation and Ag presentation function of DCs, increase the level of immunomodulatory cytokines and activate CD4 T-cell differentiation. Furthermore, it may inhibit the tumor growth in LLC-bearing mice through regulating host immune system function

The aim of this study was to establish a paclitaxel (PTX)-loaded mixed micelle delivery system (PTX-TP-M) with vitamin E-TPGS (TPGS) and Plasdone®S-630 Copovidone (PVPS630) as carriers to improve the solubility, oral absorption, and anti-tumor activity of PTX against lung cancer. In this study, PTX-TP-M was prepared using the ethanol thin-film dispersion method followed by characterization of the binary mixed micelles system. The average size of the PTX-TP-M was 83.5 ± 1.8 nm with a polydispersity index of 0.265 ± 0.007 and the drug loading (DL%) and entrapment efficiency (EE%) were 3.09 ± 0.09% and 95.67 ± 2.84%, respectively, which contributed to a high solubility of PTX about 24947-fold increase in water (4.78 ± 0.14 mg/mL). In addition, TEM analysis showed that the PTX-TP-M appeared spherical in structure and was well dispersed without aggregation and adhesion. In vitro release studies showed that the PTX-TP-M displayed a sustained release compared to free PTX in the dialysis bag. The efflux ratio of PTX reduced from 44.83 to 3.52 when formulated as PTX-TP-M; a 92.15% reduction, studied using the Caco-2 monolayer model. The oral bioavailability of PTX also improved by 4.35-fold, suggesting that PTX-TP-M can markedly promote the absorption in the gastrointestinal tract. Using in vitro MTT assays, it was observed that cytotoxicity was markedly increased, and IC50 values of PTX-TP-M (3.14 ± 0.85 and 8.28 ± 1.02 μg/mL) were lower than those of PTX solution (5.21 ± 0.93 and 14.53 ± 1.96 μg/mL) in A549 and Lewis cell, respectively. In vivo anti-tumor studies showed that PTX-TP-M achieved higher anti-tumor efficacy compared with PTX in Lewis bared C57BL/6 mice. Furthermore, a gastrointestinal safety assay also proved the safety of PTX-TP-M. All results demonstrated that the PTX-TP-M exhibited great potential for delivering PTX with increased solubility, oral bioavailability, and anti-cancer activity and this binary mixed micelles drug delivery system has potential to be used clinically.

The aim of this study was to investigate the effect of a polysaccharide from Scutellaria baicalensis Georgi on UC. Gut microbiota dysbiosis is a worldwide problem associating with ulcerative colitis. One homogeneous polysaccharide, named SP2-1, was isolated from Scutellaria baicalensis Georgi. SP2-1 comprised mannose, ribose, rhamnose, glucuronic acid, glucose, xylose, arabinose, fucose in the molar ratio of 5.06:21.24:1.00:20.25:3.49:50.90:228.77:2.40, with Mw of 3.72 × 106 Da. SP2-1 treatment attenuated body weight loss, reduced DAI, ameliorated colonic pathological damage, and decreased MPO activity of UC mice induced by DSS. SP2-1 also suppressed the levels of proinflammatory cytokines. Additionally, the intestinal barrier was repaired due to the up-regulated expressions of ZO-1, Occludin and Claudin-5. SP2-1 remarkably enhanced the levels of acetic acid, propionic acid, and butyric acid in DSS-treated mice. Furthermore, as compared with model group, the abundance of Firmicutes, Bifidobacterium, Lactobacillus, and Roseburia were significantly increased with SP2-1 treatment. And SP2-1 could significantly inhibit the levels of Bacteroides, Proteobacteria and Staphylococcus. In conclusion, SP2-1 might serve as a novel drug candidate against UC.

The purpose of the study was to extract, separate, purify and character Scutellaria baicalensis Georgi polysaccharides, and to further explore the potential mechanism on colitis. Scutellaria baicalensis Georgi was extracted with water and precipitated with alcohol. The obtained crude polysaccharide was fractionated sequentially by DEAE-52 ion exchange column and Sephadex G-100 gel colum. Then, we obtain the purified fraction, named SP1-1. SP1-1 (4.56 × 105 Da) was mainly composed of mannose, ribose, glucuronic acid, glucose, xylose and arabinose with molar ratios of 2.14:3.61:1:2.86:5.98:36.39. FI-TR indicated SP1-1 had the characteristic absorption of polysaccharides with a pyranoid ring. SEM displayed honeycomb structure. Oral administration of SP1-1 significantly decreased disease activity index, ameliorated dextran sulfate sodium-induced colonic pathological damage, and reduced myeloperoxidase activity. Additionally, SP1-1 markedly suppressed the proinflammatory cytokines level, including IL-1β, IL-18 and TNF-α, in the serum and colon of DSS-induced mice and THP-1-derived macrophages. Furthermore, a decreased CD11b+ macrophage infiltration in colons and inactivation of caspase-1 in peritoneal macrophages were detected in SP1-1-treated mice. The mechanisms responsible for these effects of SP1-1 were attributed to its inhibition on NF-κB signaling and NLRP3 inflammasome activation. These findings support SP1-1 as a novel drug candidate in the treatment of colitis.

Background. Phyllanthus emblica L (PEL), a well-known medical plant, has been used in Asian countries for a long time. Increasing evidence suggests that it can prevent the tumorigenesis of cancer associated with nonresolving inflammation. However, the possible anti-inflammatory mechanism responsible for preventing tumorigenesis of precancerous lung lesions is not well elucidated. Materials and methods. Male A/J mice were randomly divided into 5 groups with 10 mice in each group: (1) blank group (saline), (2) benzo(a)pyrene [B(a)P] group, (3) and (4) B(a)P + PEL (5 g/kg/d, 10 g/kg/d, administered by gavage), (5) B(a)P + celecoxib (30 mg/kg/d, administered by gavage). Nodes on the lung surface were observed and calculated. The levels of macrophage inflammatory protein (MIP-2), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA) kits. Cyclo-oxygenase-2 (COX-2), hypoxia-inducible factor-1 (HIF-α), IL-1β, miR-101, and Lin28B protein levels were evaluated by immunohistochemistry and Western blotting. Results. PEL extract treatment significantly reduced the number of nodes on the lung surface and attenuated B(a)P-induced levels of proinflammatory cytokines MIP-2, TNF-α, IL-6, and IL-1β in lung tissue. The protein expressions of COX-2 and HIF-α were significantly decreased by the treatment of PEL. In addition, both PEL extract and celecoxib markedly upregulate the expression of miR-101 while downregulating IL-1β and Lin28B levels. Conclusion. Our study indicated that treatment with PEL extract can not only protect the lung from inflammatory injury but effectively prevent precancerous lung lesions through regulating the IL-1β/miR-i101/Lin28B signaling pathway.

Ethnopharmacological relevance Combination of Ligusticum chuanxiong and Radix Paeoniae (XS) is highly effective in the treatment for focal cerebral ischemic, but the underlying mechanism is not clear. This study was conducted to evaluate the combinative effects of XS on MCAO rats and explore the underlying mechanisms. Materials and methods MCAO rats were used to evaluate the protective effect of Ligusticum chuanxiong (CX), Radix Paeoniae Rubra (CS) and their combination (XS) on ameliorating focal cerebral ischemic. Cerebral ischemia deficits and infarct size were performed by 2,3,5-triphenyltetrazolium chloride (TTC) and hematoxylin-eosin (H-E) staining. Activities of SOD, CAT and GSH-Px, as well as levels of LPO and MDA were detected by commercial kits while ELISA kits for the content of plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator (PA). Immunohistochemistry (IHC) and western blot analysis (WB) were carried out to examine the protein expressions including PKR-like endoplasmic reticulum kinase (PERK), cytoplasmic of glucose regulated protein 78 (GRP78), X box-binding protein-1 (XBP-1), activating transcription factor-6 (ATF-6), C/EBP-homologous protein (CHOP), metalloprotease-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), Bcl-2 associated X protein (Bax), and porcineB-cellleukemia/lymphoma-2 (Bcl-2) in brain tissues. Reverse transcription polymerase chain reaction (RT-PCR) and Quantitative PCR (Q-PCR) were applied to examine vascular endothelial growth factor (VEGF) and N -methyl- d -aspartate receptors (NMDAR1) mRNA levels. Results CX, CS and their combination (XS) could reduce cerebral ischemia deficits and infarct size of MCAO rats. They increased SOD, CAT and GSH-Px activities, and reduced MDA and LPO levels in serum, markedly. A significant decrease of endoplasmic reticulum stress-related factors PERK, XBP-1, ATF-6 and CHOP protein expression levels while an increase of GRP78 and MVD expression by the treatment of CX, CS and XS. It could also be observed that their treatment could reduce apoptotic damage of brain tissues by up-regulating Bax level and down-regulating Bcl-2 level. Furthermore, the levels of MMP-9 and PAI-1 in serum and tissues of rats were down-regulated remarkably while TIMP-1 and PA levels were up-regulated. VEGF mRNA level was up-regulated dramatically whereas NMDAR1 was reduced. Importantly, the combination of CX and CS, namely XS, has a more meaningful improvement on focal cerebral ischemic than CX or CS alone. Conclusion All these revealed that the combined XS exerted more remarkable protective effects than alone. XS could inhibit neuronal apoptosis by attenuating ER-stress-dependent apoptotic signaling and protected the blood-brain barrier. These findings might supply beneficial hints for the synergy of CX and CS, and provide the basis for rationality of XS preparation and deserve further clinical investigations.

Excessive autophagy plays a crucial role in cigarette smoking extract (CSE)-induced inflammation response and oxidative damage of respiratory epithelial cells. The components from Eclipta prostrata L. (CCE) have been shown to be beneficial for CSE-induced epithelial cells injury. However, whether its protection on CSE-stress injury is related to its regulation on autophagy remains still unclear. In this study, CCE, containing mainly wedelolactone of 45.88% and demethylwedelolactone of 23.74%, could improve significantly 10%CSE-induced cell viability of normal human bronchial epithelial (NHBE) cells using CCK-8 kit. We revealed that CCE could remarkably increase autophagic factors Beclin-1, Atg5, ATF4 proteins expression levels and the transformation of LC3-I to LC3-II. Additionally, CCE up-regulated significantly p-p16 and p-p21 phosphorylation levels whereas down-regulated p-p53 in NHBE cells. The changes of typical autolysosom and representative autophagosome in the presence of CCE or/and autophagy inhibitor chloroquine (CQ) were also observed by transmission electron microscopy. These data demonstrated that CCE reduced CSE-induced autophagy flux activation in NHBE cells. The blockade of CCE on autophagy flux contributes to its protection against CSE-induced NHBE cells damage, and CCE is promising to be combination therapeutic molecules to excessive autophagic damage in respiratory diseases.

Fraxini Cortex (also known as Qinpi, QP) has been used for the treatment of hyperuricemia with a significant difference on efficacy of QP from different regions. However, it`s still unknown whether proportion of components is the key and why same kind of herbs have different therapeutic effects. In this study, different sources of QP were collected from Shaanxi Qinpi extracts (SQPE), Henan Qinpi extracts (HQPE), Hebei Qinpi extracts (GQPE) provinces in China. Rat model of hyperuricemia with hypoxanthine combined with potassium oxonate were established to determine the levels of blood urea nitrogen (BUN), serum uric acid (SUA), urine uric acid (UUA) and creatinine (Cr). Hematoxylin-eosin staining (HE) and Periodic Acid-Schiff staining (PAS) were performed for renal pathology while Western blot analysis and real-time PCR analysis for proteins and mRNA expression levels. High-performance liquid chromatograph (HPLC) was used for components and composition analysis. Our results demonstrated that QPE from different regions could alleviate hyperuricemia via increasing significantly the SCr and BUN levels whereas decreasing markedly UCr, SUA and UUA levels. Additionally, QPE could also improve the pathological changes of the kidneys. The protein and mRNA levels of urate reabsorption transporter 1 (URAT1) and glucose transporter 9 (GLUT9) were down-regulated by QPE treatment. SQPE hold a better activity on improving hyperuricemia and regulating URAT1 and GLUT9. HPLC analysis showed that the proportion of four components aesculin, aesculetin, fraxin, fraxetin were 9.002: 0.350: 8.980: 0.154 (SQPE); 0.526: 0.164: 7.938: 0.102 (HQPE); 12.022: 1.65: 0.878: 1.064 (GQPE). These data indicate that this proportion of effective components may be an important factor for efficacy of QP and had implications for the treatment of hyperuricemia.

Goutengsan, a Chinese herbal formula, potential protection on Alzheimer’s disease (AD) has been less reported. In current study, we investigated the protection of Goutengsan on Aβ1–42-induced pheochromocytoma-derived cells (PC12). Furthermore, the components from Goutengsan in rat plasma were identified by microdialysis (MD) forin vivosampling. Meanwhile, the protection of components identified was also verified. At last, we found that Goutengsan has a potential protective effect on Aβ1–42-induced PC12 cells via reducing cells damage and increasing cells vitality as well as six components (pachymic acid, liquiritin, rhynchophylline, isorhynchophylline, corynoxeine, and isocorynoxeine) which may be effective components. This study helps to understand the treatment of Goutengsan for AD and would facilitate the clinical and further studies for this formula.

Epimedin C is one of the major bioactive constituents of Herba Epimedii. In this study, the metabolite profiles of epimedin C in rat plasma and bile were qualitatively investigated, and the possible metabolic pathways of epimedin C were subsequently proposed. After oral administration of epimedin C at a single dose of 80 mg/kg, rat biological samples were collected and pretreated by protein precipitation. Then these pretreated samples were injected into an Acquity UPLC BEH C18 column and detected by ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry. In all, 12 metabolites were identified in the biosamples. Of these, eight, two from plasma and six from bile, are, to our knowledge, reported here for the first time. The results indicated that epimedin C was metabolized via desugarization, dehydrogenation, hydrogenation, dehydroxylation, hydroxylation, demethylation and glucuronidation pathways in vivo. Thus, this study revealed the possible metabolite profiles of epimedin C in rat plasma and bile.

UNLABELLED Luteolin, one of the most common abundant flavonoids in vegetables and herbs, has antitumor effects on various tumors by inducing apoptosis, antioxidant effects and inhibition of angiogenesis. However, the potential chemoprevention of luteolin on lung cell damage and its related mechanism(s) are not fully known. The present study evaluated the protective effects of luteolin on cigarette smoke extract (CSE)-induced toxicity and apoptosis in normal human bronchial epithelial (NHBE) cells and explored its underlying mechanism(s). MTT assay showed that pretreatment with luteolin increased CSE-decreased cell viability (p

Graphical abstract, Phenolic acids, the main active ingredients in Flos Lonicerae extract possess strong antibacterial, antioxidant and antiviral effects, and their contents was higher largely than that of other ingredients such as flavones, but the absolute bioavailability orally was significantly low, which is significant low influencing clinical efficacies of its oral preparations. In the present study, in vitro Caco-2 cell, in situ single-pass intestinal perfusion and in vivo pharmacokinetics study were performed to investigate the effects of COS on the intestinal absorption of phenolic acids. The pharmacological effects such as antiviral activity improvement by COS were verified by MDCK cell damage inhibition rate after influenza virus propagation. The observations from in vitro Caco-2 cell showed that the absorption of phenolic acids in Flos Lonicerae extract could be improved by COS. Meanwhile, COS at the same low, medium and high concentrations caused a significant, concentration-dependent increase in the Papp-value for phenolic acids compared to the control group (p

Advanced glycation end products (AGEs) were implicated in the pathogenesis of endothelial dysfunction in diabetic vascular complications. Our previous study found that a novel compound 4,4'-diphenylmethane-bis(methyl) carbamate (CM1) from Cortex Mori (Morus alba L.) could attenuate AGE-induced endothelial dysfunction. The present study was conducted to explore the possible protective mechanisms of CM1 on AGE-induced endothelium damage. In binding experiments, fluorescence quenching and fluorescence polarization assays showed no significant difference or changes of AGEs on fluorescence intensity and polarization in the absence/presence of CM1. In AGE formation experiments, CM1 was incubated with AGE precursor compounds methylglyoxal (MGO), glyceraldehydes (Glycer) and glycolaldehyde (Glycol) in the formation system. However, high performance liquid chromatography (HPLC) analysis showed no new conjugated compounds formed in the reaction system. The results of ELISA analysis also showed that CM1 did not inhibit the AGE formation. However, the pretreatment with CM1 could significantly decrease AGE or high-mobility group box-1 (HMGB1, a ligand of RAGE)-induced cytotoxicity, apoptosis and reactive oxygen species (ROS) in human umbilical vein endothelial cells (HUVECs). Our results suggested that CM1 might block the AGEs-RAGE signal transduction rather than inhibit AGE formation or bind to AGEs and change its structure to prevent endothelial dysfunction in diabetic vascular complications.

In this work, the metabolite profiles of epimedin B in rat feces, bile, urine, and plasma were qualitatively investigated, and the possible metabolic pathways of epimedin B were subsequently proposed. After oral administration of epimedin B at a single dose of 80 mg/kg, rat biological samples were collected and pretreated by protein precipitation. Then, these pretreated samples were injected into an Acquity ultraperformance liquid chromatography BEH C₁₈ column with mobile phase consisting of 0.1% formic acid-water and 0.1% formic acid-acetonitrile and detected by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry. In all, 43 metabolites were identified in the biosamples. Of these, 13, including F5, F7, F16-F18, D5-D7, D9, N5, N7, M1, and M3, were to our knowledge reported for the first time. The results indicated that epimedin B was metabolized via desugarization, dehydrogenation, hydrogenation, hydroxylation, demethylation, glucuronidation, and glycosylation pathways in vivo. Specific hydrolysis of 7-O-glucosides in the gut lumen and glucuronic acid conjugation in the liver were considered as the main physiologic processes of epimedin B. This study revealed the possible metabolite profiles of epimedin B in rats.

The purpose of this work was to research the enhancement of Epimedium fried with suet oil based on the in vivo formation self-assembled flavonoid nanomicelles. Taking icariin as the representative, under the action of suet oil, self-assembled nanomicelles were prepared under simulated gastrointestinal tract conditions and were characterized by dynamic light scattering and transmission electron microscopy (TEM). The experiments with icariin self-assembled nanomicelles without suet oil were done according to the above. The influence of suet oil on the transportation of icariin across Caco-2 cell monolayers and the absorption in rat intestine of self-assembled nanomicelles were evaluated. The particle size of icariin self-assembled nanomicelles with suet oil was smaller than without suet oil. The nanomicelles seemed to be monodisperse spherical particle with smooth surfaces. The icariin entrapment efficiency of self-assembled nanomicelles with suet oil was increased from 43.1% to 89.7%. In Caco-2 cell monolayers, the absorptive permeability, secretory permeability and efflux ratio of icariin self-assembled nanomicelles with suet oil was 1.26#215; 10#8722;6 cm/s, 5.91#215; 10#8722;6 cm/s and 4.69, respectively, while that of icariin self-assembled nanomicelles without suet oil was 0.62#215; 10#8722;6 cm/s, 3.00#215; 10#8722;6 cm/s, and 4.84, respectively. In rat intestinal perfusion experiments, the permeability coefficient of icariin self-assembled nanomicelles with suet oil in duodenum was higher than the value of icariin self-assembled nanomicelles without suet oil (plt; 0.05). With the action of suet oil, icariin self-assembled nanomicelles were more stable and the entrapment efficiency was higher than that without suet oil, which could increase the solubility of icariin and improve its intestinal absorption. Therefore, suet oil plays a role in its enhancement.

In this work, the metabolite profiles of icariin in rat feces, bile and urine were qualitatively investigated, and the possible metabolic pathways of icariin were subsequently proposed. After oral administration of icariin at a single dose of 100 mg/kg, rat biological samples were collected and pretreated. Then, these pretreated samples were detected by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). In all, 17 metabolites were identified in the biosamples. Of these, 5, including F8-F9 (icariside I), D3-D4 (isopentenyl alcohol-icaritin-3-O-rha-7-O-gluA) and N3 (1,3-isoprene icariside II), were to our knowledge reported for the first time. The results indicated that icariin was metabolized via desugarization, dehydrogenation, hydroxylation, demethylation and glucuronidation pathways in vivo. This study revealed the possible metabolite profiles of icariin in rats.

The Chaihu-Shaoyao drug pair (Bupleuri Radix and Paeoniae Radix Alba) which is a traditional Chinese drug pair, has been widely used for anti-inflammatory purposes. Saikosaponin a (SSA), saikosaponin d (SSD) and paeoniflorin are identified as the main components in the pair. The present study focused on the interaction of the main components based on investigating their intestinal absorption using a four-site perfused rat intestinal model in order to clarify the mechanism of the compatibility of Chaihu-Shaoyao. The concentrations of SSA, SSD and paeoniflorin in the intestinal perfusate were determined by LC/MS or UPLC (Ultra Performance Liquid Chromatography) methods, followed by P*(eff) (effective permeability) and 10% ABS (the percent absorption of 10 cm of intestine) calculations. The results showed that all of the three main components displayed very low permeabilities (P*(eff) < 0.4), which implied their poor absorption in the rat intestine. The absorption levels of SSA and SSD were similar in intestine and higher in ileum than those in other intestinal regions in the decreasing order: colon, jejunum and duodenum. However, there is no significant difference in the absorption of paeoniflorin in the four segments (P < 0.05). The P*(eff) values of paeoniflorin exhibited an almost 2.11-fold or 1.90-fold increase in ileum when it was co-administrated with SSA and SSD, as well as 2.42-, 2.18-fold increase in colon, respectively, whereas the absorptions of SSA and SSD were not influenced by paeoniflorin. In conclusion, SSA and SSD could promote the absorption of paeoniflorin. To some extent this might explain the nature of the compatibility mechanisms of composite formulae in TCMs.

Cigarette smoke is the leading cause of the development of various lung diseases including lung cancer through triggering oxidant stress and inflammatory responses which contributed to the lesions of normal human bronchial epithelial (NHBE) cell. Wedelolactone (WEL), a natural compound from Eclipta prostrata L., has been found to possess the inhibitive effects on the proliferation and growth of cancers. In the present study, we investigated the effects of WEL on NHBE cell injury induced by cigarette smoke extract (CSE) in vitro. It showed that the pretreatment WEL (2.5-20μM) resulted in a significant protective effect on 10% CSE-induced cell death in NHBE cells. The pretreatment with WEL dose-dependently and significantly reversed the activities of SOD, CAT, GSH and the level of MDA to normal level. We also found that the protein expression levels of COX-2 and ICAM-1 which are related to inflammatory response were remarkably reduced by WEL compared with 10% CSE treatment. Additionally, WEL also reduced the expressions of antioxidases including NAD(P)H dehydrogenase:Quinone 1 (NQO1) and heme oxygenase-1 (HO-1). Moreover, Nrf2 inhibitor all-trans-retinoic acid (ATRA) decreased remarkably their expressions. These results suggest that WEL protects NHBE cell against CSE-induced injury through modulating Nrf2 pathway. Our study indicates that WEL may be a new potential protective agent against CSE-induced lung injury.

The Flos Lonicerae-Fructus Forsythiae herb couple is the basic components of Chinese herbal preparations (Shuang-Huang-Lian tablet, Yin-Qiao-Jie-Du tablet and Fufang Qin-Lan oral liquid), and its pharmacological effects were significantly higher than that in Flos Lonicerae or Fructus Forsythiae, but the reasons remained unknown. In the present study, pattern recognition analysis (hierarchical cluster analysis (HCA) and principal component analysis (PCA)) combined with UHPLC-ESI/LTQ-Orbitrap MS system were performed to study the chemical constitution difference between co-decoction and mixed decoction in the term of chemistry. Besides, the pharmacokinetics in vivo and intestinal absorption in vitro combined with pattern recognition analysis were used to reveal the discrepancy between herb couple and single herbs in the view of biology. The observation from the chemical view in vitro showed that there was significant difference in quantity between co-decoction and mixed decoction by HCA, and the exposure level of isoforsythoside and 3, 5-dicaffeoylquinic acid in co-decoction, higher than that in mixed decoction, directly resulted in the discrepancy between co-decoction and mixed decoction using both PCA and HCA. The observation from the pharmacokinetics displayed that the exposure level in vivo of neochlorogenic acid, 3, 4-dicaffeoylquinic acid, isoforsythoside and forsythoside A, higher than that in single herbs, was the main factor contributing to the difference by both PCA and HCA, interestingly consistent with the results obtained from Caco-2 cells in vitro, which indicated that it was because of intestinal absorption improvement of neochlorogenic acid, 3, 4-dicaffeoylquinic acid, isoforsythoside and forsythoside A that resulted in a better efficacy of herb couple than that of single herbs from the perspective of biology. The results above illustrated that caffeic acid derivatives in Flos Lonicerae-Fructus Forsythiae herb couple could be considered as chemical markers for quality control of its preparations.

The possible metabolic pathways of icariside II were proposed. An ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry method was used for analysing the faecal, bile, plasma and urine samples of rats administrated with icariside II. In all, 27 metabolites were identified in the biosamples. Of these, 20, including F1–F12, D3, D4, D6, D7–D9 and M3, M4, were, to our knowledge, reported for the first time. The results indicated that icariside II was metabolised via desugarisation, dehydrogenation, hydrogenation, hydroxylation, demethylation, glucuronidation, dehydration and glycosylation pathways in vivo. Specific hydrolysis of 7-O glucosides in the gut lumen and glucuronic acid conjugation in the liver were considered as the main physiologic processes of icariside II. This study revealed the possible metabolite profiles of icariside II in rats.

OBJECTIVE To investigate the protective effect and mechanism of Ecliptae Herba extract on cigarette smoke extract-induced cytotoxicity. METHOD The effect of Ecliptae Herba extract on CSE-induced NHBE cell proliferation was detected by MTT assay. GSH content was determined by DTNB colorimetry. GST activity was measured by CDNB colorimetric assay. NQO1 activity was detected by NADPH and DCIP. The protein expression was determined by Western blot assay. RESULT Ecliptae Herba extract reduced CSE's inhibitory effect on NHBE cells, recover the decrease in intracellular GSH caused by CSE and reduce the CSE-induced activity of GST and NQO1 and NQO1 protein expression. CONCLUSION Ecliptae Herba extract can reduce CSE-induced injury on NHBE cells, which may be related to phase II detoxification enzymes.

To study the effects of triterpenoid components from Prunella asiatica on phase II detoxifying enzymes and protein expression in vitro and in vivo. Normal human bronchial epithelial (NHBE) cell model was used in vitro, and the mouse model of Kunming (KM) mice was used in vivo. CDNB assay was used to measure the activity of GST. NADPH and DCIP was used to detect the activity of NQO1. DTNB colorimetric assay was used to detect GSH. Western blot was use to detect the protein expression of NQO1. We found that triterpenoid components from P. asiatica could increase the activity of GST, NQO1 and GSH in NHBE cells and KM mice. NQO1 protein expression can also be increased in vitro. The study suggests that triterpenoid components from P. asiatica can prevent the lung cancer by regulating the body phase II detoxification enzyme activity and protein expression.

Ethnopharmacological relevance Licorice (Glycyrrhiza uralensis roots) is used as a traditional medicine for the treatment of diabetes mellitus and its vascular complications. Glycyrrhizic acid (GA, also known as Glycyrrhizin), a triterpenoid saponin glycoside, is considered to be a bioactive component in Licorice and is beneficial to diabetic vascular complications. Aim of study The present study was conducted to evaluate the potential protective activities on AGEs-induced endothelial dysfunction, including anti-apoptosis, antioxidant stress and anti-proinflammatory responses, and explore the underlying mechanism. Materials and methods Human umbilical vein endothelial cells (HUVECs) were incubated and pre-treated with GA (10−9–10−6 M) or RAGE-Ab (5 μg/ml) in the presence or absence of 200 μg/ml AGEs. AO/EB fluorescence staining assay was performed to evaluate anti-apoptosis activity. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in cell supernatant were detected by kits while the intracellular reactive oxygen species (ROS) generation was determined by 2,7-dichlorodihydrofluorescin diacetate (DCFH-DA) kit. Immunocytochemistry analysis was designed to determine transforming growth factor beta1(TGF-β1) protein expression while immunofluorescence analysis for RAGE and NF-kB. The protein expressions of TGF-β1, RAGE and NF-kB were analyzed by Western blot analysis. Results Pretreatment with GA at a concentration of 10−8–10−6 M significantly reduced the AGEs-induced apoptosis in HUVECs. GA significantly increased antioxidant enzyme SOD activity and decreased peroxide degradation product MDA level in a dose-dependent manner. Furthermore, GA also remarkably inhibited the overgeneration of AGEs-induced ROS. Both immunocytochemistry analysis and western blot analysis showed that GA significantly decreased the protein expression of poinflammatory cytokine TGF-β1 in a similar manner which RAGE-Ab did. Additionally, AGEs-induced RAGE and NF-kB protein expressions were down-regulated significantly by the pretreatment with GA or RAGE-Ab. Conclusion These findings provide evidences that GA possesses protective activity on AGEs-induced endothelial dysfunction, including anti-apoptosis, anti-inflammation and antioxidant stress, via inhibiting RAGE/NF-kB pathway. GA might be an alternative for the prevention and treatment of diabetic vascular complications in an appropriate dosage.

Objective To study the impact of saikosaponin on absorption and transport of paeoniflorin in Caco-2 cell model. Method The concentration of paeoniflorin in cell culture medium was measured by UPLC and the apparent permeability coefficients (P(app)) was calculated to study differences in bi-direction transport of paeoniflorin solutions of different concentrations and its compatibility with saikosaponin a, and saikosaponin d in Caco-2 cell model. Meanwhile, the electric resistance of Caco-2 cell was determined before and after the experiment. Result The amount of paeoniflorin increased linearly with the transport of Caco-2 cell monolayer in 4 h, with a lower absorptive permeability, which was (0.98 +/- 0.10) x 10(-6), (0.92 +/- 0.09) x 10(-6), (0.89 +/- 0.04) x 10(-6) cm x s(-1) at the concentration of 20, 50, 100 micromol x L(-1), respectively. After compatibility with saikosaponin a and saikosaponin d, the absorptive permeability of paeoniflorin increased by 2.37 times and 2.54 times, respectively, while the electric resistance of Caco-2 cell was decreased significantly after the experiment. Conclusion Saikosaponin a, d can enhance the absorption of paeoniflorin in Caco-2 cell monolayer, which may be related to saikosaponin's ability to open up intercellular tight junctions among Caco-2 cells.

Cigarette smoking is the main cause of chronic obstructive pulmonary disease and lung cancer. The present study was aimed to explore the chemopreventive effect of ursolic acid (UA) on these diseases. In the CSE treated normal human bronchial epithelial cell model, UA alleviated cytotoxicity caused by CSE, recovered the intracellular redox balance, and relieved the stimulation of external deleterious factors as well. UA mitigated CSE-induced DNA damage through the Nrf2 (nuclear factor erythroid 2-related factor 2) pathway. Moreover, UA inhibited lung cancer development in the model established by A549 cells in nude mice in vivo. For the first time, our results indicate that UA could be developed as a potential lung cancer chemopreventive agent.

Application of fresh herbs is a kind of special forms of traditional Chinese medicine. In China, there is a long and rich experience in clinical application of fresh herbs. Many studies showed that the efficacy of fresh herbs was better than that of dried herbs, but the further study about the difference of their chemical composition, effective components and the overall material basis were few. In this paper, the ideas and methods to study on material basis of the fresh herbs by comparing the difference of the fresh and dry herbs in medicine chemical composition and pharmacological activity of effective components with modern advanced separation, analysis and screening technology under the "Constituent structure theory" were proposed. It was an effectual method for studying on the reasonable development of Chinese medicine and fresh herbs resources.

In this work, the metabolite profiles of icariin in rat plasma were qualitatively investigated, and the possible metabolic pathways of icariin were subsequently proposed. After oral administration of icariin, rat plasma samples were collected and pretreated by protein precipitation. Then these pretreated samples were injected into a Venusil ASB-C18 column with mobile phase consisted of 0.1% formic acid water and 0.1% formic acid acetonitrile and detected by Ultra fast liquid chromatography coupled to time-of-flight mass spectrometry (UFLC-TOF/MS). A total of 19 metabolites, namely, icariside I (M1), icaritin (M2), desmethylicaritin and its isomer (M3-M6), icaritin-3-O-gluA (M7), icaritin-7-O-gluA (M8), icariside II and its isomer (M9 and M10), icaritin-3,7-di-O-gluA (M11), 1,3-isoprene alcohol icaritin and its isomer (M12, M13 and M18), 1,3-isoprene alcohol icariside II (M14), allylic alcohol icaritin and its isomer (M15 and M16), 1,3-isoprene icariside II (M17) and icaritin-3-O-rha-7-O-gluA (M19) were detected and tentatively identified, and 9 of them, including M7 and M8 and M12-M18 were reported for the first time. The metabolites profiles in plasma revealed that glucuronide conjugates of isoflavonoids and flavonoid aglycones were the major circulating forms of icariin.

The main purpose of this study was to evaluate the intestinal absorption and the antineoplastic effect of the poorly water-soluble drug celastrol when liposomes were used as oral drug delivery system. Liposomes were prepared by the ethanol-injection method. An optimized liposome formulation composed of phospholipid, cholesterol and Tween-80 resulted in favorable encapsulation efficiency at 98.06 ± 0.94%. Homogeneous and stable particle size of 89.6 ± 7.3 nm and zeta potential of −(87.7 ± 5.8) mV were determined by laser particle size analyzer. Subsequently, the four-site perfusion rat intestinal model revealed that celastrol-loaded liposomes had improved effective permeability compared to the free drug in four intestinal segments (p < 0.05). Moreover, celastrol-loaded liposomes could also inhibit the tumor growth in C57BL/6 mice. These results suggest that liposomes could be a promising perioral carrier for celastrol.

Purpose: Yinyanghuo (Herba Epimdii) is a traditional Chinese herb containing prenylated flavonoids as its active constituents. The aim of this study was to examine the significance of the intestinal hydrolysis of prenylated flavonoids by lactase phlorizin hydrolase (LPH), an enzyme at the brush border membrane of intestinal cells. Methods: A four-site perfused rat intestinal model was used. The concentration of the flavonoids of interest and their metabolites in different intestinal segements were analyzed by HPLC, and the apparent permeabilities were calculated. A lactase phlorizin hydrolase inhibitor (gluconolactone) was employed to investigate the mechanism of the intestinal absorption, and the metabolites of the four flavonoids were identified using LC/MS/MS. Results: Diglycosides (icariin) or triglycosides (epimedin A, epimedin B, and epimedin C) were hydrolyzed rapidly in duodenum and jejunum producing one or two metabolites, while a monoglycoside (baohuoside I) was absorbed directly. When co-perfused with glucono-lactone, both the hydrolysis of diglycosides and triglycosides were significantly inhibited, with inhibition rates for icariin (62%, 50%, 40%, 46%), epimedin A, (55%, 26%, 21%, 14%); epimedin B (42%, 40%, 74%, 22%), and epimedin C (42%, 40%, 52%, 35%) in duodenum, jejunum, ileum, and colon, respectively. Also the metabolites of icariin, epimedin A, epimedin B, and epimedin C were identified as baohuoside I (one of two), sagittatoside A, sagittatoside B, and 2"-O-rhamnosylicariside II, respectively. Conclusions: The results showed that lactase phlorizin hydrolase was a major determinant of the intestinal absorption of prenylated flavonoids present in Yinyanghuo.

The efficacy of Prunella extracts in the prevention and treatment of lung cancer has been attributed to different components. In this study, an "active components combination model" hypothesis was proposed to explain the anti-tumor activity of Prunella. The efficacy of Prunella extracts from different regions was compared in vitro and in vivo, and the TNF-α activity in serum of tumor-bearing mice was also evaluated. High performance liquid chromatography (HPLC) was used to analyze the extracts and identify 26 common peaks. Prunella samples from different regions were classified by the cluster analysis method; both P. vulgaris L. from Bozhou and P. asiatica Nakai from Nanjing, which had the highest activities, were further divided into different classes. Six peaks from the HPLC analysis were very similar, and were identified as caffeic acid, rosmarinic acid, rutin, quercetin, oleanolic acid and ursolic acid. The total ratio of these compounds in Prunella from Bozhou and Nanjing were 1.0:14.7:3.9:1.0:4.4:1.4 and 1.0:14.8:4.0:0.8:5.6:1.8, respectively. Total triterpenes and total phenols in Prunella were separated by macroporous resin purification for activity studies. The results showed that total triterpenes and total phenols had anti-lung cancer activity and their combination significantly enhanced the activity. In addition, the combination also significantly increased the TNF-α content compared to total triterpenes or total phenols. The results indicated that the efficacy of Prunella against lung cancer was attributable to multiple components acting at an optimal ratio.

Objective To investigate the intestinal absorption of naringin, hesperidin, neohesperidin and the extract of Fructus Aurantii Immaturus in rats. Method The rat intestinal perfusion and enzymes incubation models were used, together with the determination of the n-octanol/water partition coefficient of the components (P). Result In perfusion model, the P(eff) of all components were low, and the P(eff) of naringin, hesperidin and neohesperidin were 0.140-0.252, 0.156-0.268 and 0.154-0.285, respectively. In four different regions of intestine of rat and with different concentration, the P(eff) of the components both had no significant difference, whereas the P(eff) of the extract were higher than the P(eff) of the single component. The metabolite of components was not detected in intestine. The P of naringin, hesperidin and neohesperidin were 0.36, 0.40 and 0.48, respectively, and the pH of buffer solution had no influence to its distribution coefficient. Conclusion Poor permeation contributed to the poor intestinal absorption of naringin, hesperidin and neohesperidin. The absorption of components in extract were increased, and the results suggest that the extract may enhance the intestinal absorption of the components.

OBJECTIVE To prepare CIT microemulsion and to investigate its properities and the absorption character in rat intestine in situ. METHOD The optimum formulation of the blank microemulsion was selected by pseudo tertiary phase diagrams and the CIT microemulsion was prepared based on the blank microemulsion. The appearance, particle diameter distribution and Zeta potential were investigated by electron microscopy and grainsize analyzer. The entrapment efficiency was determined by sephadex column chromatography. An in situ single pass intestine perfusion method was used to investigate the intestinal absorption of CIT microemulsion. HPLC method for determination of CIT in the intestinal flux was established. RESULT The formulation was OP-glycerol-water-ethylis oleas (3:3:4:2). CIT microemulsion in electron microscopy was consisted of small spherical drop. The mean diameter was 61.7 nm, Zeta potential was -46.3 mV. The Entrapment efficiency was (92.1 +/- 1.74)% (n=3). Both CIT and CIT microemulsion had been absorbed in general intestinal tract,and the absorption of CIT microemulsion was significantly high compared with CIT in duodenum, colon, jejunum and ileum (P < 0.01). CONCLUSION The preparation technology of CIT microemulsion was feasible,and the microemulsion system might improve the absorption of CIT in the intestinal tract.

Background: Ginseng, the root of Panax ginseng (PG), is used widely as a herbal medicine to prevent and treat various diseases. Panax ginseng has pharmacological effects on neurodegenerative diseases such as Alzheimer's disease (AD). The present study evaluated the neuroprotective effects of PG and its possible neuroprotective mechanisms in advanced glycation end product (AGE)-induced AD in a rat model. Methods: Advanced glycation end products were injected bilaterally into the CA3 region of the rats' brains. The Morris water maze test and step-down type passive avoidance test were performed to evaluate their memory and cognitive abilities. The oxidation indexes in the hippocampus were detected. Immunohistochemistry was conducted to visualize the receptors for advanced glycation end products (RAGEs) and nuclear factor-kappa-light-chain-enhancer of activated B cell (NF-κB). Results: Behavioral results showed that PG (1 g/kg, 0.5 g/kg, and 0.25 g/kg) significantly shortened the escape latency, remarkably increased the number of crossing times, significantly decreased the number of errors, and prolonged the latency in rats with AGE-induced AD. Panax ginseng also significantly reduced the malondialdehyde level, increased the glutathione content, and increased superoxide dismutase activity in the hippocampus. Panax ginseng significantly decreased the expression of RAGE and NF-κB. The blockade of anti-RAGE antibody could significantly reduce AGE-induced impairments and regulate these expressions. Conclusion: Our results demonstrated that PG significantly inhibits AGE-induced memory impairment and attenuates Alzheimer-like pathophysiological changes. These neuroprotective effects of PG may be associated with the RAGE/NF-κB pathway. Our results provided the experimental basis for applying PG in preventing and treating AD.