Interventional effect of asiaticosdide on rats exposed to silica dust / 中华劳动卫生职业病杂志

Objective: To investigate the effect of asiaticoside for fibrosis in lung tissues of rats exposed to silica and to explore its possible mechanism. Methods: 144 SD male rats were randomly divided into control group, model group, positive drug control group, asiaticoside high-dose group, medium-dose group and low-dose group, each group included 24 rats. Rats in the control group were perfused with 1.0 ml of normal saline, and the other groups were given 1.0 ml 50 mg/ml SiO(2) suspension. Gavage of herbal was given from the next day after model establishment, once a day. Rats in the positive drug control group were administration with 30 mg/kg tetrandrine and rats in the low-dose group, medium-dose group and high-dose group were given 20 mg/kg, 40 mg/kg and 60 mg/kg asiaticoside for fibrosis respectively. Rats in the control group and the model group were given 0.9% normal saline. The rats were sacrificed in on the 14th, 28th and 56th day after intragastric administration and collect the lung tissues to detect the content of hydroxyproline, TGF-β(1) and IL-18, observe the pathological changes of the lung tissues by HE and Masson staining and determine the expressions of Col-I, a-SMA, TGF-β in lung tissues by Western Blot. Results: On the 14th day, 28th day and 56th day after model establishment, the lung tissues of rats in the model group showed obvious inflammatory response and accumulation of collagen fibers, and the degree of inflammation and fibrosis increased with time. The intervention of asiaticoside could effectively inhibit the pathological changes of lung tissues. The contents of hydroxyproline, IL-18 and TGF-β1 in lung tissues of model group were higher than those in the control group (P<0.05) , while the level of hydroxyproline, IL-18 and TGF-β1 in asiaticoside groups were significantly decreased, and the difference was statistically signicant (P<0.05) . Compared with the control group, the expression levels of Col-I, TGF-β1and α-SMA in lung tissue of model group were increased (P<0.05) , while the expression level of Col-I, TGF-β1 and α-SMA were decreased after the intervention of asiaticoside, and the difference was statistically signicant (P<0.05) . Conclusion: Asiaticoside can inhibit the increase of Col-I, TGF-β1 and α-SMA content in the SiO(2)-induced lung tissues of rats, reduce the release of TGF-β1 and IL-18 inflammatory factors in lung tissue, and then inhibit the synthesis and deposition of extracellular matrix in rat lung tissue, and improve silicosis fibrosis.

Effect of Tetrandrine on Col-I and FN in TGF-β1-induced MRC-5 Cells / 中国实验方剂学杂志

Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.

Effect and mechanism of tanshinone IIA on TGF-β1 induced human skin fibroblast proliferation / 中草药

Objective: To study the effect and mechanism of tanshinone IIA on human skin fibroblasts cell (HSF). Methods: CCK- 8 method was used to determine the effect of different concentrations of TSA on the proliferation of HSF induced by TGF-β1. The plate cloning ability of HSF treated with different concentrations of TSA (2.5, 5, 10 and 20 μmol/L) for 48 h were analyzed by plate clonogenesis assay. The protein expression of TGF-β1/Smad signaling pathway proteins and α-SMA, VEGFA and COL I were further measured by Western blotting. Results: CCK-8 and plate clonognesis assay results showed that TSA significantly inhibited the proliferation and colony forming efficiency of HSF (P < 0.01). Western blotting results revealed that each concentration group of TSA significantly inhibited the protein levels of p-Smad2 and p-Smad3, and down-regulated the ratio of p-Smad2/Smad2 (P < 0.01). The ratio of p-Smad3/Smad3 was significantly decreased in 5, 10 and 20 μmol/L TSA groups. Additionally, the expression levels of α-SMA, VEGFA and COL I in HSF decreased significantly with the increase of TSA concentration (P < 0.01). Conclusion: TSA exhibits the inhibitory effect on proliferation of HSF, and its mechanism may be related to TGF-β1/Smads signaling pathway.

Preliminary study of resveratrol on anti -hepatitis B virus and hepatitis induced fibrosis / 中国生化药物杂志

Objective To study the effect of resveratrol on hepatitis B virus (HBV) and hepatic fibrosis.Methods The rat model of HBV infection was established by intraperitoneal injection of rAAV8-1.3HBV.After successfully establishing the modle, were randomly divided into four groups:negative control group, resveratrol low dose group, high dose group, positive control group, and five rats in each group.Giving a physiological saline, 20 mg/kg respectively resveratrol, 40 mg/kg, 0.1 mg/kg acyclovir via gastrogavage respectively.2、4、8、12 weeks later ,we detected the HBsAg, HBeAg, HBV-DNA via tail vein blooding.HSC-T6 cell model was established.Then rats were divided into two groups, control and resveratrol serum group.Each group was administrated with normal sodium and resveratrol via gastrogavage to make serum.The HSC-T6 viability was observed by AlamarBlue assay.The expression of Col I and TGP-β1 mRNA was determined by Realtime PCR.The expressions of Col I and TGP-β1 Protein was analyzed by WesternBlot.Results Using ELISA method to detect HBsAg、HBeAg、HBV-DNA and found that the expression of HBsAg and HBeAg of negative control group were significantly higher than the rest of the three groups (P<0.05) at the second and the fourth week.The highest expression of HBsAg and HBeAg were (4118 ±367) IU/mL、(160.2 ±56.90) NCU/mL at the second week.The expression of HBV-DNA of negative control group is significantly higher than the rest of the three groups (P<0.05) at the second week.AlamarBlue assay indicated that different concentration of serum of resveratrol can inhibit HSC-T6 proliferation.Compared with control group, serum with drug resveratrol significantly down-regulated the mRNA and protein expression of Col I and TGP-β1.Conclusion Resveratrol inhibits HBV and liver fibrosis by inhibiting type I collagen and TGF-β1.

Analysis on clinical infection distribution and drug resistance of 442 strains of Escherichia coli / 国际检验医学杂志

Objective To analyze the clinical infection distribution and drug resistance status of 442 strains of Escherichia coli to provide the basise for the treatment of Escherichia coli infection and the control of nosocomial infection.Methods The clinically submitted various kinds of specimens during 2013 were performed the bacterial culture and identification.The susceptibility of Escherichia coli to commonly used bacterial drugs were detected by adopting the MIC method.The data were analyzed by WHO-NET V5.5 and SPSS V13.0 softwares.Results 442 strains of Escherichia coli were isolated from the middle urine and secretion. The detection rate of ESBLs-producing Escherichia coli was 61.3%.442 strains of Escherichia coli had the high resistance to peni-cillins,cephalosporins and fluoroquinolones,better sensitivity toβ-lactam/β-lactamase inhibitor compounds and highest sensitivity to carbapenems.ESBLs-producing Escherichia coli had the higher resistance to commonly used antibacterial drugs than non-ESBLs-producing Escherichia coli .Conclusion The drug resistance of Escherichia coli is severe.ESBLs-producing Escherichia coli are u-sually resistant to many different types of antimicrobial drugs.Carbapenems are the first choice to treatment of severe infections of Escherichia coli .

Effects of halofuginone on hepatic fibrosis in rats and their mechanism / 中草药

Objective: To investagate the protective effects of halofuginone against liver injury induced by concanavalin A (Con A) in rats and their mechanism. Methods: Rats were divided into control (G1, 8), model (G2, 14), low- (5 mg/kg, G3, 14) and high-dose (10 mg/kg, G4, 14) halofuginone groups. The rats in G1 were iv injected with 300 μL PBS from tail vain once a week, the rats in other three groups were iv injected with 12.5 mg/kg Con A (in 300 μL PBS) once a week for eight consecutive weeks. Halofuginone was given in the diet for rats in G3 and G4 after molding and continuous administration for eight weeks after the rats were weighted and then were put to execution. The biochemical analysis was used to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), and album protain (ALB) in serum while enzyme-linked immuno sorbent assay (ELISA) was applied for detecting transforming growth factor β1 (TGF-β1), hyaluronic acid (HA), and procollagen III (PC-III) levels in serum. HE and Masson's trichrome stainings were conducted in liver tissue to observe the pathological variations. Grades of hepatic fibrosis were evaluated according to SSS system. Sirus and immunohistochemical stainings were executed for detecting the deposition and protein expression of collagen 1 (Col I) in liver tissue. Results: Compared with the G2, halofuginone could decrease the levels of ALT, AST, TP, ALB, TGF-β1, HA, and PC-III in serum obviously (P < 0.05, 0.01). Halofuginone could improve the liver pathological variations of fibrotic rats obviously (P < 0.05, 0.01) and reduce the score of hepatic fibrosis significantly (P < 0.05, 0.01). Halofuginone treatment could reduce the deposition and expression of Col I protein (P < 0.05, 0.01). Conclusion: Halofuginone could attenuate the Con A-induced immunological liver injury and the fibrosis level in rats. The mechanisms possibly contribute to down-regulating the deposition and expression of Col I protein in liver of rats.