The Clinical Research On Treatment Of Liver Fibrosis And The Experimental Research On The Effects Of TGF-β1 Signal Transduction With Drug "Penthorum Chinense"

【Aim】To investigate the therapeutic effect of drug "Penthorum chinense" on treating liver fibrosis and Effects of drug "Penthorum chinense" on TGF-β1 activated the signal transduction exterior and interior of hepatic stellate cell.【methods】一,Clinical Research206 patients with liver fibrosis due to chronic hepatitis B were randomly divided into two groups,the control group(n=100)were treated by some routine treatment:in the first month treated with Diammonium Glycyrrhizinate 30ml and Tiopronin 0.3g,once a day;in second and third month treated with orally wuzhi gelatin capsule,two pills each time, three times per day;the therapeutic group(n=106)as the control group plus Gansu granule,one sack each time,three times per day;All patients were treated for three months,After treatment the clinical symptoms, syndromes,liver function,and PGA,3 indexes of liver fibrosis(HA,LN, C-Ⅳ),and the haemodynamics of liver were observed.二,Experimental ResearchHSC-T6 cells were divided into3 groups,normal group,model group,and″Penthorum chinense Extractive″group,which were cultivanted with DMEM including 0.5%NBS.the model group were simultaneously added into TGF-β1,while the″Penthorum chinense Extractive″group were simultaneously added into different concentration of drug and TGF-β1. The HSC-T6 viability was observed by AlamarBlue assay,while the toxicity of″Penthorum chinense Extractive″was measured by MTT assay. According to the result of the AlamarBlue assay and MTT assay,″Penthorum chinense Extractive″group with 2mg/ml was cultivanted with DMEM including 0.5%FBS and TGF-β1.After 24h,The expression of ColⅠ,α-SMA,ERK protein and phosphorylation,Smads protein and phosphorylation were analyzed by Western Blotting.【Results】1,Clinical ResearchAfter a one-month treatment,the clinical symptoms and syndromes were obviously improved in both two groups,77.4%of the therapeutic group patients' hepatitis conditions were disappear while 40%in control group. There was significantly different in the synthetically curative affection of the two groups(p＜0.01).After a three-month treatment,the indexes of liver function of two groups were both decreased obviously.There was significantly difference before and after treatment in those indexes of the therapeutic group (P＜0.01),as well as in control group(P＜0.05).After treatment,the decrease of ALT,AST,γ-GGT,TB of therapeutic group was significantly lower than the control group(P＜0.01).After treatment,as to the figures of HA,LN,IV-C,There was significantly difference before and after treatment in those indexes of the therapeutic group(P＜0.01),The figure of HA in the control group was also significantly lower than before(P＜0.05)while There was no significantly difference in the The figure of LN,IV-C in the control group. Compared with the control group,the figure of HA in the therapeutic group was significantly lower(P＜0.01)while those of LN,LN,IV-C were significantly lower(P＜0.05).After treatment,There was significantly difference before and after treatment in the diameter of portal and splenic vein of the therapeutic group(P＜0.01).compared to the control group,there was significantly difference after treatment in the diameter of portal and splenic vein of the therapeutic group(P＜0.01).While There was no significantly difference in the diameter of portal and splenic vein of in the control group(P＞0.05).There was no significantly difference in the spleen width in both two groups(P＞0.05)After treatment,the PGA parameter in two groups were both reduced. There was significantly difference before and after treatment in PGA parameter of the therapeutic group(P＜0.01).compared to the control group, there was significantly difference after treatment in PGA parameter of the therapeutic group(P＜0.05).While there was no significantly difference before and after treatment in PGA parameter of in the control group(P＞0.05).2,Experimental ResearchDifferenent concentration of drug″Penthorum chinense Extractive″can inhibit HSC-T6 proliferation,which wre the most manifest at 24h and in 2mg/ml concentration of drug″Penthorum chinense Extractive″;MTT assay indicated that drug″Penthorum chinense Extractive″extractum with 2mg/ml have unapparente toxicity to the cell,while drug″Penthorum chinense Extractive″with 10mg/ml and 50mg/ml have apparente toxicity,drug″Penthorum chinense Extractive″significantly down-regulated protein expression of ColⅠ,α-SMA,compared to model group,there was significantly difference after treatment in the drug″Penthorum chinense Extractive″group(P＜0.01);there were no obvious changes in protein expression of ERK and Smad2,but phosphorylation of ERK and Smad2 protein expression was up-regulated significantly,compared to model group,there was significantly difference after treatment in the drug″Penthorum chinense Extractive″group(P＜0.01).【Conclusion】″Penthorum chinense Extractive″could effectively improve the clinical symptoms,decrease the transaminase,resolve jaundice and promote liver functional recovery.Furthermore,it was effective in intercepting or reversing liver fibrosis proceeding,It couldn't effectively improve spleen width;Its mechanism may be related to relieve the liver inflammation and reduce the generation of TGFβ1 to inhibit the invigoration and activation maintaining of HSC;to reduce the generation and accumulation of collagen;to reduce generation and accumulation of extracellular matrix and encourage degradation of collagen. drug″Penthorum chinense Extractive″significantly decreased a-SMA and ColⅠsecretion at activated hepatic stellate cells,inhibit of the deposition of ECM in liver,its part mechanism of action may be concerned with that inhibit the ERK and Smads signal transduction of TGF-β1.In short,Drug″Penthorum chinense has better efficiency in the treatment of patients with liver Fibrosis.It was necessary of further research and application.