The Effect Of Ganfukang On Fas/caspase8Pathway In Hepatic Fibrosis

Objective: Observe the expression of Fas/caspase8pathway in experimentalhepatic fibrosis and the effect of traditional Chinese medicine-Ganfukang on itsexpression, from the molecular mechanism to explore Fas/caspase8pathway in thetreatment of hepatic fibrosis in Ganfukang. For the treatment of hepatic fibrosis toprovide a new theoretical basis.Methods: Healthy rats were randomly divided into normal group (N), modelgroup (M), high-dose-group (HD), middle-dose-group (MD), low-dose-group (LD). Inaddition to the normal group injected with normal saline, the concentration ofsubcutaneous injection of olive oil diluted carbon tetrachloride twice a week, a total of12weeks. High-dose-group, middle-dose-group,low-dose-group in the model to9weeks,after gavages to3125mg/kg/d and312.5mg/kg/d,31.25mg/kg/d dose of Gan-fu-kang12weeks of treatment. Normal control group and model group while the same doseof normal saline treatment. Normal group and model group were sacrificed on the12thweek,20weekend the same way the other groups were sacrificed. In vitro culture, giveacetaldehyde intervention of HSC-T6cells were divided into normal group (N), modelgroup (M) and medication group (middle-dose, MD). The normal control group(N):inthe10％normal control groups in DMEN medium, incubated for48hours;acetaldehyde stimulation group (M): in the10％normal control group serum DMEMculture medium added with acetaldehyde, cultured for48hours; drug therapy group(MD): with10％doses in pharmacology. DMEM culture medium added withacetaldehyde, cultured for48hours.(Acetaldehyde final concentration of200μmol)HE staining method for the detection and observation of rat liver tissuepathological changes; immunohistochemistry SP method for the detection of Fas proteinin liver tissue of each group; RT-PCR method for the detection of Fas, FADD, caspaes8,P53in liver tissue of each group and the expression of mRNA in cells; the protein expression of Fas in liver tissue and cells of each group were detected by Western Blot.Results:1. The histopathological changes of rat liver tissue: HE staining revealed that,compared with the normal group, the structure of hepatic tissue in model groupdisordered and chaos, hepatic lobules were destroyed or even disappear, per portalappeared thick fibrous cord. Compared with model group, middle dose groupsignificantly improved the above changes.2. Fas protein in the liver tissue of expression was significantly higher than that innormal group, while the middle dose group compared to the model of Fas proteinexpression was significantly reduced.3. Fas pathway and P53mRNA expression in the target tissue and cell expressionshowed the same trend. Model group, the expression of Fas pathway and P53mRNAsignificantly higher than the normal group, model group compared to Fas pathways,middle dose group significantly decreased, while the P53dose was continued to rise.4. Western Blot assay result, Fas protein expression in cells of the same trends inthe organization. Middle dose group significantly decreased compared with the modelgroup.Conclusion:1. Fas/caspase8pathway rose to promote action in the development of hepaticfibrosis.2. Chinese medicine-Ganfukang has a therapeutic effect on hepatic fibrosis, andits mechanism may be related to Fas/caspase8mediated death receptor pathway.