Effects Of Resistin On Hepatic Fibrosis Possible Mechanism In Non-alcoholic Fatty Liver Disease In Invtro And In Vivo

ObjectiveTo investigate the resistance factors for nonalcoholic fatty liverdisease (non-alcoholic fatty liver disease. NAFLD) hepatic fibrosis in ratsand possible vivo and in vitro mechanisms.MethodsThe establishment of the NAFLD liver fibrosis in vivo model toapplication of high fat diet,30wistar rats were randomly divided intocontrol group and model group,15were fed a normal diet and high fat diet,respectively, were sacrificed10at15,18,21weeks (control group, modelgroup5). Hematoxylin-eosin staining (HE staining) and collagen fiberstaining (VG staining) observed in liver tissue fibrosis, the electronmicroscope the HSC (hepatic stellate cells) activation and liver fibrosis;Determination of serum the level of fibrosis index of procollagen type Ⅲ(PC ⅲ), hyaluronic acid (HA), collagen type IV (CIV), laminin protein(LN), using semi-quantitative RT-PCR and by Western blot analysis of ratliver tissue resistin expression of the trend observed between resistin andhepatic fibrosis. Selection of recombinant resistin role NAFLD liver fibrosis in vitro model of HSC-T6. Cultured with RPMI-1640mediumcontaining10%fetal bovine serum in HSC-T6cells covered the culturebottle after75%passage upon adherent divided into4groups: group A(1640culture medium); B, C, D, respectively, at different concentrationsagainst hormone25ug/L,50μg/L, and100ug/L, the cells were culturedfor72hours, cells were detected in the supernatant procollagen type Ⅲ(PCⅲ) by1640, hyaluronic acid (HA) in type Ⅳcollagen (CIV), lamininprotein (LN) level; using semi-quantitative by RT-PCR, method to detectthe HSC-T6, in transforming growth factor beta-1(of TGF-beta1) mRNAin and tumor necrosis factor alpha (TNF-alpha) mRNA expression levels.Analysis of resistance mechanism of action of the hormone on the HSCResultsWith the high fat diet rats extension of time, the liver tissue fibrosischanges in serum hepatic fibrosis markers procollagen type Ⅲ (PC Ⅲ),hyaluronic acid (HA), type IV collagen (CIV), laminin (LN) and the levelgradually increased, indicating that fibrosis gradually increased; theactivation of HSC in the liver tissue resistin expression gradually increased.HSC-T6supernatant fibrosis index III procollagen (PC III), hyaluronic acid(HA), collagen type IV (C IV), laminin (LN) level gradually increased withthe resistance to the increase of hormone concentration, that fibrosisgradually increased; increased expression of TGF-β1mRNA and TNF-αmRNA in the cells. Compared with the control group, the differences in each indicator are significant (p <0.05or0.01)..ConclusionsAnimal models, expression of resistin increase in liver tissue,activation of HSC and liver fibrosis increased in the serum, liver fibrosislevels gradually increased, but the causal relationship between resistin andfibrosis is unclear.Therefore, to study the in vitro cell model, studies haveshown that resistin gradually increased to promote the level of liver fibrosisand TNF-α mRNA expression of TGF-β1increase, suggesting thatresistance factors may be the role in the HSC to produce TNF-alpha andTGF-β1and other inflammatory factors induce the occurrence of liverfibrosis in NAFLD development.