Diallyl Disulfide Affect Extracellular Matrix Synthesis And TGF-β1/Smad Signaling Pathway In Rat Hepatic Stellate Cells

Objective To explore the effects of diallyl disulfide (DADS) on extracellularmatrix synthesis and TGF-β1/Smad signaling pathway in rat hepatic stellate cells.Methods (1) Activated rat hepatic stellate cells HSC-T6were randomly andaveragely divided into five groups,vehicle control group (A), treatment groups weregiven DADS in different concentrations（B20mg/L;C40mg/L;D80mg/L),treatment group was given2.5×10-7M colchicine（E）;(2) Detect the expression ofcollagen typeⅠ(ColⅠ) and hyaluronic acid (HA) by Enzyme-linked immunosorbentassay (Elisa).(3) Detect the transcription and expression of TGF-β1/Smadsignaling pathways associated protein TGF-β1, Smad3, pSmad3and Smad7by RT-PCR,Western blot.The results (1) DADS can inhibit the Synthesis of extracellular matrix in rathepatic stellate cells, reduce the synthesis of collagenⅠ(ColⅠ) and HA (hyaluronicacid) in a dose-dependent manner. Elisa results show that DADS (20mg/L,40mg/L,80mg/L) and Colchicine effect respectively in the HSC-T6cells after48h，compared with the vehicle control group, the synthesis of extracellular matri of thetreatment groups was significantly reduced (P <0.05), the difference was significant;(2) RT-PCR results show that compared with the vehicle control group,the expressionof TGF-β1mRNA of40mg/L group,80mg/L group, colchicine group wassignificantly reduced,the difference was significant.(P <0.05), the most obviousdecrease was80mg/L group,20mg/L group was no significant difference; Theexpression of Smad3mRNA between the groups had no statistically significant (P>0.05).(3) Western blot result show that compared with the vehicle control group, theexpressions of TGF-β1and pSmad3protein of40mg/L group,80mg/L group,colchicine group were significantly reduced, the difference was statisticallysignificant (P <0.05), and the expressions of TGF-β1protein and pSmad3protein of DADS intervention groups were in a dose-dependent manner,80mg/L group wasthe most obvious,20mg/L was no statistically significant difference (P>0.05); Theexpressions of Smad3protein in each group were no statistically significant (P>0.05);Compared with the vehicle control group,the expressions of Smad7protein of40mg/L,80mg/L, colchicine group were significantly increased,the difference wasstatistically (P <0.05), the expressions of Smad7protein of DADS intervention groupswere in a dose-dependent manner,80mg/L group was the most obvious,20mg/Lwas no statistically significant difference (P>0.05).Conclusions In this experiment, DADS inhibited the synthesis of extracellularmatrix of HSC-T6cells in a dose-dependent manner, the possible mechanism wasthat by affecting the TGF-β1/Smad pathway, reduced the expression of TGF-β1,increased the expression of Smad7, inhibited the phosphorylation of Smad3.