Effects of recombinant transforming growth factor-β_3 gene on protein expressions intracellular and extracellular of hepatic stellate cell

Objective To explore the influence of recombinant transforming growth factor-β3(TGF-β3) on protein synthesis and secretion intracellular and extracellular of hepatic stellate cell(HSC).Methods Plasmid pcDNA3.1(+)-TGF-β3 and pcDNA3.1(+)-TGF-β1 were well constructed.A positive cell clone stably and highly expressing TGF-β1 was established after being screened by G418 medium,and the positive clones were transfected by recombinant TGF-β3.HSC-T6 cells were cultured and divided into four groups: blank control group;pcDNA3.1(+)-enhanced green fluorescent protein(EGFP)-transfected group(negative control group);pcDNA3.1(+)-TGF-β1 transfected group(TGF-β1 positive clone group);pcDNA3.1(+)-TGF-β3 transfected positive clones group(TGF-β3 interfered group).Western blot and ELISA were respectively used to detect the protein synthesis and secretion of TGF-β1,typeⅠ collagen,matrix metalloproteinase(MMP)-2,MMP-9 and tissue inhibitor of metalloproteinase(TIMP)-1.Results The protein expressions of TGF-β1,type Ⅰ collagen,MMP-2 and TIMP-1 in the TGF-β1 positive clone group were obviously higher than those in the blank control group and negative control group(P0.05),while the protein expression of MMP-9 was significantly lower.Compared with the TGF-β1 positive clone group,the protein expressions of TGF-β1,type Ⅰcollagen and TIMP-1 in TGF-β3 interfered group were much lower(P0.05),and MMP-2 was lower,but the difference had no statistical significance.The protein expression of MMP-9 was obviously higher(P0.05).Conclusion Recombinant TGF-β3 reduces the synthesis and secretion of type Ⅰ collagen,and inhibits the collagen deposition by adjusting the expression of matrix metalloproteinases and their inhibitors.