Relation of TGF-β3/TGF-β1 mRNA ratio to expression of TGF-β1,MMP-9 and TIMP-1 in rat hepatic stellate cells

Abstract

Objective To investigate the change of TGF-β3/TGF-β1 mRNA ratio and explore its correlation with TGF-β1,MMP-9,TIMP-1 expression in rat hepatic stellate cells.Methods Plasmids pcDNA3.1 (+)- TGF-β3 and pcDNA3.1 (+)-TGF-β1 were constructed,pcDNA3.1 (+)-TGF-β1 was transfected into HSC-T6 cell line and the positive cell clone with high expression of TGF-β1 was established after being filtrated,pcDNA3.1 (+)- TGF-β3 was transfected into the positive clone.Fortyeight hours later,the mRNA and the protein expression of TGF-β3,TGF-β1,MMP-9 and TIMP-1 was detected with real-time PCR and Western blot.Results TGF-β3/TGF-β1 mRNA ratios in the blank group,the control group,the positive clone group and the TGF-β3 intervene group were 0.286+0.070,0.874±0.141,0.448±0.327 and 277±0.244,respectively.The expression of TGF-β1 and TIMP-lmRNA and protein were much higher in the positive clone group than those in the blank group and the control group(P<0.05).The expression of MMP-9 mRNA and protein was much lower in the positive clone group than those in the blank group and the control group(P<0.05).Comparing the TGF-β3 intervene group with the positive clone group,TGF-β1 protein and TIMP-1 mRNA and protein decreased significantly(P<0.05).MMP-9 mRNA and protein increased significantly (P<0.05).Conclusion TGF-β3 can decrease the expression of TGF-β1 protein.More expression of TGF-β3 than that of TGF-β1 can inhibit the synthesis of TGF-β1 and TIMP-1,and encourage the expression of MMP-9 in HSC.When TGF-β3/TGF-β1 mRNA ratio is less than 1,the expression of TGF-β1 protein decreases,but there are no significant changes in MMP-9 and TIMP-1. Key words: Hepatic stellate cell; TGF-β3/TGF-β1 mRNA ratio; Matrix metalloproteinase-9; Tissue inhibitor of metalloproteinase-1