Effects Of Tetramethylpyrazine On Expression Of CTGF,Smad2/3 In Hepatic Stellate Cells

Objective: To study the effects of Tetramethylpyrazine(TMP) on expression of connective tissue growth factor(CTGF) and Smad2/3 in hepatic stellate cell-T6 (HSC-T6).Methods:HSC-T6 was cultured with varying doses of TMP and 0.005mg/l[1] of transforming growth factor-β1(TGF-β1) in vitro.MTT colorimetric assay was used for detecting the proliferation inhibition ratio of HSC-T6.Immunocytochemistry was used to observe the location of CTGF and Smad2/3 ,to test the effect of TMP on the expression of CTGF and Smad2/3 in HSC-T6.Protein expression level of CTGF and Smad2/3 was analyzed with Western blot.Results:1. TMP inhibited the proliferation of HSC-T6 at the concentration of 100,200,400,600 and 1000mg/l in concentration dependent way(P<0.05).2. The test by immunohistochemical method indicated that the expression of CTGF located in the endochylema of HSC-T6 while Smad2/3 expressed both in the nucleus and endochylema of HSC-T6. When the density of TMP increased, the expression of CTGF and Smad2/3 decreased progressively. The comparision between the control group were remarkably discrepant (P<0.05).3. The results of Western blot also revealed that TMP could dose- dependently inhibit the protein expression of CTGF and Smad2/3,the inhibition drgree of which showed a positive correlation.(r=0.967,P<0.01).Conclusion:Inhibiting proliferation of HSC-T6, down-regulating the protein expression of Smad2/3 and CTGF,might be involved in the anti-fibrosis mechanism of TMP.