Effects Of Tetramethylpyrazine And Silencing TGF-B1by Sirna On Expression Of Connective Tissue Growth Factor Of Rat Hepatic Stellate Cells

OBJECITVE:To investigate the effect of TMP and chemosynthesis siRNA TGF-β1on the expression of CTGF of rat hepatic stellate cells(HSC), which provide theory foundation to investigate a new way of anti-hepatic fibrosis.METHODS:HSC were cultured in vitro,screening out a effective segment of gene silencing from three segments of siRNA TGF-β1,then it was transfection group (S) utilizing lipsome transfection reagent to transfect together the cells which have been cultured for24h,and designing blank control group(B),transfection reagent group(R), TMP group(T),and union treatment group (S+T).We collected HSC and culture supernatant after incubation for72h.The expression of type Ⅰ,Ⅲ collagen and CTGF mRNA were detected by RT-PCR., The expression of CTGF protein in HSC was detected by Western blot, Contents of type Ⅰ and Ⅲ collagen in supernatant were determined by ELISA.RESULTS:1.Compared with blank control group, the test by RT-PCR method indicated that the expression of Ⅰ,Ⅲ collagen and CTGF mRNA were reduced in transfection group by(19±3%、32±2%、14±4%, p<0.05-0.01)respectively,in TMP group by(30±2%、51±2%、25±3%, p<0.05-0.01)respectively,and in union treatment group by(37±5%、56±3%、36±3%,p<0.05-0.01)respectively.2.Compared with blank control group,the results of Western blot revealed the CTGF protein expression was down-regulated by(16.2±3%、25.2±3%、41.6±4%,p<0.05-0.01)in transfection group,TMP group and union treatment group respectively.3.Compared with blank control group,the test by ELISA indicate contents of type Ⅰ collagen in supernatant was reduced by (6.1±5%、12±6%、15±5%,p<0.05-0.01)respectively,type Ⅲ collagen was reduced by(13±3%、24±4%、41±2%,p<0.05-0.01)respectively.CONCLUSION:The expression of CTGF can be down-regulated by silencing TGF-β1by siRNA,which can reduce the expression of type Ⅰ and Ⅲ collagen. The expression of CTGF can be also down-regulated by TMP,but it is more pronounced to the inhibiting expression of type Ⅰ and Ⅲ collagen,cluing that anti-hepatic fibrosis of TMP may be multifarious action targets.