Effects Of Total Flavonoids Of Lichi On Expression Of MMP2 And TIMP1 In HSC-T6 Rat Hepatic Stellate Cells

Objective: To explore the effect of total flavonoids of lichi(TFL)on rat hepatic stellate cells(HSC-T6)cell proliferation,the influence of tissue inhibitor of metallo-proteinase 1(TIMP1)and matrix metalloproteinase 2(MMP2)gene and protein expression.To study the mechanism of action of TFL against hepatic fibrosis.Methods : Cultured rat hepatic stellate cells in vitro and divided into four grouped as follow: TFL high dose group(including 640μg/ml),TFL dose group(including 320 μg/ml),TFL low dose group(including 160 μg/ml)and blank control group.(1)MTT method is used to detect the absorbance(A value).Collected rat hepatic stellate cells were treated with different concentrations of TFL groups(160,320,640μg/ml TFL)and control group for 24 h,48h and 72 h in 96 well plate.There were five wells in each group.At the 490 nm wavelength,the A value was measured with enzyme standard meter.The inhibitory rate of rat hepatic stellate cells was calculated according to the absorbance value.(2)After acting for 72 h of TFL,the gene expression of MMP2 and TIMP1 in rat hepatic stellate cells was detected by PCR method.(3)The protein level of MMP2 and TIMP1 in rat hepatic stellate cells was detected by Western blotting method.Results: After exposure to TFL for 48,72 hours,produces obvious inhibition to rat hepatic stellate cells,increased with the extension of time and drug concentration and inhibition.There was significant difference(P<0.05)between TFL groups in 160,320,640μg/ml and control group.PCR indicated that TFL effect after 72 hours,MMP2 mRNA and TIMP1 mRNA in rat hepatic stellate cells has downgraded its role,and with increase of drug concentration,lower effect increased(P<0.05).Meanwhile,in rat hepatic stellate cells,expression of MMP2 and TIMP1 at the protein level was downregulated in the TFL treatment groups after exposure to different concentration TFL for 72 h.There was significant difference(P<0.05)between TFL treatment groups and control group.Conclusions: TFL can inhibit the proliferation of rat hepatic stellate cells and with the extension of time and drug concentration increase its inhibition enhanced gradually,the inhibition may be related to down-regulated expression of rat hepatic stellate cells at MMP2 and TIMP1 mRNA level and protein level.