荔枝核总黄酮对肝星状细胞增殖的抑制作用及其机制

摘要

Objective To explore the inhibitory effect of total flavonoids of lichi (TFL) on proliferation of rat hepatic stellate cells (HSC-T6) and to study the mechanism.Methods The cultured rat hepatic stellate cells in vitro were randomly divided into four groups as follows: the control group, low-dose, medium-dose, and high-dose TFL groups.Each group was cultured with cell culture medium (10%FBS included).TFL with final concentrations of 160, 320, and 640 μg/mL were added into the low-dose, medium-dose, and high-dose TFL groups.MTT method was used to detect the cell proliferation (A value) at 24, 48 and 72 h after treatment.After acting for 72 h, the mRNA and protein expression of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 1 (TIMP1) in rat hepatic stellate cells was detected by RT-PCR and Western blotting, respectively.Results As time went on, the A value of each group significantly decreased except the high-dose TFL group after exposure to TFL for 48 and 72 h (all P<0.05).After exposure for 24 h, the A value of HSC-T6 cells in each group had no obvious change;after exposure for 48 h, the A value of HSC-T6 cells in the low-dose, medium-dose, and high-dose TFL groups were lower than that of the control group, and the high-dose TFL group was the most significant (all P<0.05).After exposure for 72 h, the A value of HSC-T6 cells decreased with the increasing concentrations of TFL (all P<0.05).After 72 h, the mRNA and protein expression of MMP-2 and TIMP1 of the low-dose, medium-dose, and high-dose TFL groups was lower that that of the control group, and with the increase of drug concentration, the expression decreased gradually (all P<0.05).Conclusion TFL can inhibit the proliferation of HSC-T6 cells with a time-and dose-dependent manner, and the inhibition may be related to the down-regulation of MMP-2 and TIMP1 expression.%目的 探讨荔枝核总黄酮(TFL)对肝星状细胞(HSC)-T6增殖的抑制作用及其机制.方法 体外培养HSC-T6并随机分为对照组及TFL低、中、高剂量组,均置于含10% FBS的培养基中培养, TFL低、中、高剂量组均加入TFL,使终浓度分别为 160、320、640 μg/mL.分别于作用24、48、72 h时采用MTT法检测细胞增殖情况(吸光度值).作用72 h时采用RT-PCR法、Western blotting法检测基质金属蛋白酶2(MMP2)、组织金属蛋白酶抑制剂1(TIMP1)mRNA和蛋白表达.结果 除TFL高剂量组作用48、72 h外,各组随着作用时间的延长,HSC-T6吸光度值均明显增加(P均<0.05).作用24 h时,各组HSC-T6吸光度值变化不明显;作用48 h时,TFL低、中、高剂量组HSC-T6吸光度值明显低于对照组,以TFL高剂量组最显著(P均<0.05);作用72 h时,各TFL组随着TFL浓度增加HSC-T6吸光度值逐渐降低(P均<0.05).作用72 h时,TFL低、中、高剂量组MMP2、TIMP1 mRNA和蛋白相对表达量均低于对照组,且随着TFL浓度升高相对表达量逐渐降低(P均<0.05).结论 TFL可抑制HSC-T6增殖,具有时间和浓度依赖性;其作用机制可能与下调MMP2及TIMP1表达有关.