摘要
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、640μg/m L。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。
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.
出处
《山东医药》
CAS
北大核心
2017年第24期9-12,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81360530)
关键词
肝纤维化
荔枝核总黄酮
肝星状细胞
基质金属蛋白酶2
组织金属蛋白酶抑制剂1
hepatic fibrosis
total flavonoids of lichi
hepatic stellate cells
matrix metalloproteinase 2
tissue inhibi-tor of metalloproteinase 1
