β-榄香烯抑制肝星状细胞表达ANGⅡ及RhoA/ROCK信号

β-elemene inhibits expression of ANGⅡ and RhoA/ROCK signaling in hepatic stellate Cells

目的:探讨β榄香烯对肝星状细胞表达分泌血管紧张素II、血管紧张素原及Rho/ROCK的影响.方法:HSC-T6细胞株培养24h,以不同浓度β榄香烯及空白对照分别作用4,12,24h后,放射免疫法检测培养上清中ANGⅡ的量,RT-PCR检测HSC中AGT及Rho/ROCK mRNA的表达。结果:在4h时,10.0mg·L-1β-榄香烯组培养上清中ANGII的分泌量为(50.970±8.081)pmol·L-1,较空白组(74.500±10.999)pmol·L-1明显下降(P<0.05),而5.0,2.5mg·L-1β-榄香烯组对ANGII分泌无明显抑制作用。12h时,10,5mg·L-1β-榄香烯组ANGⅡ分泌量分别为(83.727±6.850),(91.090±3.226)pmol·L-1,显著低于空白对照组(104.367±5.030)pmol·L-1,(P<0.01,P<0.05);而2.5mg·L-1β-榄香烯组作用不明显。24h时,5,2.5mg·L-1β榄香烯组ANGⅡ分泌量分别为(104.133±3.296),(100.957±2.581)pmol·L-1,与空白组(116.620±7.110)pmol·L-1比较明显减少(P<0.05,P<0.01);而10mg·L-1β-榄香烯组作用不明显。与空白组比较,4,12,24h各时间点,各个浓度的β榄香烯对HSC表达AGTmRNA均有抑制作用,F分别为30.33,28.04,107.19,P均为0。4,12,24h各时间点,2.5,5.0,10mg·L-1β榄香烯对HSC表达RhoA mRNA均有抑制作用,F分别为19.87,14.35,40.13,均P=0,无明显剂量依赖性;4,24h时间点,2.5,5.0,10mg·L-1β榄香烯对HSC表达ROCK-1,ROCK-2 mRNA均有抑制作用(P<0.01),而12h时各组之间无显著差异。结论:β榄香烯能使HSC分泌ANGⅡ及HSC内AGT mRNA表达降低,同时抑制下游信号RhoA,ROCK-1,ROCK-2 mRNA的表达,抑制ANGⅡ的生物学效应,从而延缓肝纤维化及门脉高压的发生。

Objective:To investigate the influence of β-Elemene on expression of ANGⅡ and RhoA/ROCK signaling in Hepatic Stellate Cells.Method:In vitro,HSC-T6 cell line was cultured for 24 hours and treated with several concentration of β-elemen(e5.0,5.0,2.5 mg·L-1) and Y-27632(30 μmol·L-1) for 4,12 and 24 h.Secretion of ANGⅡ in the supernatant was detected by Radioimmunoassay.The mRNA expression of AGT,RhoA,ROCK-1 and ROCK-2 for 4 h,12 h,24 h was detected by RT-PCR respectively.Result:On the time point of 4h,the secre...