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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 effect of β-elemene on the expression and secretion of angiotensin II, angiotensinogen and Rho/ROCK in hepatic stellate cells. Methods: HSC-T6 cells were cultured for 24 h with different concentrations of β-elemene and blank control. After 4 hours, 12 hours and 24 hours, the amount of ANGII in the culture supernatant was detected by radioimmunoassay. RT-PCR was used to detect the expression of AGT and Rho/ROCK mRNA in HSC. RESULTS: At 4 h, the secretion of ANG II in the culture supernatant of 10.0 mg·L-1 β-elemene group was (50.970±8.081) pmol·L-1, which was lower than that of the blank group (74.500±10.999) pmol·L-1. Significantly decreased (P <0.05), and 5.0, 2.5mg · L-1β-elemene group had no significant inhibitory effect on ANGII secretion. At 12 h, the secretion of ANG II in 10,5 mg·L-1 β-elemene group was (83.727±6.850) and (91.090±3.226) pmol·L-1, respectively, which was significantly lower than that of the control group (104.367±5.030) pmol· L-1, (P<0.01, P<0.05); while 2.5mg·L-1 β-elemene had no significant effect. At 24 h, the secretion of ANG II in 5,2.5 mg·L-1β-elemene group was (104.133±3.296), (100.957±2.581) pmol·L-1, and that of blank group (116.620±7.110) pmol·L-1, respectively. Significantly decreased (P <0.05, P <0.01); and 10mg · L-1β-elemene group effect is not obvious. Compared with the blank group, at all time points of 4, 12, and 24 h, various concentrations of β-elemene had inhibitory effects on HSC-expressed AGT mRNA, with F being 30.33, 28.04, 107.19, and P being 0. 4, 12, 24 h respectively. At the time point, 2.5, 5.0, and 10 mg·L-1β-elemene inhibited the expression of RhoA mRNA in HSCs, and the values of F were 19.87, 14.35, and 40.13, respectively. All of them had no significant dose-dependence; the time point was 4, 24h. ,2.5,5.0,10 mg·L-1βElemene inhibited the expression of ROCK-1 and ROCK-2 mRNA in HSC (P<0.01), but there was no significant difference between the two groups at 12h. CONCLUSION: β-elemene can decrease the expression of AGT mRNA in ANGII and HSC secreted by HSCs, and inhibit the expression of downstream RhoA, ROCK-1, ROCK-2 mRNA, and inhibit the biological effects of ANGII, thus delaying the liver fibrosis. The occurrence of pulse pressure.