β-榄香烯对血管紧张素Ⅱ诱导的HSC T6细胞增殖和迁移及RhoA的影响

目的 探讨β榄香烯对血管紧张素(ANG)Ⅱ诱导的肝星状细胞(HSC)增殖迁移及RhoA信号的影响. 方法体外培养HSC,应用ANGⅡ诱导HSC增殖迁移,采用四甲基偶氮唑盐法测定细胞增殖,Transwell小室检测细胞迁移,RT-PCR检测RhoA mRNA,Western blot检测RhoA蛋白.结果 1、2、4、8、10 μmol/L ANGⅡ组A490分别为0.129±0.004、0.143±0.016、0.166±0.012、0.183±0.004、0.283+0.014,与空白对照组(0.110+0.002)比较,ANGⅡ可显著促进HSC增殖,F=112.640,P<0.01;10、8、4 μmol/L的ANG Ⅱ可显著诱导HSC迁移,细胞迁移数分别为每高倍视野(17.40±2.07)、(12.60±1.14)、(9.00±1.58)个,与空白对照组[(1.60±0.55)个]比较,F=117.496,P<0.01.10、5、2.5mg/L的β榄香烯组A490分别为0.076±0.005、0.086±0.003、0.105±0.038,显著低于4μmol/L ANG Ⅱ组,F=95.706,P<0.01;10、5、2.5 mg/L的β-榄香烯组细胞迁移数分别为每高倍视野(1.33±0.58),(2.67±0.58)、(1.67±0.58)个,与4μmol/L ANGⅡ组比较,F=55.600,P<0.01,差异有统计学意义.4 μmol/L ANGⅡ组可显著诱导RhoA蛋白(相对表达量0.975±0.001)及mRNA(相对表达量0.951±0.045)表达,经10、5、2.5 mg/L的β-榄香烯作用后RhoA蛋白相对表达量分别为0.077±0.032、0.538 ± 0.026、0.701±0.078,RhoA mRNA相对表达量分别为0.734±0.038、0.845±0.036、0.881±0.027,较4 μ mol/L ANG Ⅱ组显著下降,F值分别为217.119,18.010,P值均<0.01.结论 ANG Ⅱ可显著诱导HSC增殖、辽移,随剂量的增加诱导作用加强,β-榄香烯可有效抑制ANG Ⅱ诱导的HSC增殖迁移,并能抑制HSC表达RhoA.

Effect of beta-elemene on the proliferation, migration and RhoA expression of hepatic stellate cells induced by angiotensin Ⅱ

Objective To explore the influence of beta-elemene on the proliferation, migration and RhoA expression of hepatic stellate cells (HSC) induced by angiotensin Ⅱ (ANG Ⅱ). Methods HSC were incubated in vitro. Proliferation and migration of the HSC were induced by ANG Ⅱ. The effect on the proliferation of HSC was determined by MTT colorimetry. The migration ability was detected by transwell chamber cultures. Total RNA was extracted by TRizol reagent and gene levels were determined by semi-quantitative RT-PCR. Protein levels were determined by Western blot. Results Different concentrations (from 1 to 10 μmol/L) orANG Ⅱ markedly promoted the growth of the HSC in a concentration dependent way (0 μ mol/L AN G Ⅱ, F = 112.640, P < 0.01). 10, 8, 4 μ mol/L ANGⅡ significantly induced HSC migration,F = 117.496, P < 0.01. Compared with the 4 μmol/L ANG Ⅱ group, 10 rag/L, 5 mg/L, 2.5 mg/L beta-elemene markedly inhibited HSC proliferation and migration induced by 4 μmol/L ANG Ⅱ (F values were 95.706 and 55.600 and P < 0.01). 4 μ mol/L ANG Ⅱ markedly promoted the protein and mRNA expressions of RhoA in HSC. 10 mg/L, 5 mg/L and 2.5 mg/L beta-elemene notably inhibited the expressions of RhoA protein and mRNA (F values were 217.119 and 18.010). Conclusion ANG Ⅱ can significantly induce the proliferation and migration of HSC. Beta-elemcne can inhibit the proliferation and migration of HSC induced by ANG Ⅱ.The effects of beta-elemone are mediated through inhibiting the RhoA signal transduction pathway and are associated with RhoA.