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

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

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，RTPCR检测RhoAmRNA，Westernblot检测RhoA蛋白。结果1、2、4、8、10μmol／LANGⅡ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、4μmol／L的ANG11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D榄香烯组4490分别为0．076±0．005、0．086±0．003、0．105±0．038，显著低丁4mol／LANGⅡ组，F=95．706，P〈0．01；10、5、2．5mg／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／LANGⅡ组比较，F=55．600，P〈0．01，差异有统计学意义。4μmol／LANGⅡ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、2．5mg／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4μmol／LANGⅡ组始著下降，F值分别为217．119，18．010，P值均〈0．01。结论ANGⅡ可显著诱导HSC增殖、迁移，随剂量的增加诱导作用加强，β-榄香烯可有效抑制ANGⅡ诱导的HSC增殖迁移，并能抑制HSC表达RhoA。

Objective To explore the influence ofbeta-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 10μmol/L） ofANG Ⅱ markedly promoted the growth of the HSC in a concentration dependent way （0 μmol/L ANG Ⅱ, F = 112.640, P 〈 0.01）. 10, 8, 4 μmol/L ANGⅡ significantly induced HSC migration, F = 117.496, P 〈 0.01. Compared with the 4 μmol/L ANG II group, 10 mg/L, 5 nag/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）. 4μ 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 ofHSC. Beta-elemene can inhibit the proliferation and migration of HSC induced by ANG Ⅱ. The effects of beta-elemene are mediated through inhibiting the RhoA signal transduction pathway and are associated with RhoA.