摘要
[目的]探讨合欢皮-白蒺藜含药血清对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导产生的活性氧簇(reactive oxygen species,ROS)作用下人源性肝星状细胞HSC-LX2的增殖及蛋白激酶C(protein kinase C,PKC)/核因子E2相关因子2(nuclear factor erythroid-2 related factor 2,Nrf2)/血红素氧化酶-1(hemeoxygenase-1,HO-1)通路的影响。[方法]以HSC-LX2细胞为研究对象,先以AngⅡ诱导产生ROS,再以合欢皮-白蒺藜含药血清进行干预,MTT法检测HSC-LX2细胞的增殖情况,Western blot和Real-time PCR检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、PKC、Nrf2、HO-1蛋白以及m RNA表达情况。[结果]MTT结果显示,与AngⅡ组比较,合欢皮-白蒺藜药对高、中、低剂量组均显著抑制HSC-LX2细胞增殖(均P<0.05)。Western blot结果显示,与AngⅡ组比较,合欢皮-白蒺藜药对高、中、低剂量组α-SMA蛋白表达减少(均P<0.001),低剂量组PKC蛋白表达减少(P<0.05),高、中剂量组Nrf2蛋白表达减少(P<0.001,P<0.01),高、中剂量组HO-1蛋白表达减少(P<0.001,P<0.001)。Real-time PCR结果显示,与AngⅡ组比较,合欢皮-白蒺藜药对高、中、低剂量组α-SMA mRNA表达减少(P<0.01,P<0.001,P<0.001),PKC mRNA表达减少(均P<0.001),Nrf2 mRNA表达减少(P<0.001,P<0.001,P<0.05),高、中剂量组HO-1 mRNA表达减少(均P<0.001)。[结论]HSC-LX2细胞经AngⅡ刺激后增殖活化明显,且α-SMA表达升高,此过程与PKC、Nrf2、HO-1蛋白的激活有关。合欢皮-白蒺藜可通过抑制HSC-LX2细胞增殖来阻止肝纤维化,其机制与抑制PKC、Nrf2、HO-1的表达有关。
[Objective] To investigate the effect of Albizzia cortex and Tribulus terrester containing serum on protein kinase C(PKC)/nuclear factor erythroid-2 related factor 2(Nrf2)/hemeoxygenase-1(HO-1) pathway in human hepatic stellate cell HSC-LX2 under effect of reactive oxygen species(ROS) induced by angiotensin Ⅱ(AngⅡ). [Methods] HSC-LX2 cell was used as research object, ROS production was induced with Ang Ⅱ, and Albizzia cortex and Tribulus terrestris containing serum was used for intervention. The proliferation of HSC-LX2 cell was detected by MTT assay. The protein and mRNA expression of α-smooth muscle actin(α-SMA), PKC, Nrf2 and HO-1 was detected by Western blot and Real-time PCR. [Results]MTT results showed that compared with AngⅡ group, the proliferation of HSC-LX2 cell of Albizzia cortex and Tribulus terrestris high, medium and lowdose group was inhibited significantly(all P<0.05). Western blot results showed that compared with Ang Ⅱgroup, the portein expression of α-SMA of Albizzia cortex and Tribulus terrestris high, medium and low-dose group(all P<0.001),PKC of low-dose group(P<0.05), Nrf2 of high and medium-dose group(P<0.001,P<0.01), HO-1 of high and medium-dose group were inhibited(P<0.001,P<0.001).Real-time PCR results showed that compared with AngⅡgroup, the m RNA expression of α-SMA(P<0.01, P<0.001, P<0.001), PKC(all P<0.001), Nrf2(P<0.001, P<0.001, P<0.05) of Albizzia cortex and Tribulus terrestris high, medium and low-dose group, and HO-1 of high and medium-dose group were inhibited(all P <0.001). [Conclusion]The proliferation and activation of HSC-LX2 cells stimulated by Ang Ⅱ are obvious, and the expression of α-SMA increased, which was related to the activation of PKC, Nrf2 and HO-1 proteins. Albizzia cortex and Tribulus terrestris can inhibit liver fibrosis by inhibiting the proliferation of HSC-LX2 cell, and its mechanism is related to the inhibition of PKC, Nrf2 and HO-1 expression.
作者
陈杭萍
董文珠
曹晓倩
姚立
CHEN Hangping;DONG Wenzhu;CAO Xiaoqian(Zhejiang Chinese Medical University,Hangzhou(310053),China)
出处
《浙江中医药大学学报》
CAS
2020年第3期240-246,251,共8页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金项目(81173640)
浙江省自然科学基金项目(LY16H280009)
浙江中医药大学预研项目(2016ZG11)。
