| 陈杭萍,董文珠,曹晓倩,等.合欢皮-白蒺藜药对对AngⅡ-ROS诱导下HSC-LX2细胞增殖及PKC/Nrf2/HO-1通路的影响[J].浙江中医药大学学报,2020,44(3):240-246, 251. |
| 合欢皮-白蒺藜药对对AngⅡ-ROS诱导下HSC-LX2细胞增殖及PKC/Nrf2/HO-1通路的影响 |
| Effect of Albizzia cortex-Tribulus terrestris Drug Pair on HSC-LX2 cell Proliferation and PKC/Nrf2/HO-1 Pathway Induced by AngⅡ-ROS |
| DOI：10.16466/j.issn1005-5509.2020.03.007 |
| 中文关键词: AngⅡ HSC-LX2细胞 肝纤维化 ROS 合欢皮 白蒺藜 |
| 英文关键词: AngⅡ HSC-LX2 cell liver fibrosis ROS Albizzia cortex Tribulus terrester |
| 基金项目: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) |
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| 中文摘要: |
| [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mRNA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。 |
| 英文摘要: |
| [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 AngⅡ, 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 low-dose group was inhibited significantly (all P＜0.05). Western blot results showed that compared with AngⅡgroup, 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 mRNA 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 AngⅡ 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. |
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