黄连碱对过氧化氢损伤的血管内皮细胞保护作用

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摘要目的探讨黄连碱对过氧化氢（H2O2）损伤的血管内皮细胞的保护作用及其机制。方法将体外培养的人脐静脉内皮细胞（EA.hy926）分为对照组、模型组、给药组。对照组加入新鲜培养基，模型组给H2O2刺激4 h，给药组用不同浓度的黄连碱（2.5、5、10、20、40 μmol/L）预处理4 h，再用H2O2刺激4 h。采用MTS法检测细胞活力，荧光探针-二氢乙啶（DHE）染色法检测细胞内活性氧（ROS），Annexin V-FITC/PI双染法检测细胞凋亡，Western blot检测凋亡相关蛋白caspase-3、Bcl-2和Bax的变化。结果与对照组比较，模型组细胞存活率和Bcl-2蛋白的表达明显降低，细胞内ROS含量、细胞凋亡率、caspase-3和Bax蛋白的表达明显升高（P < 0.01）；而与模型组比较，给药组细胞的存活率和Bcl-2蛋白的表达均随着给药浓度的增加而逐渐升高，细胞内ROS含量、细胞凋亡率、caspase-3和Bax蛋白的表达均随着给药浓度的增加而逐渐下降，差异均有高度统计学意义（P < 0.01）。结论黄连碱是通过抑制H2O2诱导的EA.hy926细胞存活率降低、细胞内ROS升高、细胞凋亡增多、caspase-3和Bax蛋白表达的上调以及Bcl-2蛋白表达的下调，发挥其对H2O2损伤的血管内皮细胞的保护作用，且呈剂量依赖性。

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	韦晟刘金春葛卫红

关键词 ：黄连碱, EA.hy926细胞, 氧化应激, 细胞凋亡

Abstract：Objective To investigate the protective effect and mechanism of coptisine on vascular endothelial cells induced by hydrogen peroxide (H2O2). Methods Human umbilical vein endothelial cells (EA.hy926) cultured in vitro were divided into control group, model group and drug administration group. The control group was added with fresh medium, and the model group was stimulated by H2O2 for 4 h. The drug-treated group was pretreated with different concentrations of coptisine (2.5, 5, 10, 20, 40 μmol/L) for 4 h, and then stimulated with H2O2 for 4 h. Cell viability was detected by MTS assay, intracellular reactive oxygen species (ROS) was detected by ROS fluorescent probe - dihydroethidine (DHE) staining, apoptosis was detected by Annexin V-FITC/PI double staining, and Western blot was used to detect the changes of apoptosis-related proteins caspase-3, Bcl-2 and Bax. Results Compared with the control group, the cell viability and Bcl-2 protein levels of the model group were markedly decreased, whereas the ROS levels, apoptotic rate, caspase-3 and Bax protein levels were significantly increased (P < 0.01). However, compared with the model group, the cell viability and Bcl-2 protein levels of dosing group were increased gradually with the increasing concentration of coptisine, while the ROS levels, apoptotic rate, caspase-3 and Bax protein levels were decreased gradually with the increasing concentration of coptisine (P < 0.01). Conclusion Coptisine exerts its protective effect on vascular endothelial cells injured by H2O2 in a dose-dependent manner by inhibiting the decrease of survival rate, the increase of ROS, the increase of apoptotic rate, the up-regulation of caspase-3 and Bax protein expression and the down-regulation of Bcl-2 protein expression in EA.hy926 cells induced by H2O2.

Key words： Coptisine EA.hy926 cell Oxidative stress Apoptosis

基金资助:江苏省医疗保险研究会科研课题（JSYB2017 002）。

通讯作者: 葛卫红（1962.12-），女，硕士，主任药师，主要从事临床药学研究。

作者简介: 韦晟（1980.10-），女，硕士，主要从事临床药理研究。

引用本文:

韦晟刘金春葛卫红. 黄连碱对过氧化氢损伤的血管内皮细胞保护作用[J]. 中国医药导报, 2019, 16(3): 4-7,21.
WEI Sheng LIU Jinchun GE Weihong. Protective effect of coptisine on vascular endothelial cells injured by hydrogen peroxide. 中国医药导报, 2019, 16(3): 4-7,21.

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