中国中药杂志

2021, v.46(06) 1467-1476

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人参-牡丹皮功效部位含药血清对H_2O_2诱导损伤的HUVEC的保护作用
Protective effect of serum containing ginseng and Moutan Cortex on HUVEC injured by H_2O_2

吴冰帆;张留记;李开言;王建霞;屠万倩;张钰;
WU Bing-fan;ZHANG Liu-ji;LI Kai-yan;WANG Jian-xia;TU Wan-qian;ZHANG Yu;Henan University of Chinese Medicine;Henan Academy of Traditional Chinese Medicine;Henan Luoyang Orthopedic-Traumatological Hospital (Henan Orthopedic Hospital);

摘要(Abstract):

探讨人参-牡丹皮功效部位含药血清对过氧化氢(H_2O_2)诱导损伤的人脐静脉内皮细胞(HUVEC)的影响。实验采用H_2O_2诱导损伤HUVEC,分为6组[空白组、模型组、人参总皂苷(TGG)组、丹皮总苷(TGM)组、丹皮酚(P)组、TGG+TGM+P组],各组药物与H_2O_2共培养24 h后,微量酶标法检测细胞内琥珀酸脱氢酶(SDH)、Ca~(2+)-Mg~(2+)-ATP酶的活性;流式细胞术检测各组细胞凋亡率及细胞内Ca~(2+)浓度、活性氧(ROS)及线粒体膜电位(JC-1)水平;Western blot法检测线粒体凋亡通路相关蛋白Bax, Bcl-2,cytochrome C,caspase-3,caspase-9的表达情况。结果显示H_2O_2对HUVEC有明显的损伤作用,使细胞SDH和Ca~(2+)-Mg~(2+)-ATP酶的活性显著降低(P<0.01),细胞凋亡+坏死率、JC-1下降率、ROS升高率和Ca~(2+)浓度升高率显著升高(P<0.01);人参-牡丹皮功效部位含药血清可不同程度地升高细胞内SDH和Ca~(2+)-Mg~(2+)-ATP酶活性(P<0.05或P<0.01),降低细胞凋亡+坏死率、JC-1下降率、ROS升高率和Ca~(2+)浓度升高率(P<0.05或P<0.01);同时下调线粒体凋亡通路相关蛋白Bax, caspase-3,caspase-9,cytochrome C表达水平,上调Bcl-2蛋白表达水平。结果表明,人参-牡丹皮功效部位含药血清对ROS所致血管内皮细胞损伤具有保护作用,其作用机制可能与改善线粒体功能,抑制线粒体凋亡通路被激活有关。
This study aimed to investigate the effect of serum containing ginseng and Moutan Cortex on human umbilical vein endothelial cells(HUVEC) injured with hydrogen peroxide(H_2O_2). HUVEC injured with H_2O_2 were divided into 6 groups, namely blank group, model group, ginsenoside(TGG) group, total glucosides of Moutan Cortex(TGM) group, paeonol(P) group and TGG+TGM+P group. After 24 hours of co-culture with H_2O_2, the activities of succinate dehydrogenase(SDH) and Ca~(2+)-Mg~(2+)-ATP were detected by microenzyme labeling. The apoptosis rate, intracellular Ca~(2+) concentration, reactive oxygen species(ROS) and mitochondrial membrane potential(JC-1) were detected by flow cytometry. The expressions of mitochondrial apoptosis pathway-related proteins Bax, Bcl-2, cytochrome C, caspase-3 and caspase-9 were detected by Western blot. The results showed that H_2O_2 could significantly damage HUVEC, decrease the activity of SDH and Ca~(2+)-Mg~(2+)-ATP(P<0.01), while could increase the apoptosis+necrosis rate, JC-1 decline rate, ROS increase rate and Ca~(2+) concentration increase rate(P<0.01). Serum containing ginseng and Moutan Cortex could increase the activities of SDH and Ca~(2+)-Mg~(2+)-ATP to different degrees, decrease the apoptosis+necrosis rate, JC-1 decline rate, ROS increase rate and Ca~(2+) concentration increase rate(P<0.05 or P<0.01), and down-regulate the protein expressions of Bax, caspase-3, caspase-9, cytochrome C, and up-regulate the protein expression of Bcl-2. The results showed that serum containing ginseng and Moutan Cortex has a protective effect on vascular endothelial cell injury induced by ROS, and its mechanism may be related to the improvement of mitochondrial function and the inhibition of the activation of mitochondrial apoptosis pathway.

关键词(KeyWords): 人参;牡丹皮;人脐静脉内皮细胞;线粒体
ginseng;Moutan Cortex;human umbilical vein endothelial cells;mitochondria

Abstract:

Keywords:

基金项目(Foundation): 河南省科技创新人才计划项目(144200510019);; 河南省中医药科学研究专项(2016ZY1003)

作者(Author): 吴冰帆;张留记;李开言;王建霞;屠万倩;张钰;
WU Bing-fan;ZHANG Liu-ji;LI Kai-yan;WANG Jian-xia;TU Wan-qian;ZHANG Yu;Henan University of Chinese Medicine;Henan Academy of Traditional Chinese Medicine;Henan Luoyang Orthopedic-Traumatological Hospital (Henan Orthopedic Hospital);

Email:

DOI: 10.19540/j.cnki.cjcmm.20201202.401

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