| 引用本文: |
何苗,李耀伟,王志琪,周志华,蒲科婷,丛梦静,王嘉星.基于细胞线粒体能量代谢研究甘草次酸拮抗乌头碱的心肌毒性作用[J].湖南中医药大学学报,2021,41(11):1650-1656[点击复制] |
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| 基于细胞线粒体能量代谢研究甘草次酸拮抗乌头碱的心肌毒性作用 |
| 何苗,李耀伟,王志琪,周志华,蒲科婷,丛梦静,王嘉星 |
| (湖南中医药大学研究生院, 湖南 长沙 410208;湖南省中药饮片标准化及功能工程技术研究中心, 湖南 长沙 410208;湖南省中药饮片标准化及功能工程技术研究中心, 湖南 长沙 410208;湖南中医药大学药学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 从线粒体能量代谢角度研究甘草次酸对乌头碱的心肌细胞毒性拮抗作用。方法 将大鼠心肌细胞H9c2细胞随机分为10组，即空白组、甘草次酸组、4个乌头碱组（乌头碱120、240、480、960 μmol/L）、4个乌头碱配伍甘草次酸组（不同浓度乌头碱分别与60 μmol/L甘草次酸配伍）。采用噻唑蓝（methyl thiazolyl tetrazolium，MTT）法测定不同浓度给药组对H9c2细胞的影响；采用ELISA法测定各组细胞三磷酸腺苷（adenosine 5’-triphosphate，ATP）含量、ATP酶活性水平；采用活性氧探针（2，7-dichlorodi-hydrofluorescein diacetate，DCFH-DA）检测细胞内活性氧（reactive oxygen species，ROS）、线粒体膜电位探针（5，5'6，6'-tetrachloro-1，1'，3，3'-tetraethyl-imidacarbocyanine iodide，JC-1）检测细胞内线粒体膜电位、钙离子荧光探针（rhod-2 acetoxymethyl ester，Rhod-2AM）检测细胞内钙离子浓度。结果 与单用乌头碱比较，乌头碱配伍甘草次酸后心肌细胞的ATP含量、Ca2+-ATP酶、Ca2+-Mg2+-ATP酶、Na+-K+-ATP酶活性升高，ROS含量减少，线粒体膜电位、线粒体内钙离子浓度降低（P<0.01或P<0.05）。结论 甘草次酸可拮抗乌头碱的心肌细胞毒性，其机制可能是与保护心肌细胞线粒体、增强线粒体能量代谢、维持钙稳态有关。 |
| 关键词: 线粒体能量代谢 乌头碱 甘草次酸 心肌毒性 钙稳态 |
| DOI：10.3969/j.issn.1674-070X.2021.11.002 |
| 投稿时间：2021-06-18 |
| 基金项目:国家自然科学基金项目（81503492）；湖南省自然科学基金项目（2015JJ6079，2021JJ80059）；湖南省教育厅重点项目（19A376）；长沙市自然科学基金项目（kq2014093）；国家级大学生创新项目（201910541019）。 |
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| Glycyrrhetinic Acid Against the Cardiotoxicity of Aconitine Based on Mitochondrial Energy Metabolism |
| HE Miao,LI Yaowei,WANG Zhiqi,ZHOU Zhihua,PU Keting,CONG Mengjing,WANG Jiaxing |
| (Graduate School, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Research Center of Standardization and Functional Engineering of Traditional Chinese Medicine in Hunan Province, Changsha, Hunan 410208, China;Research Center of Standardization and Functional Engineering of Traditional Chinese Medicine in Hunan Province, Changsha, Hunan 410208, China;School of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the antagonistic effect of glycyrrhetinic acid on cardiomyocytes cytotoxicity of aconitine from the perspective of mitochondrial energy metabolism. Methods The H9c2 cells of rat cardiomyocytes were randomly divided into 10 groups:blank group, glycyrrhetinic acid group, 4 aconitine groups (aconitine 120, 240, 480, 960 μmol/L), and 4 aconitine combined with glycyrrhetinic acid groups (aconitine in different concentrations was compatible with 60 μmol/L glycyrrhetinic acid). The effect of different concentrations on the H9c2 cell was determine by methyl thiazolyl tetrazolium (MTT). The content of adenosine5'-triphosphate (ATP) and the level of ATP enzyme activity in each group were measured by ELISA. The intracellular reactive oxygen species (ROS) was detected by 2,7-dichlorodi-hydrofluorescein diacetate (DCFH-DA), 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1) was used to detect intracellular mitochondrial membrane potential and rhod-2 acetoxymethyl ester (Rhod-2AM) probe was used to detect intracellular calcium ion concentration. Results Compared with aconitine alone, the ATP content, Ca2+-ATPase, Ca2+-Mg2+-ATPase and Na+-K+-ATPase activities of cardiomyocytes were significantly increased after aconitine combined with glycyrrhetinic acid. The ROS content, mitochondrial membrane potential and intracellular calcium concentration were significantly decreased (P<0.01 or P<0.05). Conclusion Glycyrrhetinic acid can reduce the cardiotoxicity of aconitine, which may be related to the protection of mitochondria of cardiomyocytes, the enhancement of mitochondrial energy metabolism and the maintenance of calcium homeostasis. |
| Key words: mitochondrial energy metabolism aconitine glycyrrhetinic acid cardiotoxicity calcium homeostasis |
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