丹酚酸B对氧化应激损伤乳鼠心肌细胞的保护作用及其机制

doi:10.13481/j.1671-587x.20180515

摘要/Abstract

摘要： 目的：探讨丹酚酸B对氧化应激损伤乳鼠心肌细胞的保护作用，阐明其作用机制。方法：体外原代培养Wistar乳鼠心肌细胞，随机分为正常对照组、模型组及低和高剂量丹酚酸B组。除正常对照组外，其余各组培养基中加入终浓度为100 μmoL·L^-1的过氧化氢（H₂O₂）造成心肌细胞氧化损伤，其中低和高剂量丹酚酸B组分别加入终浓度为20和40 μmol·L^-1丹酚酸B。共同培养4 h后，光镜下观察心肌细胞形态表现，MTT法检测心肌细胞存活率；同时收集各组细胞及细胞培养液，分光光度法测定心肌细胞中还原型谷胱甘肽（GSH）水平、超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平，ELISA法测定细胞培养液中乳酸脱氢酶（LDH）及肌酸磷酸激酶（CK）水平。结果：与正常对照组比较，模型组部分心肌细胞悬浮在培养液中，贴壁细胞数量减少，伪足回缩，体积变小，心肌细胞自律搏动明显缓慢。与模型组比较，20和40 μmol·L^-1丹酚酸B组心肌细胞数量明显增多，伪足回缩少，自律搏动增强。与正常对照组比较，模型组心肌细胞存活率降低（P<0.01），心肌细胞中GSH水平降低（P<0.01），MAD水平升高（P<0.01），SOD活性降低（P<0.01），细胞培养液中LDH和CK水平升高（P<0.01）。与模型组比较，20和40 μmol·L^-1丹酚酸B组心肌细胞存活率升高（P<0.05或P<0.01），心肌细胞中GSH水平和SOD活性升高（P<0.05或P<0.01），MDA水平降低（P<0.05或P<0.01），细胞培养液中LDH和CK水平降低（P<0.05或P<0.01）。结论：丹酚酸B对心肌细胞氧化应激损伤有明显的保护作用，其作用机制可能与抑制过氧化反应和提高心肌细胞抗氧化能力有关。

关键词: 丹酚酸B, 心肌细胞, 氧化应激损伤, 心肌酶

Abstract: Objective:To investigate the protective effect of salvianolic acid B on the cardiomyocytes of the suckling rats with oxidative stress injury, and to clarify its mechanism. Methods:The cardiomyocytes of Wistar suckling ratswere primary cultured in vitro and randomly divided into normal control group, model group, low dose of salvianolic acid B group, and high dose of salvianolic acid B group. Except normal control group, the suckling rat cardiomyocytes were induced to oxidative stress injury by being exposed to H₂O₂ at the dose 100 μmoL·L^-1; 20 and 40 μmol·L^-1 salvianolic acid B were separatly added into low and high doses of salvianolic acid B groups. After co-culturing for 4 h, the morphological changes of cardiomyocytes were observed by light microscope;the survival rate of cardiomyocytes was examined using MTT. The levels of reductive glutathione (GSH), the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in the cardiomyocytes were detected with spectrophotometry. The levels of lactate dehydrogenase(LDH) and creatine kinase (CK) in culture medium were determined by ELISA. Results:Compared with normal control group, the cardiomyocytes in model group partially suspended in the culture medium, the number of adherent cells was decreased, the pseudopod contraction and the volume were decreased, and the self-discipline throb of cardiomyocytes was slow. Compared with model group,the number of cardiomyocytes in 20 and 40 μmol·L^-1 salvianolic acid B groups were increased significantly, the pseudopod contraction was reduced, and self-discipline throb was enhanced. Compared with normal control group, the survival rate of cardiomyocytes in model group was decreased(P<0.01), the level of GSH in cardiomyocytes was decreased(P<0.01), the level of MDA was increased (P<0.01),the activity of SOD was decreased(P<0.01), and the levels of CK and LDH in culture medium were increased (P<0.05 or P<0.01). Compared with model group, the survival rates of cardiomyocytes in 20 and 40 μmol·L^-1 salvianolic acid B groups were increased(P<0.05 or P<0.01), the levels of GSH and the activities of SOD were increased(P<0.05 or P<0.01), the levels of MDA were decreased (P<0.05 or P<0.01),and the CK and LDH levels in culture medium were decreased (P<0.05 or P<0.01). Conclusion:Salvianolic acid B can protect the cardiomyocytes against oxidative stress injury, and its mechanism may be associated with the inhibition of lipid peroxidatic reaction and the enhancement of antioxidant ability of cardiomyocytes.

Key words: salvianolic acid B, cardiomyocytes, oxidative stress injury, myocardial enzyme

中图分类号:

R283

陈雪, 沈楠, 安英, 徐博, 赵丽晶. 丹酚酸B对氧化应激损伤乳鼠心肌细胞的保护作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(05): 974-978.

CHEN Xue, SHEN Nan, AN Ying, XU Bo, ZHAO Lijing. Protective effect of salvianolic acid B on cardiomyocytes of suckling rats with oxidative stress injuryand its mechanism[J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 974-978.

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