黄芪注射液对睡眠剥夺大鼠心脏的保护作用及其机制

doi:10.13481/j.1671-587x.20200516

摘要/Abstract

摘要： 目的：观察黄芪注射液对睡眠剥夺大鼠血清心肌酶和氧化应激酶，心肌组织中炎症因子及心肌细胞凋亡相关蛋白表达的影响，探讨其心脏保护作用及机制。方法：将40只SD大鼠随机分为对照组、模型组、低和高剂量黄芪注射液组，每组10只。低和高剂量黄芪注射液组大鼠分别腹腔注射0.2和0.8 mL·kg^-1黄芪注射液，对照组和模型组大鼠给予生理盐水1 mL·100 g^-1，连续给药2周。采用改良多平台水环境法（MMPWM）建立睡眠剥夺大鼠模型；苏木精-伊红（HE）染色法观察各组大鼠心肌组织形态表现；原位末端标记法（TUNEL）检测大鼠凋亡心肌细胞，计算细胞凋亡指数（AI）；生化分析仪检测各组大鼠血清心肌酶中乳酸脱氢酶（LDH）、肌酸激酶（CK）、α-羟丁酸脱氢酶（α-HBDH）和丙氨酸氨基转移酶（ALT）活性；酶联免疫吸附测定法（ELISA）检测各组大鼠心肌组织中超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性及丙二醛（MDA）水平；ELISA法检测各组大鼠心肌组织中白细胞介素1β（IL-1β）、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）水平；Western blotting法检测各组大鼠心肌组织中凋亡因子B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、细胞间黏附分子1（ICAM-1）、血管细胞黏附分子1（VCAM-1）和含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase-3）蛋白表达水平。结果：与模型组比较，低和高剂量黄芪注射液组大鼠炎症浸润明显减轻，水肿程度减轻，AI明显降低（P<0.05），血清中LDH、CK、α-HBDH和ALT活性均明显降低（P<0.05），心肌组织中SOD和GSH-Px活性明显升高（P<0.05），MDA水平明显降低（P<0.05），心肌组织中IL-1β、IL-6和TNF-α水平均明显降低（P<0.05），心肌组织中Bax、ICAM-1、VCAM-1和Caspase-3蛋白表达水平明显降低（P<0.05），Bcl-2蛋白表达水平明显升高（P<0.05）；与低剂量黄芪注射液组比较，高剂量黄芪注射液组大鼠心肌组织损伤改善情况及各指标改变更加明显。结论：黄芪注射液可保护睡眠剥夺导致的大鼠心肌组织损伤，其机制可能与减轻氧化应激、下调黏附分子和凋亡蛋白表达，以及减少心肌细胞凋亡和炎症因子有关，在一定范围内呈剂量效应依赖性。

关键词: 黄芪注射液, 睡眠剥夺, 氧化损伤, 炎症因子, 细胞凋亡

Abstract: Objective: To observe the effects of Astragalus Injection on the serum myocardial enzymes,oxidative kinases, the expressions of inflammatory factors and the apoptosis-related proteins in myocardium tissue of the rats with sleep deprivation, and to investigate its protective effect on the heart and mechanism. Methods: A total of 40 SD rats were randomly divided into control group, model group, low dose of Astragalus Injection group and high dose of Astragalus Injection group, with 10 rats in each group. The rats in low and high doses of Astragalus Injection groups were intraperitioneally injected with 0.2 and 0.8 mL·kg^-1 Astragalus Injection, while the rats in control group and model group were given normal saline, and lasted for 2 weeks. The sleep deprivation rat models were established by modified multiple platform water environment method(MMPWM). The morphology of myocardum tissue of the rats in various groups was observed by HE staining. TUNEL method was used to detect the apoptotic myocardiocytes, and the apoptotic index (AI) was calculated. Biochemical analyzer was used to detect the activities of serum myocardial enzymes lactate dehydrogenase (LDH), creatine kinase (CK), α-hydroxybutyrate dehydrogenase (α-HBDH) and alanine aminotransferase (ALT). ELISA was used to detect the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the levels of malondialdehyde (MDA) in myocardium tissue of the rats in various groups. ELISA was used to detect the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in myocardium tissue of the rats in various groups. The expression levels of Bcl-2, Bax, intercellular cell adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1) and Caspase-3 proteins in myocardium tissue of the rats in various groups were determined by Western blotting method. Results: Compared with model group, the inflammatory infiltration and edema of the rats in low and high doses of Astragalus Injection groups were significantly reduced, the AI was significantly decreased (P<0.05), the activities of serum LDH, CK, α-HBDH and ALT of the rats in low and high doses of Astragalus Injection groups were significantly decreased (P<0.05), the activities of SOD and GSH-Px in myocardium tissue of the rats were significantly increased (P<0.05), the levels of MDA were significantly decreased (P<0.05), the levels of IL-1β, IL-6 and TNF-α in myocardium tissue were significantly decreased (P<0.05), the expression levels of Bax, ICAM-1, VCAM-1 and Caspase-3 proteins were significantly decreased (P<0.05), and the expression levels of Bcl-2 protein were significantly increased (P<0.05). Compared with low dose of Astragalus Injection group, the myocardial injury of the rats in high dose of Astragalus Injection group was improved significantly and the each index was changed more obviously. Conclusion: Astragalus Injection can protect the myocardium tissue damage caused by sleep deprivation of the rats, and its mechanism may be related to reducing oxidative stress, down-regulating the expressions of adhesion molecule and apoptotic protein and reducing apoptosis and inflammatory factor release; the effect is dose dependent in a certain range.

Key words: Astragalus Injection, sleep deprivation, oxidative damage, inflammatory factors, apoptosis

中图分类号:

R541

李伟, 张海峰, 王淼, 霍静, 张颖, 赵翠. 黄芪注射液对睡眠剥夺大鼠心脏的保护作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(05): 998-1003.

LI Wei, ZHANG Haifeng, WANG Miao, HUO Jing, ZHANG Ying, ZHAO Cui. Protective effect of Astragalus Injection on heart of rats with sleep deprivation and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 998-1003.

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