芪参益气滴丸对慢性心力衰竭大鼠心肌细胞凋亡的抑制作用及其机制

doi:10.13481/j.1671-587x.20200512

摘要/Abstract

摘要： 目的：探讨芪参益气滴丸（QSYQ）对慢性心力衰竭（CHF）大鼠心肌细胞凋亡的影响，并阐明其可能的作用机制。方法：60只雄性SD大鼠随机分为假手术组、模型组、阳性药对照组（6.75 mg·kg^-1·d^-1卡托普利）、低剂量（135 mg·kg^-1·d^-1）QSYQ组和高剂量（270 mg·kg^-1·d^-1）QSYQ组，每组12只。采用冠状动脉前降支结扎法建立CHF模型，造模成功后连续灌胃给药4周，超声心动图检测大鼠心功能，采用脱氧核糖核苷酸末端转移酶介导的缺口末端标志法（TUNEL）检测大鼠心肌细胞凋亡率，比色法测定大鼠血清中乳酸脱氢酶（LDH）和超氧化物歧化酶（SOD）活性及丙二醛（MDA）水平，流式细胞术检测大鼠心肌组织中活性氧（ROS）水平，Western blotting法检测大鼠心肌组织中凋亡相关蛋白和核因子E2相关因子2（Nrf2）及血红素氧化酶1（HO-1）蛋白表达水平。结果：与假手术组比较，模型组大鼠左室收缩末期内径（LVSD）和左室舒张末期内径（LVDD）明显增加（P<0.05），左室射血分数（EF）和左室短轴缩短率（FS）明显降低（P<0.05），棕黄色心肌细胞数明显增多（P<0.05），细胞凋亡率明显升高（P<0.05），血清中LDH活性及ROS和MDA水平明显升高（P<0.05），SOD活性明显降低（P<0.05），心肌组织中活化半胱氨酸天冬氨酸蛋白酶3（Cleaved-Caspase-3）和B淋巴细胞瘤2（Bcl-2）相关X蛋白（Bax）表达水平明显升高（P<0.05），Bcl-2、Nrf2和HO-1蛋白水平明显降低（P<0.05）；与模型组比较，高剂量QSYQ组和阳性药对照组大鼠LVSD和LVDD明显降低（P<0.05），EF和FS明显升高（P<0.05），棕黄色心肌细胞数明显减少（P<0.05），细胞凋亡率明显降低（P<0.05），血清中LDH活性及ROS和MDA水平明显降低（P<0.05），SOD活性明显升高（P<0.05），心肌组织中Cleaved-Caspase-3和Bax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），而低剂量QSYQ组大鼠上述各相关指标差异无统计学意义（P>0.05）。与模型组比较，高剂量QSYQ组大鼠心肌组织中Nrf2和HO-1蛋白表达水平明显升高（P<0.05），阳性药对照组和低剂量QSYQ组大鼠心肌组织中Nrf2和HO-1蛋白表达水平差异无统计学意义（P>0.05）。结论：QSYQ可抑制CHF大鼠心肌细胞凋亡，其作用机制可能与激活Nrf2/HO-1信号通路、降低氧化损伤有关。

关键词: 慢性心力衰竭, 芪参益气滴丸, 心肌细胞, 核因子E2相关因子2/血红素氧化酶-1信号通路

Abstract: Objective: To investigate the effect of Qishen-Yiqi Dropping Pill (QSYQ) on the apoptosis of myocardial cells in the rats with chronic heart failure (CHF), and to explore its possible mechanism. Methods: Sixty male SD rats were randomly divided into sham operation group, model group, positive drug control group (6.75 mg·kg^-1·d^-1 Captopril), low dose (135 mg·kg^-1·d^-1) of QSYQ group and high dose (270 mg·kg^-1·d^-1) of QSYQ group, and there were 12 rats in each group. The CHF models were established by ligating anterior descending coronary artery. After successful establishment of the models,the rats were continuously administrated by gavage for 4 weeks; the cardiac function was measured by echocardiography and the apoptotic rate of myocardial cells was detected by TUNEL assay. The activities of serum lactate dehydrogenase (LDH) and superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) of the rats were determined by colorimetry, the reactive oxygen species (ROS) levels in myocardium tissue of the rats were detected by flow cytometry, and the expression levels of apoptosis-related proteins, Nrf2 and HO-1 proteins in myocardium tissue of the rats were detected by Western blotting method. Result: Compared with sham operation group, the left ventricular end-systolic diameter (LVSD) and left ventricular end-diastolic diameter (LVDD) of the rats in model group were significantly increased (P<0.05), and the left ventricular ejection fraction (EF), and left ventricular short-axis shortening rate (FS) were significantly decreased (P<0.05); the number of brown and yellow myocardial cells was increased(P<0.05), and the apoptotic rate was significantly increased (P<0.05);the LDH activity and the ROS and MDA levels in serum were significantly increased (P<0.05),and the SOD activity was significantly decreased (P<0.05); the expression levels of Cleaved-Caspase-3 and Bax protein in myocardium tissue were significantly increased (P<0.05), and the expression levels of Bcl-2, Nrf2 and HO-1 proteins were significantly decreased (P<0.05). Compared with model group, the LVSD and LVDD of the rats in high dose of QSYQ group and positive drug control group were significantly decreased (P<0.05), EF and FS were significantly increased (P<0.05); the number of brown and yellow myocardial cells was decreased (P<0.05), the apoptotic rates were significantly decreased (P<0.05), the LDH activities and the ROS and MDA levels in serum were significantly decreased (P<0.05), and the SOD activities were significantly increased (P<0.05); the expression levels of Cleaved-Caspase-3 and Bax proteins in myocardium tissue of the rats were significantly decreased (P<0.05), and the expression levels of Bcl-2 protein were significantly increased (P<0.05); there were no significant differences in the above indexes of the rats between low dose of QSYQ group and model group (P>0.05). Compared with model group, the expression levels of Nrf2 and HO-1 proteins in myocardium tissue of the rats in high dose of QSYQ group were significantly increased (P<0.05); there were no significant differences in the expression levels of Nrf2 and HO-1 proteins in myocardium tissue of the rats between low dose of QSYQ group and positive drug group (P>0.05). Conclusion: QSYQ can inhibit the apoptosis of myocardial cells in the rats with CHF, and its mechanism may be related to activating the Nrf2/HO-1 signaling pathway and reducing the oxidative damage.

Key words: chronic heart failure, Qishen-Yiqi Dropping Pill, myocardial cells, nuclear factor E2/Heme oxidase-1 signaling pathway

中图分类号:

R541.6

陈家显, 刘先霞, 陈跃武, 陈磊, 张远生, 陈劲松. 芪参益气滴丸对慢性心力衰竭大鼠心肌细胞凋亡的抑制作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(05): 972-978.

CHEN Jiaxian, LIU Xianxia, CHEN Yuewu, CHEN Lei, ZHANG Yuansheng, CHEN Jinsong. Inhibitory effect of Qisheng-Yiqi Dropping Pill on apoptosis of myocardial cells in rats with chronic heart failure and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 972-978.

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