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目的 基于氧化应激及其相关凋亡蛋白探讨苓桂术甘汤对大鼠心肌缺血再灌注损伤的保护作用及机制。方法 45只SPF级SD雄性大鼠，适应性喂养1周，随机分成假手术组(Sham组)、心肌缺血再灌注组(IR组)、苓桂术甘汤预处理组[IR+LGZG(0.68 g/kg)组],每组15只。依据组别灌胃给予相应药物预处理，1次/d,连续7 d。第8天通过结扎大鼠心脏冠状动脉左前降支(ligating the anterior descending, LAD),心肌缺血30 min后，进行缺血再灌注24 h, Sham组只做穿线处理不结扎，建立MIRI大鼠模型。造模成功后分别取各组大鼠的腹主动脉血清和心脏组织进行相关指标的检测。通过心电图观察心功能变化，通过TTC染色观察心肌梗死面积，通过HE染色观察心肌组织病理改变。采用生化和ELISA法检测心肌酶损标志物Tn-T、CK-MB、LDH含量。采用ELISA法检测血清中氧化应激指标SOD、MDA、GSH-Px含量。采用Western blot法检测心肌组织中的凋亡蛋白Bax、Caspase-3、Bcl-2含量。结果 与Sham组相比，IR组大鼠的心电图ST段向上拱起明显，心肌组织病理形态学发生改变，心肌梗死面积显著增加(P<0.01),血清中Tn-T、CK-MB、LDH浓度显著升高(P<0.01),SOD表达显著降低(P<0.01),MDA表达显著升高(P<0.01),心肌细胞中Caspase-3、Bax蛋白表达上调(P<0.01),Bcl-2表达下调(P<0.01);与IR组相比，IR+LGZG组可以减少MIRI所致心脏损伤(P<0.05),升高SOD的表达(P>0.05),降低MDA的表达(P<0.01),减少MIRI引起的心肌细胞凋亡(P<0.05)。结论 苓桂术甘汤对大鼠心肌缺血再灌注损伤治疗作用佳，其作用机制部分是通过抑制氧化应激和随后的心肌细胞凋亡来实现的。
Abstract：Objective To explore the protective effect and mechanism of Linggui Zhugan Decoction(苓桂术甘汤,LGZG) on myocardial ischemia reperfusion injury(MIRI) in rats based on oxidative stress and its related apoptotic proteins. Methods Forty-five SPF male SD rats were randomly divided into sham operation group(Sham group),myocardial ischemia reperfusion group(IR group) and LGZG preconditioning group[IR+LGZG(0.68g/kg) group] with 15 rats in each group. The corresponding drug pretreatment was given by intragastric administration, once a day for 7 days. On the 8th day, the rat model of MIRI was established by ligating the left anterior descending branch of coronary artery(LAD). After 30 minutes of myocardial ischemia, the rats in the Sham group were subjected to IR for 24 hours. After successful modeling, the abdominal aorta serum and heart tissue of rats in each group were taken out to detect the related indexes. The changes of cardiac function were observed by electrocardiogram, the area of myocardial infarction was observed by TTC staining, and the pathological changes of myocardial tissue were observed by HE staining. Biochemical and ELISA methods were used to detect the contents of myocardial enzyme damage markers troponin T(Tn-T),creatine kinase isoenzyme(CK-MB) and lactate dehydrogenase(LDH). The contents of superoxide dismutase(SOD),malondialdehyde(MDA) and glutathione peroxidase(GSH-Px) in serum were detected by ELISA. The contents of apoptotic proteins Bax, caspase-3 and Bcl-2 in myocardial tissue were detected by Western blotting(WB). Results Compared with those in the Sham group, the ST segment of electrocardiogram(ECG) in the IR group arched upward obviously, the pathomorphology of myocardial tissue changed, the area of myocardial infarction increased significantly, the concentrations of Tn-T,CK-MB and LDH in serum increased significantly, the expression of SOD decreased significantly and the expression of MDA increased significantly in the IR group. The expressions of Caspase-3 and Bax protein were up-regulated and the expression of Bcl-2 was down-regulated in cardiomyocytes. Compared with the IR group, IR+LGZG group could reduce the cardiac injury induced by MIRI(P<0.05),increase the expression of SOD(P>0.05),decrease the expression of MDA(P<0.01) and reduce cardiomyocyte apoptosis induced by MIRI(P<0.05). Conclusions Linggui Zhugan Decoction has a better therapeutic effect on myocardial ischemia reperfusion injury in rats. The mechanism is partly through the inhibition of oxidative stress and subsequent cardiomyocyte apoptosis.
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基本信息:
DOI：10.13193/j.issn.1673-7717.2024.06.011
中图分类号:R285.5
引用信息:
[1]曾蕾,席晓荣,孙承霞,等.基于氧化应激及其相关凋亡蛋白研究苓桂术甘汤预处理对大鼠心肌缺血再灌注损伤的保护作用及机制[J].中华中医药学刊,2024,42(06):60-64+267-270.DOI:10.13193/j.issn.1673-7717.2024.06.011.
基金信息:
国家自然科学基金面上项目(82174355)
2024-06-10
2024-06-10