作者：尹梦影，曹伟，胡鑫，郑景辉，莫云秋

单位：

1.广西中医药大学2.广西中医药大学附属瑞康医院老年病科

Units：

1.Guangxi University of Chinese Medicine, Nanning 530001, China 2.Department of Gastroenterology, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530011, China

关键词：

心肌再灌注损伤; 心肌缺血再灌注损伤; 大鼠; 养心通脉有效部位方; 线粒体;

Keywords：

Myocardial reperfusion injury; Myocardial ischemic reperfusion injury; Rats; Active principle region of Yangxin Tongmai formula; Mitochondria

CLC：

DOI：

10.12114/j.issn.1008-5971.2023.00.195

Funds：

国家自然科学基金资助项目（82160875）

摘要：

目的 分析不同剂量养心通脉有效部位方（apr-YTF）对心肌缺血再灌注损伤（MIRI）模型大鼠心肌线粒体代谢相关酶及其转运的影响。方法 本实验时间为2020年6月至2021年6月。将70只健康雄性Wistar大鼠随机分为空白对照组、模型组、阴性对照组、阳性对照组及apr-YTF低、中、高剂量组，每组10只。除空白对照组外，其他组大鼠构建MIRI模型，造模期间，各组大鼠共死亡25只，为保证实验完整性，补充造模至每组10只。造模完成后，空白对照组及模型组大鼠正常饲养，阴性对照组大鼠给予0.9%氯化钠溶液灌胃，阳性对照组大鼠给予冠心苏合丸灌胃，apr-YTF低、中、高剂量组大鼠分别给予1.5、3.0、6.0 ml apr-YTF灌胃，1次/d，共干预2周。比较空白对照组和模型组造模24 h后血液流变学指标（包括全血低切、中切、高切黏度）、血清心肌梗死标志物[肌酸激酶同工酶（CKMB）、心肌肌钙蛋白I(cTnI)、乳酸脱氢酶（LDH）]水平。比较七组大鼠造模第14天心肌组织病理学检查结果、线粒体代谢相关酶[ATP、磷酸果糖激酶1(PFK1)、琥珀酸脱氢酶（SDH）、腺苷酸转运体（ANT）、异枸橼酸脱氢酶（IDH）]水平及心肌组织中肿瘤坏死因子α诱导蛋白2(TNFαIP2)、MIRO1表达水平。结果 模型组全血低切、中切黏度及血清CK-MB、cTnI、LDH水平高于空白对照组（P<0.05）。模型组ATP水平低于空白对照组，模型组、阴性对照组PFK1、SDH、ANT、IDH水平高于空白对照组（P<0.05）；阴性对照组、阳性对照组及apr-YTF低、中、高剂量组ATP水平高于模型组，阳性对照组及apr-YTF中、高剂量组PFK1、ANT、IDH水平低于模型组，阳性对照组及apr-YTF低、中、高剂量组SDH水平低于模型组（P<0.05）;apr-YTF低剂量组ATP、ANT水平高于阳性对照组，apr-YTF低、中剂量组PFK1、IDH水平高于阳性对照组（P<0.05）;apr-YTF中剂量组ATP水平低于apr-YTF低剂量组，apr-YTF中、高剂量组PFK1、ANT、IDH水平低于apr-YTF低剂量组，apr-YTF高剂量组SDH水平低于apr-YTF低剂量组（P<0.05）。模型组、阴性对照组心肌组织中TNFαIP2表达水平高于空白对照组，心肌组织中MIRO1表达水平低于空白对照组（P<0.05）；阳性对照组及apr-YTF中、高剂量组心肌组织中TNFαIP2表达水平低于模型组，阳性对照组及apr-YTF低、中、高剂量组心肌组织中MIRO1表达水平高于模型组（P<0.05）;apr-YTF低剂量组心肌组织中TNFαIP2表达水平高于阳性对照组，apr-YTF低、中、高剂量组心肌组织中MIRO1表达水平低于阳性对照组（P<0.05）;apr-YTF中、高剂量组心肌组织中TNFαIP2表达水平低于apr-YTF低剂量组（P<0.05）。结论 不同剂量apr-YTF均可有效改善MIRI模型大鼠心肌线粒体代谢相关酶，进而减轻线粒体能量代谢功能障碍，其还可减轻线粒体转运障碍；且中剂量（3.0 ml）、高剂量（6.0 ml）apr-YTF的效果相似，均优于低剂量（1.5 ml）apr-YTF。

Abstract：

Objective To analyze the effects of different doses of active principle region of Yangxin Tongmai formula (apr-YTF) on myocardial mitochondrial metabolism-related enzymes and their transport in rats with myocardial ischemia?reperfusion injury (MIRI) . Methods This experiment was conducted from June 2020 to June 2021. Seventy healthy male Wistar rats were randomly divided into blank control group, model group, negative control group, positive control group, apr?YTF low dose group, apr-YTF medium dose group and apr-YTF high dose group, with 10 rats in each group. Except for the blank control group, the MIRI model was constructed in the other groups of rats. During the modeling period, a total of 25 rats in each group died. To ensure the integrity of the experiment, membrane building was supplemented to 10 rats in each group. After the completion of modeling, the rats in the blank control group and model group were fed normally, the rats in the negative control group were given 0.9% sodium chloride solution by gavage, the rats in the positive control group were given Guanxin Suhe pill by gavage, and the rats in the apr-YTF low-dose group, apr-YTF medium-dose group and apr-YTF high-dose group were given 1.5, 3.0 and 6.0 ml apr-YTF by gavage, once a day, for a total of 2 weeks of intervention. The hemorheological indexes (including whole blood low-cut, mid-cut and high-cut viscosity) and serum myocardial infarction markers [creatine kinase isoenzyme (CK?MB) , cardiac troponin I (cTnI) and lactate dehydrogenase (LDH) ] levels were compared between the blank control group and the model group 24 h after modeling. The myocardial histological examination results, the levels of mitochondrial metabolism?related enzymes [ATP, phosphofructokinase 1 (PFK1) , succinate dehydrogenase (SDH) , adenine nucleotide translocator (ANT) and isocitrate dehydrogenase (IDH) ] and the expression levels of tumor necrosis factor alpha-induced protein 2 (TNFαIP2) and MIRO1 in the myocardium of seven groups were compared on the 14th day after modeling. Results The whole blood low-cut and mid-cut viscosity and serum CK-MB, cTnI and LDH levels in the model group were higher than those in the blank control group (P < 0.05) . The ATP levels in the model group were lower than those in the blank control group, and the PFK1, SDH, ANT, and IDH levels in the model and negative control groups were higher than those in the blank control group (P < 0.05) ; the ATP levels in the negative control group, positive control group, apr-YTF low-dose group, apr-YTF medium-dose group, and apr-YTF high-dose group were higher than those in the model group, the PFK1, ANT, and IDH levels in the positive control group, apr?YTF medium-dose group, and apr-YTF high-dose group were lower than those in the model group, the SDH levels in the positive control group, apr-YTF low-dose group, apr-YTF medium-dose group, and apr-YTF high-dose group were lower than those in the model group (P < 0.05) ; ATP and ANT levels in apr-YTF low-dose group were higher than those in positive control group, and the PFK1 and IDH levels in apr-YTF low-dose group and apr-YTF medium-dose group were higher than those in positive control group (P < 0.05) ; the ATP levels in the apr-YTF medium-dose group were lower than those in the apr-YTF low-dose group, while the PFK1, ANT, and IDH levels in the apr-YTF medium-group and apr-YTF high-dose group were lower than those in the apr-YTF low-dose group, the SDH levels in the apr-YTF high-dose group were lower than those in the apr-YTF low-dose group (P < 0.05) . The expression level of TNF-αIP2 in myocardial tissue of model group and negative control group was higher than that of blank control group, and the expression level of MIRO1 in myocardial tissue was lower than that of blank control group (P < 0.05) ; the expression level of TNF-αIP2 in myocardial tissue of positive control group and apr-YTF medium-dose group and apr-YTF high-dose group was lower than that of model group, and the expression level of MIRO1 in myocardial tissue of positive control group and apr-YTF low-dose group, apr-YTF medium-dose group and apr-YTF high-dose group was higher than that of model group (P < 0.05) ; the expression level of TNF-αIP2 in myocardial tissue of apr-YTF low-dose group was higher than that of positive control group, and the expression level of MIRO1 in myocardial tissue of apr-YTF low-dose group, apr-YTF medium?dose group and apr-YTF high-dose group was lower than that of positive control group (P < 0.05) ; the expression level of TNF?αIP2 in myocardium of apr-YTF medium-dose group and apr-YTF high-dose group was lower than that of apr-YTF low?dose group (P < 0.05) . Conclusion Different doses of apr-YTF can effectively improve myocardial mitochondrial metabolism? related enzymes in MIRI model rats, thereby alleviating mitochondrial energy metabolism dysfunction and mitochondrial transport dysfunction. And the effects of medium dose (3.0 ml) and high dose (6.0 ml) apr-YTF were similar, and both were better than that of low dose (1.5 ml) apr-YTF.

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