Yixin Formula modulates SIRT1/PGC⁃1α signaling pathway to improve myocardial ischemia⁃reperfusion injury in rats

DONG Li; YANG Yanling; ZHENG Yufan; RONG Jingfeng

doi:10.16305/j.1007-1334.2023.2210071

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Yixin Formula modulates SIRT1/PGC⁃1α signaling pathway to improve myocardial ischemia⁃reperfusion injury in rats

更新时间：2023-07-15

- Yixin Formula modulates SIRT1/PGC⁃1α signaling pathway to improve myocardial ischemia⁃reperfusion injury in rats
- Shanghai Journal of Traditional Chinese Medicine Vol. 57, Issue 7, Pages: 58-65(2023)
- 作者机构：
  
  1.上海中医药大学附属曙光医院心血管内科（上海 201203）
  2.上海中医药大学附属曙光医院心血管病研究室（上海 201203）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2023.2210071
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董莉,杨燕玲,郑钰凡,等.益心方调控SIRT1/PGC⁃1α信号通路改善大鼠心肌缺血再灌注损伤[J].上海中医药杂志,2023,57(7):58-65.

DONG Li,YANG Yanling,ZHENG Yufan,et al.Yixin Formula modulates SIRT1/PGC⁃1α signaling pathway to improve myocardial ischemia⁃reperfusion injury in rats[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(7):58-65.
董莉,杨燕玲,郑钰凡,等.益心方调控SIRT1/PGC⁃1α信号通路改善大鼠心肌缺血再灌注损伤[J].上海中医药杂志,2023,57(7):58-65. DOI： 10.16305/j.1007-1334.2023.2210071.

DONG Li,YANG Yanling,ZHENG Yufan,et al.Yixin Formula modulates SIRT1/PGC⁃1α signaling pathway to improve myocardial ischemia⁃reperfusion injury in rats[J].Shanghai Journal of Traditional Chinese Medicine,2023,57(7):58-65. DOI： 10.16305/j.1007-1334.2023.2210071.

摘要

目的,2,探讨中药复方益心方改善心肌缺血再灌注损伤大鼠的作用及其机制。,方法,2,将60只清洁级Wistar大鼠随机分为4组，即假手术组、心肌缺血再灌注组（模型组）、益心方预处理+心肌缺血再灌注组（益心方组）、益心方预处理+心肌缺血再灌注+ 沉默信息调节因子1（SIRT1）抑制剂组（益心方+抑制剂组），每组15只。各组大鼠适应性喂养7 d后，益心方组和益心方+抑制剂组给予益心方（10 mL/kg）灌胃7 d，其余各组灌胃等体积的质量分数为0.9%的氯化钠溶液。术前3 d及术前20 min，益心方+抑制剂组尾静脉注射SIRT1抑制剂sirtinol（2 mg/kg），其余各组尾静脉注射等体积的质量分数为0.9%的氯化钠溶液。假手术组只穿线不结扎；其余各组结扎冠状动脉左前降支30 min，建立心肌缺血损伤模型，术中观察心电图变化情况。再灌注后检测心电图，观察再灌注模型是否建立成功；采用超声心动图和血流动力学检测评价各组大鼠的心功能；苏木精-伊红（HE）染色法观察心肌组织病理形态学的变化；2,3,5-氯化三苯基四氮唑（TTC）染色法测定心肌梗死面积；原位末端转移酶标记技术（TUNEL）法检测心肌细胞凋亡指数；生化法检测各组大鼠血清中的肌酸激酶（CK）、乳酸脱氢酶（LDH）水平；酶联免疫吸附试验（ELISA）检测各组大鼠血清超氧化物歧化酶（SOD）、丙二醛（MDA）、过氧化氢（H,2,O,2,）活性；免疫组织化学法和Western blot法检测SIRT1、过氧化物酶体增殖活化受体γ辅助活化因子-1α（PGC-lα）、B淋巴细胞瘤-2蛋白（Bcl-2）、B淋巴细胞瘤-2相关X蛋白（Bax）表达情况。,结果,2,①与假手术组比较，模型组大鼠心电图ST段明显抬高，表明大鼠心肌缺血模型建立成功；超声心动图和血流动力学检测显示，模型组大鼠左心室射血分数（LVEF）、左心室短轴缩短率（LVFS）、左心室收缩压（LVSP）、左心室内压最大上升速率（+dp/dtmax）降低，左心室舒张末压（LVDP）、左心室内压最大下降速率（-dp/dtmax）升高（,P,<,0.05）；TTC染色结果显示，模型组大鼠心肌梗死面积增加（,P,<,0.05）；生化检测结果显示，模型组大鼠血清中CK、LDH、MDA、H,2,O,2,水平升高，SOD水平降低（,P,<,0.05）；免疫组织化学法和Western blot检测结果显示，模型组大鼠心肌组织中Bcl-2、SIRT1、PGC-1α表达下降，Bax表达升高（,P,<,0.05）。②与模型组比较，益心方组心肌梗死的面积减少，LVEF、LVFS、LVSP、+dp/dtmax升高，LVDP、-dp/dtmax降低，左心室射血量增加，心功能改善（,P,<,0.05）；血清CK、LDH、MDA、H,2,O,2,水平降低， SOD水平升高（,P,<,0.05）；心肌组织中Bcl-2、SIRT1、PGC-1α表达升高，Bax表达降低（,P,<,0.05）。③与益心方组比较，益心方+抑制剂组大鼠血清CK、LDH、MDA、H,2,O,2,水平升高，SOD水平降低（,P,<,0.05）；心肌组织中Bcl-2、SIRT1、PGC-1α表达降低、Bax表达升高（,P,<,0.05）。,结论,2,益心方预处理能够减轻氧化应激损伤和细胞凋亡，改善心肌缺血再灌注损伤，其作用机制可能与调控SIRT1/PGC-1α信号通路相关。

Abstract

Objective,2,To observe the effect of Traditional Chinese medicine compound Yixin Formula（YXF） to improve myocardial ischemia-reperfusion injury（MIRI）in rats and its mechanism.,Methods,2,Sixty clean-grade Wistar rats were randomly divided into four groups： sham-operated group， myocardial ischemia-reperfusion group （model group）， Yixin Formula+myocardial ischemia-reperfusion group （YXF group）， Yixin Formula+myocardial ischemia-reperfusion+SIRT1 inhibitor group （YXF+inhibitor group）， 15 rats in each group. After adaptive feeding of rats in each group for 7 d， YXF and YXF + inhibitor groups were given YXF 10 mL/kg by gavage for 7 d， and the remaining groups were given an equal volume of 0.9% sodium chloride solution by gavage. Three days and 20 min before surgery， an inhibitor of SIRT1， sirtinol（2 mg/kg）， was injected into the tail vein of rats in the YXF + inhibitor group， and an equal volume of 0.9% sodium chloride solution was injected into the tail vein of rats in the other groups. The sham-operated group was only threaded without ligation， while the left anterior descending branch of the coronary artery was ligated for 30 min in the remaining groups to establish a model of myocardial ischemic injury， and the changes of ECG were observed during operation. After reperfusion， ECG was examined to observe whether the reperfusion model was successfully established， and echocardiography and hemodynamics were used to examine the cardiac function of rats in each group. HE staining was used to observe the histomorphological changes of myocardial tissue. TTC staining was used to determine the area of myocardial infarction. TUNEL method was used to detect the apoptosis rate of myocardial cells. The biochemical method was used to detect the levels of CK and LDH in the serum of rats in each group. ELISA was used to detect the levels of SOD， MDA and H,2,O,2 ,in the serum of rats in each group. The protein expression of SIRT1， PGC-lα， Bcl-2 and Bax was detected by immunohistochemistry and Western blot.,Results,2,① Compared with the sham-operated group， the ST segment of the ECG was significantly elevated in the model group， indicating that the rat myocardial ischemia model was successfully established. Echocardiography and hemodynamic tests showed that LVEF， LVFS， LVSP， +dp/dtmax were decreased and LVDP， -dp/dtmax were increased in the model group （,P,<,0.05）. TTC staining showed that the myocardial infarct area was increased in the model group （,P,<,0.05）. The biochemical results showed that the serum levels of CK， LDH， MDA， and H,2,O,2, were increased， and the level of SOD was decreased in the model rats （,P,<,0.05）. The immunohistochemical and Western blot results showed that the expressions of Bcl-2， SIRT1 and PGC-1α proteins were decreased， and Bax expression was increased in the myocardial tissue of the model rats （,P,<,0.05）. ② Compared with the model group， the YXF could reduce the area of myocardial infarction. The levels of LVEF， LVFS， LVSP， +dp/dtmax were increased， the levels of LVDP， -dp/dtmax were decreased， the left ventricular ejection volume was increased， and the cardiac function was improved in the YXF group （,P,<,0.05）. The levels of CK， LDH， MDA， H,2,O,2, were decreased and the level of SOD was increased in the YXF group. The expressions of Bcl-2， SIRT1 and PGC-1α in myocardial tissues of the rats were increased， and the expression of Bax was decreased in the YXF group （,P,<,0.05）. ③ Compared with the YXF group， the CK， LDH， MDA， and H,2,O,2, levels increased and SOD level decreased in the YXF + inhibitor group （,P,<,0.05）. The expressions of Bcl-2， SIRT1， and PGC-1α were decreased and the expression of Bax was increased in the myocardial tissues of the YXF+inhibitor group （,P,<,0.05）.,Conclusion,2,YXF pretreatment can reduce oxidative stress injury and apoptosis and improve MIRI， and its mechanism of action may be related to the regulation of SIRT1/PGC-1α signaling pathway.

关键词

心肌缺血再灌注损伤心肌梗死益心方作用机制大鼠模型中药研究

Keywords

myocardial ischemia-reperfusion injurymyocardial infarctionYixin Formulamechanism of actionrat modeltraditional Chinese herbal medicine research

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