补阳还五汤预处理对心肌缺血再灌注损伤大鼠心肌坏死性凋亡的影响

Effect of Buyang Huanwu Decoction pretreatment on myocardial necrotic apoptosis after myocardial ischemia/reperfusion injury in rats

目的探究补阳还五汤预处理对心肌缺血/再灌注损伤(MI/RI)大鼠心肌坏死性凋亡相关蛋白受体相互作用蛋白激酶1(RIP1)、受体相互作用蛋白激酶3(RIP3)、混合系列蛋白激酶样结构(MLKL)表达的影响,探讨补阳还五汤预处理在MI/RI过程中对坏死性凋亡调控的作用机制。方法将40只SD大鼠随机分为假手术组、模型组、低剂量补阳还五汤组、高剂量补阳还五汤组,每组10只。通过结扎左冠状动脉前降支(LAD)建立大鼠MI/RI模型。补阳还五汤低剂量组和补阳还五汤高剂量组造模前分别给予不同浓度补阳还五汤灌胃,假手术组和模型组给予生理盐水灌胃。通过心电图监测判断造模是否成功,超声心动图检查各组大鼠术后心功能的差异,采用酶联免疫吸附(ELISA)法检测大鼠血清乳酸脱氢酶(LDH)的活性,采用苏木素-伊红染色、原位末端标记法检测大鼠心肌细胞损伤与凋亡,采用蛋白免疫印迹法检测大鼠心脏组织RIP1、RIP3、MLKL表达水平。结果与假手术组比较,模型组大鼠心功能明显降低,心肌细胞炎性浸润明显,心肌细胞凋亡率、RIP1、RIP3、MLKL坏死性凋亡相关蛋白表达均显著升高。与模型组比较,补阳还五汤预处理可改善大鼠心功能,心肌细胞炎性浸润有所改善,心肌细胞凋亡率、RIP1、RIP3、MLKL坏死性凋亡相关蛋白表达均显著降低。此外,补阳还五汤高剂量组较低剂量组改善作用更为显著。结论补阳还五汤能够降低MI/RI大鼠血清LDH活性,抑制炎症反应,改善心脏功能,其作用机制可能与抑制再灌注后心肌细胞的坏死性凋亡有关。

Objective To explore the effects of Buyang Huanwu Decoction pretreatment on the expression of protein receptor-interacting protein kinase 1(RIP1),receptor-interacting protein kinase 3(RIP3)and mixed series protein kinase-like structure(MLKL)in myocardial ischemia/reperfusion injury(MI/RI)rats.To explore the mechanism of regulation of necrotic apoptosis by Buyang Huanwu Decoction pretreatment during MI/RI process.Methods 40 SD rats were randomly divided into sham operation group,model group,low-dose Buyang Huanwu Decoction group and high-dose Buyang Huanwu Decoction group,with 10 rats in each group.Rat MI/RI model was established by ligation of the left anterior descending coronary artery(LAD).Buyang Huanwu decoction low-dose group and Buyang Huanwu Decoction high-dose group were given different concentrations of Buyang Huanwu Decoction by intragastric administration before modeling,sham operation group and model group were given normal saline intragastric administration.ECG monitoring was used to determine whether the modeling was successful,echocardiography was used to examine the differences in postoperative cardiac function of the rats,enzymelinked immunosorbent assay(ELISA)was used to detect the activity of serum lactate dehydrogenase(LDH),hematoxylin-eosin staining and in situ end labeling were used to detect myocardial cell injury and apoptosis.The expression levels of RIP1,RIP3 and MLKL in rat heart tissue were detected by western blot.Results Compared with the sham operation group,the cardiac function of the model group was significantly decreased,the inflammatory infiltration of cardiomyocytes was obvious,and the apoptosis rate of cardiomyocytes,the expression of RIP1,RIP3 and MLKL necrotic apoptosis-related proteins were significantly increased.Compared with the model group,the pretreatment of Buyang Huanwu Decoction could improve the cardiac function of rats,improve the inflammatory infiltration of cardiomyocytes,and significantly reduce the apoptosis rate of cardiomyocytes,the expression of RIP1,RIP3 and MLKL necrotic apoptosation-related proteins.In addition,the improvement effect of Buyang Huanwu Decoction high dose group was more significant than that of low dose group.Conclusion Buyang Huanwu Decoction can reduce serum LDH activity,inhibit inflammatory response and improve cardiac function in MI/RI rats,the mechanism of which may be related to inhibiting the necrotic apoptosis of cardiomyocytes after reperfusion.