加味丹参饮作用内源性H2S合成途径保护心肌缺血/再灌注损伤的实验研究

Experimental Study on Effects of Jiawei Danshen Decoction on Endogenous H2S Synthesis Pathway to Protect Myocardial Ischemia/Reperfusion Injury

目的通过观察益气活血中药组方加味丹参饮(JWDS)对心肌缺血/再灌注损伤(myocardial ischemia reperfusion injury,MIRI)模型大鼠血清硫化氢(H2S)合成酶/胱硫醚-γ-裂解酶(cystathionine-γ-lyse,CSE)、肌酸激酶同工酶(creatine kinase isoenzymes,CK)、乳酸脱氢酶(lactate dehydrogenase,DH)含量、心肌组织超微结构、心肌组织CSEmRNA表达的影响,从内源性H2S合成途径探讨JWDS治疗MIRI的作用机制。方法给药组大鼠按剂量6.19g/(kg·d)要求给药14d,末次给药2h后采用结扎冠脉左前降支/再灌流方法复制MIRI模型。HE染色观察心肌组织机构改变情况;ELISA法检测血清CK、LDH、CSE含量;Western-blot检测心肌组织CSE蛋白表达;Real-timePCR检测心肌组织CSEmRNA表达。结果JWDS可明显改善MIRI模型大鼠心肌组织病理改变,降低血清LDH、CK含量(P<0.01),升高CSE含量(P<0.01),上调心肌组织CSE及mRNA表达(P<0.05,P<0.01)。PPG可明显降低JWDS组大鼠血清CSE含量(P<0.01),下调心肌CSEmRNA表达(P<0.01)。结论JWDS对实验性心肌缺血/再灌注(MIR)大鼠心肌损伤具有明显保护作用,其机制与促进内源性H2S生成从而保护心肌细胞结构,抑制CK、LDH漏出,上调心肌组织CSE蛋白及mRNA表达有关。

Objective To observe effects of a prescription for strengthening Qi and promoting blood-Jiawei Danshen Decoction (JWDS) on serum contents of H2S synthetase/cystathionine-γ-lyase (CSE), creatine kinase isoenzyme (CK) and lactate dehydrogenase (LDH), myocardial tissue ultrastructure, and CSE mRNA expression in myocardial tissue of model rats with myocardial ischemia reperfusion injury (MIRI), so as to explore the mechanism of JWDS in treating MIRI through synthesis pathway of endogenous H2S. Methods Rats in medication group were given medicine 6.19 g/(kg·d) for 14 d, and 2 h after the last medication, the MIRI model was replicated by ligation of left anterior descending coronary artery/reperfusion method. Ultrastructural changes of myocardial tissue were observed by HE staining. The contents of serum CK, LDH and CSE were detected by ELISA method. The expression of CSE protein in myocardial tissue was detected by Western-blot. The expression of CSE mRNA in myocardial tissue was detected by Real-time PCR. Results JWDS significantly improved pathological changes of myocardial tissue in model rats with MIRI, decreased serum LDH and CK contents (P<0.01), increased CSE content (P<0.01), and up-regulated CSE and mRNA expression in myocardial tissue (P<0.05, P<0.01). PPG significantly decreased serum CSE content in rats of the JWDS group (P<0.01), and down-regulated CSE mRNA expression in myocardial tissue (P<0.01). Conclusion JWDS demonstrates significant protective effect on myocardial injury in experimental MIRI model rats, which is associated with promoting endogenous H2S generation to protect myocardial cell structure, inhibit CK and LDH leakage, and up-regulate CSE protein and mRNA expression in myocardial tissue.