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中药联合缺血后适应对大鼠心肌缺血再灌注损伤的保护机制

[Mechanisms of Chinese herbs combined with ischemic postconditioning in protecting myocardium of rats from ischemia-reperfusion injury].

作者信息

Zhang Da-wu, Zhang Lei, Liu Jian-gang, Wang Cheng-long, Shi Da-zhuo, Chen Ke-ji

机构信息

Department of Cardiology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China.

出版信息

Zhong Xi Yi Jie He Xue Bao. 2010 May;8(5):465-71.

Abstract

OBJECTIVE

To observe the effects of Chinese patent medicines with the function of replenishing qi to activate blood (RQAB) plus ischemic postconditioning (IPoC) in protecting myocardium of rats from ischemia-reperfusion (I/R) injury, and to explore the possible mechanisms.

METHODS

Seventy-five Sprague-Dawley rats were randomly divided into sham-operated group (the suture was penetrated around the left anterior descending coronary artery, but not tied, n=15), I/R group (30 minutes of in situ transient occlusion of the left anterior descending artery, followed by 1 hour of reperfusion, n=15), IPoC group (30 minutes occlusion of the left anterior descending artery, followed by 3 cycles of 10 s of reperfusion/10 s of ischemia before 1-hour reperfusion, n=15), RQAB plus IPoC group (pretreated with 0.162 g/kg Xinyue Capsule and 0.135 g/kg Xiongshao Capsule for 14 days, and treated with IPoC 2 h after the final gavage, n=15), fosinopril sodium plus IPoC group (pretreated with fosinopril sodium, 0.9 mg/kg, n=15). Serum creatine kinase-MB (CK-MB) activity and cardiac troponin T (cTnT) level were detected; myocardial infarction size was measured by nitrotetrazolium blue chloride staining; Toll-like receptor 2 (TLR2) and TLR4 in myocardial tissue were examined by immunohistochemical method; interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) levels in myocardial tissue were examined by enzyme-linked immunosorbant assay.

RESULTS

Compared with the I/R group, myocardial enzymes and infarction size were decreased significantly in the IPoC group (P<0.01); expressions of TLR2 and TLR4 and levels of IL-1beta and IL-6 in myocardial tissues were also significantly lower than those in the I/R group (P<0.05). Compared with the fosinopril sodium plus IPoC group, expressions of TLR2 and TLR4 were decreased significantly in the RQAB plus IPoC group (P<0.05, P<0.01). Compared with IPoC, RQAB plus IPoC reduced the infarction size and the release of myocardial enzyme CK-MB (P<0.01), and decreased the expressions of TLR2 and TLR4 and the levels of IL-1beta and IL-6 (P<0.05, P<0.01) in myocardial tissues.

CONCLUSION

Pretreatment with Chinese herbs for nourishing qi and activating blood circulation can enhance the protective effect of IPoC on rat myocardial I/R injury, and its mechanism may be related to inhibition of TLR expression and expressions of the downstream proinflammatory cytokines.

摘要

目的

观察益气活血类中成药联合缺血后处理(IPoC)对大鼠心肌缺血再灌注(I/R)损伤的保护作用,并探讨其可能机制。

方法

将75只Sprague-Dawley大鼠随机分为假手术组(在左冠状动脉前降支周围穿线,但不结扎,n=15)、I/R组(左前降支原位短暂阻断30分钟,随后再灌注1小时,n=15)、IPoC组(左前降支阻断30分钟,在1小时再灌注前进行3个循环的10秒再灌注/10秒缺血,n=15)、益气活血联合IPoC组(用0.162 g/kg心悦胶囊和0.135 g/kg芎芍胶囊预处理14天,末次灌胃后2小时进行IPoC处理,n=15)、福辛普利钠联合IPoC组(用0.9 mg/kg福辛普利钠预处理,n=15)。检测血清肌酸激酶同工酶(CK-MB)活性和心肌肌钙蛋白T(cTnT)水平;用氯化硝基四氮唑蓝染色法测定心肌梗死面积;采用免疫组织化学方法检测心肌组织中Toll样受体2(TLR2)和TLR4;采用酶联免疫吸附测定法检测心肌组织中白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)水平。

结果

与I/R组比较,IPoC组心肌酶和梗死面积明显降低(P<0.01);心肌组织中TLR2和TLR4表达以及IL-1β和IL-6水平也明显低于I/R组(P<0.05)。与福辛普利钠联合IPoC组比较,益气活血联合IPoC组TLR2和TLR4表达明显降低(P<0.05,P<0.01)。与IPoC组比较,益气活血联合IPoC组梗死面积和心肌酶CK-MB释放减少(P<0.01),心肌组织中TLR2和TLR4表达以及IL-1β和IL-6水平降低(P<0.05,P<0.01)。

结论

益气活血类中药预处理可增强IPoC对大鼠心肌I/R损伤的保护作用,其机制可能与抑制TLR表达及下游促炎细胞因子表达有关。

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