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[乌附经方预处理对大鼠缺血/再灌注损伤诱导凋亡的影响]

[Effects of preconditioning of wufu jingfang on ischemia/reperfusion injury induced apoptosis in rats].

作者信息

Li Ling, Ma Yu-Hong, Wang Su-Qing

机构信息

School of Public Health, Wuhan University, Wuhan 430072, China.

出版信息

Zhongguo Zhong Xi Yi Jie He Za Zhi. 2013 Jun;33(6):789-94.

Abstract

OBJECTIVE

To observe the protective effect of Wufu Jingfang (WJ, containing Aconitum carmichaeli Debx, Radix Aconiti Lateralis Preparatae, Rhizoma Pinelliae, and snakegourd fruit) on myocardial ischemia-reperfusion injury (I/R) of rats, thus exploring the feasibility of recipes containing eighteen incompatible pairs for specific pathological conditions.

METHODS

Fifty male Wistar rats were randomly divided into five groups, i.e., the sham-operative control group (the SH group), the I/R group, the low dose WJ I/R group (the I/R +JFL group), the middle dose WJ I/R group (the I/R +JFM group), the high dose WJ I/R group (the I/R +JFH group), 10 in each group. Rats in the latter three groups were administered with WJ at 0.75 mL/100 g, 1.50 mL/100 g, and 3.00 mL/100 g body weight for 14 consecutive days by gastrogavage. All groups except the SH group received ligation of left anterior descending branch of coronary artery for 30-min ischemia followed by 120-min reperfusion. The micro-structural changes of myocardial mitochondria were observed by transmission electron microscope. The ischemic cardiomyocyte apoptosis was detected in each group using one-step terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mRNA expressions of B-cell leukemia/lymphoma 2 (Bcl-2) and Bcl-2 associated x protein (Bax) were detected by RT-PCR. The activities of lactic dehydrogenase (LDH) and creatine kinase (CK) were detected using ELISA. The myocardial infarct size was detected.

RESULTS

Compared with the I/R group, WJ pretreatment significantly suppressed the release of LDH and CK (Besides, the release of LDH and CK reduced along with increased dose.), reduced the myocardial infarct size, and lowered myocardial apoptosis index (P < 0.05). WJ pretreatment also modulated Bcl-2/Bax ratio by up-regulating Bcl-2 expression level while decreasing Bax expression level.

CONCLUSIONS

WJ pretreatment might protect the heart from I/R injury via decreasing myocardial cell apoptosis. The results suggested that eighteen incompatible pairs is not absolute, but relative. Chinese medical preparation containing opposite Chinese herbs could be used in specific pathological states such as ischemic cardiomyopathy.

摘要

目的

观察五福经方(由制川乌、制附子、半夏、瓜蒌组成)对大鼠心肌缺血再灌注损伤的保护作用,探讨十八反配伍组方用于特定病理状态的可行性。

方法

将50只雄性Wistar大鼠随机分为5组,即假手术对照组(SH组)、缺血再灌注组(I/R组)、五福经方低剂量缺血再灌注组(I/R+JFL组)、五福经方中剂量缺血再灌注组(I/R+JFM组)、五福经方高剂量缺血再灌注组(I/R+JFH组),每组10只。后三组大鼠分别按0.75 mL/100 g、1.50 mL/100 g、3.00 mL/100 g体重灌胃给予五福经方,连续14天。除SH组外,其余各组均采用结扎冠状动脉左前降支的方法,造成心肌缺血30分钟,再灌注120分钟。用透射电镜观察心肌线粒体超微结构变化;采用原位末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)检测各组缺血心肌细胞凋亡情况;用逆转录聚合酶链反应(RT-PCR)检测B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的mRNA表达;用酶联免疫吸附测定法(ELISA)检测乳酸脱氢酶(LDH)和肌酸激酶(CK)活性;测定心肌梗死面积。

结果

与I/R组比较,五福经方预处理能显著抑制LDH和CK的释放(且随着剂量增加,LDH和CK的释放量减少),缩小心肌梗死面积,降低心肌凋亡指数(P<0.05)。五福经方预处理还可通过上调Bcl-2表达水平、下调Bax表达水平来调节Bcl-2/Bax比值。

结论

五福经方预处理可能通过减少心肌细胞凋亡对心脏起到缺血再灌注损伤保护作用。结果提示十八反并非绝对,而是相对的。含相反中药的方剂在缺血性心肌病等特定病理状态下可以应用。

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