乔松素预处理对大鼠心肌缺血再灌注损伤的保护作用研究  

The protective effect of pinocembrin pre-conditioning on rats after myocardial ischemic-reperfusion injury

作　　者：周甄[1] 马梦晴 林先和[2] 刘和俊[2] 

机构地区：[1]安徽医科大学第二临床医学院,合肥230032 [2]安徽医科大学第一附属医院心血管内科,合肥230022

出　　处：《安徽医科大学学报》2015年第12期1729-1733,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1508085MH145)

摘　　要：目的探讨不同浓度的乔松素预处理对大鼠心肌缺血再灌注损伤(IRI)的保护作用及其相关机制。方法成年SD大鼠125只,45只用于前期筛选乔松素适宜用药浓度,另80只随机均分为假手术组,模型组,乔松素低、中、高剂量(3、10、30 mg/kg)组。通过结扎大鼠心左冠状动脉前降支30 min并解开结扎线释放血流2 h建立大鼠在体IRI模型,乔松素组于缺血前1 h尾静脉给药,模型建立后HE染色观察心肌组织形态学改变;免疫荧光染色法检测自噬标志物LC3的表达及定位;TUNEL原位标记凋亡心肌细胞;Western blot法检测凋亡蛋白Bax、Caspase-3及自噬蛋白LC3、p62的表达。结果 HE染色结果显示,合适浓度的乔松素能有效改善心肌组织病理学改变,且呈剂量依赖性,但剂量较高时可能具有毒性作用;免疫荧光染色显示乔松素组LC3表达明显较模型组增多,在胞质中广泛表达,且呈剂量依赖性;TUNEL染色结果显示,乔松素组与模型组比较,凋亡细胞均明显减少,且高剂量组优于中、低剂量组;Western blot结果显示,乔松素组Bax、Caspase-3、p62蛋白表达水平较模型组明显降低(P<0.05),且高剂量组效果更佳(P<0.01);乔松素组LC3表达水平高于模型组(P<0.05),且高剂量组增高优于中、低剂量组(P<0.01)。结论乔松素预处理对大鼠心肌IRI有保护作用,其主要作用机制可能与减少细胞凋亡和激活自噬有关。Objective To investigate the protective effect and its mechanism of pinocembrin pre - conditioning on rats after myocardial ischemic - reperfusion injury （IRI）. Methods 45 male sprague - dawley rats were used for exploring the suitable concentration of the pinocembrin and 80 rats were equally divided into 5 groups ： sham group, model group, low, middle, high dose （3, 10, 30 mg/kg） group. The IRI models were set up by ligating ratg left anterior descending coronary artery for 30 minutes then loosing it for 2 hours. Pinocembrin was gave 1 hour via tail intravenous injection before ischemic - reperfusion. The morphological changes of myocardial tissues were observed by HE staining. The LC3 expression and position were measured by immunofluorescence. The apoptosis of myocardial ceils was measured by TUNEL staining. The expression of apoptosis - related protein such as Bax, Caspase-3, p62 and autophagy-related protein,LC3 were detected by Western blot. Results HE staining showed that the path- ological changes of myocardial tissues were improved by pinocembrin and presented a dose-dependence. However, high-dose of drug might bring the drug toxicity. The expression of LC3 was more in the test group than the model group as evidenced by immunofluorescence staining and in a dose dependent fashion. Compared with the model group, the TUNEL-labeled myocardial cells were significantly decreased in pinocembrin groups, and the level of apoptotic cells were negatively related to the pinocembrin concentration. Western blot results showed that Bax, Caspase-3 and p62 in pinocembrin groups were significantly lower than in model group, The levels of apoptotic pro- teins were negatively related to the pinocembrin concentration. In reverse, compared with the model group, the au- tophagic-related protein, LC3, showed an increasing trend in the pinocembrin groups and presented a positive rela- tion to drug＇s concentration. Conclusion The results indicate that the pinocembrin pre -conditioning shows the protective effect on ra

关 键 词：乔松素 心肌缺血再灌注 细胞凋亡 自噬 

分 类 号：R542.2[医药卫生—心血管疾病] R932[医药卫生—内科学]