饱和氢盐水对心肌缺血再灌注损伤大鼠冠状动脉内皮细胞功能的影响

Effect of hydrogen-rich saline on coronary artery endothelial cell function during myocardial ischemia-reperfusion in rats

目的探讨饱和氢盐水对心肌缺血再灌注损伤大鼠心肌内皮细胞功能的影响。方法选取健康清洁级雄性SD大鼠150只，体重400-540g。采用随机数字表法将其分为5组（n=30）：空白对照组（Ⅰ组）、假手术组（Ⅱ组）、缺血再灌注组（Ⅲ组）、饱和氢盐水治疗组（Ⅳ组）、生理盐水治疗组（V组）。Ⅰ组不做任何处理；Ⅱ组大鼠仅穿线不结扎；Ⅲ组制备大鼠心肌缺血再灌注损伤模型；Ⅳ组于再灌注前5min给予1ml／100g腹腔注射饱和氢生理盐水；Ⅴ组于再灌注前5min给予1ml／100g腹腔注射等量生理盐水。于再灌注12h、24h后取左心室心肌组织髓染色观察心肌病理学，同时采用电镜观察冠状动脉内皮细胞。采用蛋白质印迹法检测内皮细胞蛋白激酶B（Akt）、内皮型NO合成的限速酶（eNOS）、微血管内皮细胞间黏附分子-1（ICAM-1）和血管细胞黏附分子。1（vCAM-1）。结果与Ⅰ组、Ⅱ组比较，Ⅲ组、Ⅳ组和V组大鼠缺血再灌注12h、24h时内皮细胞Akt、eNOS、ICAM-1和VCAM-1表达水平升高。与III组、V组比较，Ⅳ组大鼠缺血再灌注12h、24h时内皮细胞Akt、eNOS、ICAM-1和VCAM-1表达水平降低。结论饱和氢盐水可改善冠状动脉内皮细胞功能，从而减轻心肌再灌注损伤，其机制可能与抑制Akt／eNOS信号通路，降低ICAM-1、VCAM-1蛋白表达有关。

Objective To investigate the effects of hydrogen-rich saline on coronary artery endothelial cell function during myocardial ischemia-reperfusion in rats. Methods One hundred and fifty healthy male Sprague Dawley rats, weighing 400-540 g were selected. The rats were then randomly divided into 5 groups （n=30）, normal control group （group Ⅰ）; sham operation group （groupⅡ）; myocardial isehemia-reperfusion group （group Ⅲ）; hydrogen-rich saline group （group IV）; normal saline group （group V）. No any processing in group Ⅰ. In group Ⅱ, the anterior descending branch was only exposed but not ligated. Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 rain followed by 12 h, 24 h of reperfusion with Bimbaum. Hydrogen-rich saline 1 ml/100 g were injected intraperitoneally 5 rain before reperfusion in group IV. Normal saline 1 ml/100 g were injected intraperitoneally 5 rain before reperfusion in group V. The rats were sacrificed at 12 h, 24 h of reperfusion and hearts were removed. The pathological changes of myocardial tissue were detected by HE staining. Coronary artery endothelial cells were detected by electron microscope. The expression of Akt, eNOS, ICAM-1 and VCAM-1 in coronary artery endothelial ceils were investigated by Western blot. Results Compared with groups Ⅰ and Ⅱ, the expression of Akt, eNOS, ICAM-1 and VCAM-1 in endothelial cells were significantly increased in groups Ⅲ, Ⅳ and Ⅴ. Compared with group Ⅲ and V, the expression of Akt, eNOS, ICAM-1 and VCAM-1 in endothelial cells were significantly decreased in group Ⅳ. Conclusion Hydrogen-rich saline can reduce myocardial reperfusion injury and improve coronary artery endothelial cell function in rats. The mechanism may be associated with decreasing the expression of Akt, eNOS, ICAM-1 and VCAM-1 in myocardial tissue.