人参皂甙Re对大鼠急性缺血再灌注心肌细胞凋亡及相关基因蛋白表达的影响(英文)

Effects of ginsenoside-Re on anti-cardiomyocyte apoptosis and the expression of related gene proteins in rats after acute ischemia-reperfusion

背景:心肌细胞缺血再灌注损伤近来已成为临床研究的热点,但人参及其提取物对急性缺血再灌注损伤后心肌细胞凋亡的作用尚未阐明。目的:观察人参皂甙Re对急性缺血再灌注心肌细胞凋亡及Bcl-2,Bax,Bad和Fas基因蛋白表达的影响,探讨人参皂甙Re抑制心肌细胞凋亡的可能机制。设计:随机对照实验研究。地点、对象和干预:本实验在湖北省武汉市同济医院动物实验中心完成。实验共用Wistar大鼠15只,雌雄不分。按随机数字表法分为假手术组、缺血再灌注组、人参皂甙Re治疗组,每组各5只。建立Wistar鼠缺血再灌注心脏模型。应用原位末端标记法检测心肌凋亡细胞,并进行细胞计数;原位杂交和免疫组化检测Bcl-2mRNA和基因表达产物的蛋白质,通过图像分析系统测量阳性染色区域的平均吸光度值进行量化分析,以观察人参对大鼠心肌缺血再灌注心肌细胞凋亡的影响及基因表达。主要观察指标:人参皂甙Re治疗与否对急性大鼠缺血再灌注心肌细胞凋亡及Bcl-2,Bax,Bad和Fas基因蛋白表达的影响。结果:①假手术组未发现心肌凋亡细胞。缺血再灌注组心肌凋亡细胞数为(134±46)个/视野,人参皂甙Re治疗组为(91±19)个/视野,两组间差异有显著性意义(t=4.63,P<0.01)。②免疫组织化学检测发现缺血再灌注组及人参皂甙Re治疗组Bcl-2,Bax,Bad和Fas基因蛋?

BACKGROUND:Myocardial ischemia and reperfusion injury has become the clinical hotspot for research recently.Whether Ginseng and its extract ginsenoride have the effect on the cardiomyocyte apoptosis induced by acute ischemia reperfusion injury is not clear. OBJECTIVE:To observe the effects of ginsenoside Re on anti cardiomyocyte apoptosis and the expression of Bcl 2,Bax,Bad and Fas proteins in rats after acute ischemia and reperfusion,and explore the possible mechanisms of ginsenoside Re on the inhibition of cardiomyocyte apoptosis. DESIGN:Randomized and controlled experimental study. SETTING,PARTICIPANTS and INTERVENTION:Fifteen Wistar rats(provided by Zoology Department of Tongji Medical University) of both sexes,weighing 200-500 g were used in the present study.The research was conducted in the Experimental Center of Tongji Hospital,Wuhan,Hubei Province,China.The heart model of acute ischemia reperfusion was established by ligating the left anterior descending branch of coronary artery in Wistar rats.The apoptotic cardiomyocytes were measured with transmission electron microscopy and counted with in situ nick end labeling(TUNEL) method and optic microscopy.The expression of Bcl 2,Bax,Bad and Fas proteins were studied by immunohistochemical staining.Mean optical density(OD) value of the expression of the proteins was quantitatively examined by image analysis system. MAIN OUTCOME MEASURES:Whether ginsenoside Re has the effects on cardiomyocyte apoptosis and the influence of the expression of Bcl 2,Bax,Bad,and Fas proteins in rats after acute ischemia reperfusion. RESULTS:①The apoptotic cardiomyocytes did not exist in the sham operation group.The number of the apoptotic cells was(134.5±45.61)/field in the ischemia/reperfusion(I/R) group,and(90.66±19.22)/field in the ginsenodside Re treated group,and the difference was significant between the latter two groups(t=4.63,P< 0.01).②The expression of Bcl 2,Bax,Bad and Fas proteins increased significantly in the I/R group and ginsenodside Re treated group,compared with the sham operation group(t=2.79,P< 0.05).There was no significant difference in the Bcl 2 protein expression between the ginsenodside Re treated group and I/R group(t=2.67,P >0.05),whereas the expression of Bax,Bad,and Fas genes were decreased significantly in the ginsenodside Re treated group,compared with the I/R group(t=2.81,P< 0.05).Compared with the I/R group and the sham operation group,the ratios of Bcl 2/Bax,Bcl 2/Bad,and Bcl 2/Fas were increased significantly in the ginsenodside Re treated group(t=2.84,P< 0.05). CONCLUSION:Ginsenoside Re can significantly inhibit cardiomyocyte apoptosis induced by acute ischemia/reperfusion injury in rats.The mechanism is that ginsenoside Re inhibits the expression of pro apoptotic bax,bad and fas gene and raises the ratios of bcl 2/bax,bcl 2/bad,and bcl 2/fas.