依托咪酯后处理对大鼠肝缺血再灌注肾细胞凋亡及Bcl-2和Bax表达的影响
发布时间：2018-11-17 19:17
【摘要】：目的:探讨依托咪酯后处理对大鼠肝缺血再灌注后肾损伤以及Bcl-2、Bax蛋白表达的影响。方法:健康雄性Wistar大鼠54只随机分为3组:假手术组(Sham组)、缺血再灌注组(I/R组)、依托咪酯组(Eto组)。每组又根据不同的再灌注时间(2、4、6h)分为3个时相。除Sham组之外其余各组均为肝脏缺血1h以后恢复再灌注,Sham组和I/R组在缺血50min时间点开始持续静脉泵注0.9%氯化钠注射液0.1ml·kg-1·min-1,Eto组持续静脉泵注依托咪酯脂肪乳1.0ml(0.3mg·kg-1稀释到1.0ml),两组泵注时间均为10min。实验结束时即刻取左肾下极组织,行组织切片以及苏木精-伊红(HE)染色,然后在光镜下观察不同大鼠肝缺血再灌注时间肾组织的病理切片。术毕心脏取血测定血浆胱抑素C(Cysc)的表达;采用免疫组化方法检测肾组织抑癌基因Bcl-2蛋白与促癌基因Bax蛋白表达量以及肾细胞凋亡指数(AI)。结果:(1)TUNEL检测结果:与Sham组比较,I/R组和Eto组肾组织AI值均增高(P0.05);与I/R组比较,Eto组肾组织的AI值下调(P0.05);且在同一再灌注时间时,Sham组、Eto组、I/R组的AI值依次升高,并以I/R组再灌注6h时最高。(2)与Sham组比较,I/R组和Eto组Bcl-2和Bax表达均上调(P0.05);与I/R组比较,Eto组Bcl-2蛋白表达增多、Bax蛋白表达减少(P0.05)。且在同一再灌注时间时,Sham组、I/R组、Eto组的Bcl-2蛋白值依次升高,并以Eto组再灌注6h时最高;Sham组、Eto组、I/R组的Bax蛋白值依次升高,并以I/R组再灌注6h时最高。(3)与Sham组比较,I/R组和Eto组Cysc含量均升高(P0.05);与I/R组比较,Eto组Cysc含量下降(P0.05)。且在同一再灌注时间时,Sham组、Eto组、I/R组的Cysc含量升高,并以I/R组再灌注6h时最高。(4)光镜下I/R组的肾组织病理学的变化比Eto组明显。结论:依托咪酯后处理可以减轻肝缺血再灌注后肾组织的损伤,其机制可能是通过使抑癌基因Bcl-2蛋白的表达增多,促癌基因Bax蛋白的表达减少来发挥对肾组织细胞的保护作用。
[Abstract]:Aim: to investigate the effects of etomidate postconditioning on renal injury and expression of Bcl-2,Bax protein after hepatic ischemia reperfusion in rats. Methods: 54 healthy male Wistar rats were randomly divided into three groups: sham operation group (Sham group), ischemia reperfusion group (I / R group) and etomidate group (Eto group). Each group was divided into 3 phases according to different reperfusion time (2? 4? 6 hours). All the other groups except the Sham group resumed reperfusion after 1 hour of hepatic ischemia. The Sham group and the I / R group began to continuously inject 0.9% sodium chloride 0.1ml kg-1 min-1, at the time point of ischemia 50min. Eto group received continuous intravenous injection of etomidate fat emulsion 1.0ml (0.3mg kg-1 diluted to 1.0ml) for 10 min. Tissue sections and hematoxylin-eosin (HE) staining were performed immediately at the end of the experiment. Then the pathological sections of renal tissues were observed under light microscope at different time of hepatic ischemia reperfusion. The expression of cystatin C (Cysc) in plasma and renal tissue tumor suppressor gene Bcl-2 protein and promoting gene Bax protein expression and renal cell apoptosis index (AI).) were detected by immunohistochemical method. Results: (1) TUNEL: compared with Sham group, the AI value of renal tissue in I / R group and Eto group was increased (P0.05), and the AI value of renal tissue in Eto group was down-regulated (P0.05) compared with I / R group. At the same time of reperfusion, the AI values of Sham group, Eto group and I / R group increased in turn, and the highest at 6 h after reperfusion in I / R group. (2) compared with Sham group, Bcl-2 and Bax expression in I / R group and Eto group were all up-regulated (P0.05). Compared with I / R group, Bcl-2 protein expression increased and Bax protein expression decreased in Eto group (P0.05). At the same time of reperfusion, the value of Bcl-2 protein in Sham group, I / R group and Eto group increased in turn, and the highest value was in Eto group at 6 h after reperfusion. The value of Bax protein in Sham group, Eto group and I / R group increased in turn, and the highest value was found at 6 h after reperfusion in I / R group. (3) compared with Sham group, Cysc content in I / R group and Eto group increased significantly (P0.05). Compared with I / R group, Cysc content in Eto group decreased (P0.05). At the same time of reperfusion, the content of Cysc increased in Sham group, Eto group and I / R group, and the highest at 6 h after reperfusion in I / R group. (4) the pathological changes of renal tissue in I / R group were significantly higher than those in Eto group under light microscope. Conclusion: etomidate post-treatment can attenuate the injury of renal tissue after hepatic ischemia-reperfusion, and the mechanism may be by increasing the expression of tumor suppressor gene Bcl-2 protein. The expression of oncogene Bax protein was reduced to protect renal tissue cells.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R614
本文编号：2338801
[Abstract]:Aim: to investigate the effects of etomidate postconditioning on renal injury and expression of Bcl-2,Bax protein after hepatic ischemia reperfusion in rats. Methods: 54 healthy male Wistar rats were randomly divided into three groups: sham operation group (Sham group), ischemia reperfusion group (I / R group) and etomidate group (Eto group). Each group was divided into 3 phases according to different reperfusion time (2? 4? 6 hours). All the other groups except the Sham group resumed reperfusion after 1 hour of hepatic ischemia. The Sham group and the I / R group began to continuously inject 0.9% sodium chloride 0.1ml kg-1 min-1, at the time point of ischemia 50min. Eto group received continuous intravenous injection of etomidate fat emulsion 1.0ml (0.3mg kg-1 diluted to 1.0ml) for 10 min. Tissue sections and hematoxylin-eosin (HE) staining were performed immediately at the end of the experiment. Then the pathological sections of renal tissues were observed under light microscope at different time of hepatic ischemia reperfusion. The expression of cystatin C (Cysc) in plasma and renal tissue tumor suppressor gene Bcl-2 protein and promoting gene Bax protein expression and renal cell apoptosis index (AI).) were detected by immunohistochemical method. Results: (1) TUNEL: compared with Sham group, the AI value of renal tissue in I / R group and Eto group was increased (P0.05), and the AI value of renal tissue in Eto group was down-regulated (P0.05) compared with I / R group. At the same time of reperfusion, the AI values of Sham group, Eto group and I / R group increased in turn, and the highest at 6 h after reperfusion in I / R group. (2) compared with Sham group, Bcl-2 and Bax expression in I / R group and Eto group were all up-regulated (P0.05). Compared with I / R group, Bcl-2 protein expression increased and Bax protein expression decreased in Eto group (P0.05). At the same time of reperfusion, the value of Bcl-2 protein in Sham group, I / R group and Eto group increased in turn, and the highest value was in Eto group at 6 h after reperfusion. The value of Bax protein in Sham group, Eto group and I / R group increased in turn, and the highest value was found at 6 h after reperfusion in I / R group. (3) compared with Sham group, Cysc content in I / R group and Eto group increased significantly (P0.05). Compared with I / R group, Cysc content in Eto group decreased (P0.05). At the same time of reperfusion, the content of Cysc increased in Sham group, Eto group and I / R group, and the highest at 6 h after reperfusion in I / R group. (4) the pathological changes of renal tissue in I / R group were significantly higher than those in Eto group under light microscope. Conclusion: etomidate post-treatment can attenuate the injury of renal tissue after hepatic ischemia-reperfusion, and the mechanism may be by increasing the expression of tumor suppressor gene Bcl-2 protein. The expression of oncogene Bax protein was reduced to protect renal tissue cells.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R614
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