【摘要】：目的:探討依托咪酯后處理對大鼠肝缺血再灌注后腎損傷以及Bcl-2、Bax蛋白表達的影響。方法:健康雄性Wistar大鼠54只隨機分為3組:假手術(shù)組(Sham組)、缺血再灌注組(I/R組)、依托咪酯組(Eto組)。每組又根據(jù)不同的再灌注時間(2、4、6h)分為3個時相。除Sham組之外其余各組均為肝臟缺血1h以后恢復(fù)再灌注,Sham組和I/R組在缺血50min時間點開始持續(xù)靜脈泵注0.9%氯化鈉注射液0.1ml·kg-1·min-1,Eto組持續(xù)靜脈泵注依托咪酯脂肪乳1.0ml(0.3mg·kg-1稀釋到1.0ml),兩組泵注時間均為10min。實驗結(jié)束時即刻取左腎下極組織,行組織切片以及蘇木精-伊紅(HE)染色,然后在光鏡下觀察不同大鼠肝缺血再灌注時間腎組織的病理切片。術(shù)畢心臟取血測定血漿胱抑素C(Cysc)的表達;采用免疫組化方法檢測腎組織抑癌基因Bcl-2蛋白與促癌基因Bax蛋白表達量以及腎細胞凋亡指數(shù)(AI)。結(jié)果:(1)TUNEL檢測結(jié)果:與Sham組比較,I/R組和Eto組腎組織AI值均增高(P0.05);與I/R組比較,Eto組腎組織的AI值下調(diào)(P0.05);且在同一再灌注時間時,Sham組、Eto組、I/R組的AI值依次升高,并以I/R組再灌注6h時最高。(2)與Sham組比較,I/R組和Eto組Bcl-2和Bax表達均上調(diào)(P0.05);與I/R組比較,Eto組Bcl-2蛋白表達增多、Bax蛋白表達減少(P0.05)。且在同一再灌注時間時,Sham組、I/R組、Eto組的Bcl-2蛋白值依次升高,并以Eto組再灌注6h時最高;Sham組、Eto組、I/R組的Bax蛋白值依次升高,并以I/R組再灌注6h時最高。(3)與Sham組比較,I/R組和Eto組Cysc含量均升高(P0.05);與I/R組比較,Eto組Cysc含量下降(P0.05)。且在同一再灌注時間時,Sham組、Eto組、I/R組的Cysc含量升高,并以I/R組再灌注6h時最高。(4)光鏡下I/R組的腎組織病理學(xué)的變化比Eto組明顯。結(jié)論:依托咪酯后處理可以減輕肝缺血再灌注后腎組織的損傷,其機制可能是通過使抑癌基因Bcl-2蛋白的表達增多,促癌基因Bax蛋白的表達減少來發(fā)揮對腎組織細胞的保護作用。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R614

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