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背景:肾移植中肾缺血再灌注损伤能引发凋亡程序的执行,丙泊酚可能的肾保护作用及其与细胞凋亡通路间的关系,有助于探讨丙泊酚的作用机制。目的:探讨丙泊酚对大鼠肾缺血再灌注损伤细胞凋亡通路的影响及可能的作用机制。设计、时间及地点:动物实验,细胞形态学观察,于2004/2005在北京友谊医院泌尿外科研究所实验室共同设计和完成。材料:选取封闭群SD成年雄性大鼠99只。兔抗鼠Bcl-2、Bax、caspase3、cytochromeC均为武汉博士德生物工程有限公司产品。方法:将动物随机分为对照组26只、缺血再灌注组35只、缺血再灌注+丙泊酚组38只。建立大鼠肾缺血再灌注损伤模型,术前单次静脉缓慢输注乳酸林格液5mL/kg,逐层正中切口开腹,切除右肾,用无创血管夹夹闭左侧肾蒂,缺血时间45min,松夹再灌注后全层缝合腹壁,取再灌注0,3,12,24,72h左侧肾脏,取肾同时采血处死动物。缺血再灌注+丙泊酚组在缺血前15min用药,丙泊酚1mg/(kg?min)直至再灌注后30min,共用药75min。对照组和缺血再灌注组以乳酸林格液等量输注。主要观察指标:观察损伤肾的形态学改变,用免疫组织化学观察细胞凋亡相关蛋白Bcl-2、Bax、caspase3和细胞色素C的表达情况。结果:光镜可观察到肾缺血再灌注引起的肾损害,程度依次为近曲小管、远曲小管、集合管、肾小球;免疫组化图像分析观察到缺血再灌注组与对照组相比,Bax、caspase3,细胞色素C表达增加,Bcl-2表达未见明显变化;缺血再灌注+丙泊酚组与对照组比较,前者Bax、caspase3和细胞色素C表达下降,Bcl-2表达增高(P<0.05)。结论:丙泊酚对肾缺血再灌注损伤引起的细胞凋亡可能具有保护作用,可能机制是抑制促凋亡蛋白Bax、caspase3和细胞色素C的表达,对Bcl-2的表达无影响,与细胞凋亡的线粒体通路途径有关。
BACKGROUND: Renal ischemia-reperfusion injury during renal transplantation leads to the execution of apoptosis programs, the possible renal protective effect of propofol and its relationship with apoptotic pathways, which may help to explore the mechanism of propofol. Objective: To investigate the effect of propofol on apoptosis pathway of renal ischemia-reperfusion injury in rats and its possible mechanism. DESIGN, TIME AND SETTING: Animal experiments and observation of cell morphology were jointly designed and completed at the Laboratory of Urology, Beijing Friendship Hospital, 2004/2005. MATERIALS: Thirty-nine SD adult male rats were selected. Rabbit anti-mouse Bcl-2, Bax, caspase3, cytochromeC are Wuhan Boster Biological Engineering Co., Ltd. products. Methods: The animals were randomly divided into control group (n = 26), ischemia - reperfusion group (n = 35) and ischemia / reperfusion plus propofol group (n = 38). A rat model of renal ischemia-reperfusion injury was established. A single intravenous infusion of lactic acid Ringer’s solution (5 mL / kg) was performed preoperatively. The right middle kidney was excised one by one and the left renal pedicle was closed with a noninvasive vascular clamp. The blood time was 45 minutes. After the pinecone reperfusion, the whole abdominal wall was sutured, and the left kidney at 0, 3, 12, 24 and 72 hours after reperfusion was sacrificed. Ischemia reperfusion + propofol group 15min before ischemia, propofol 1mg / (kg? Min) until 30min after reperfusion, a total of 75min. The control group and ischemia-reperfusion group were injected with lactated Ringer’s solution in equal volume. MAIN OUTCOME MEASURES: The morphological changes of injured kidneys were observed. The expressions of Bcl-2, Bax, caspase3 and cytochrome C were observed by immunohistochemistry. Results: Renal ischemia-reperfusion-induced renal damage was observed by light microscopy, followed by proximal convoluted tubule, distal convoluted tubule, collecting duct and glomeruli. Immunohistochemical analysis showed that ischemia reperfusion group and control group Compared with the control group, the expressions of Bax, caspase3 and cytochrome C were decreased in the ischemia-reperfusion + propofol group and the expressions of Bax, caspase3 and cytochrome C were up-regulated while the expression of Bcl-2 The expression was increased (P <0.05). CONCLUSION: Propofol may have a protective effect on apoptosis induced by renal ischemia / reperfusion injury. The possible mechanism is to inhibit the expression of pro-apoptotic proteins Bax, caspase 3 and cytochrome C, but not Bcl-2 Mitochondrial pathway of apoptosis related.