二甲双胍预处理对大鼠肾缺血再灌注损伤模型的影响及机制研究
投稿时间:2017-01-09  修订日期:2017-01-11  点此下载全文
引用本文:柳永,刘修恒.二甲双胍预处理对大鼠肾缺血再灌注损伤模型的影响及机制研究[J].医学研究杂志,2017,46(9):65-68
DOI: 10.11969/j.issn.1673-548X.2017.09.018
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作者单位E-mail
柳永 430060 武汉大学人民医院泌尿外科  
刘修恒 430060 武汉大学人民医院泌尿外科 944047761@qq.com 
基金项目:湖北省首届领军人才培养工程培养对象(湖北省卫生与计划生育委员会);武汉市应用基础研究计划项目(2015060101010049)
中文摘要:目的 探讨二甲双胍预处理对大鼠肾缺血再灌注损伤模型的影响及可能的机制。方法 健康清洁级成年雄性SD大鼠体重220~250g,共30只,采用数字表法将其随机分为3组(n=10):假手术组(Sham组),缺血再灌注组(I/R组)和二甲双胍组(metformin+I/R组)。metformin+I/R组在建立I/R模型前给予二甲双胍0.125mg/(kg·d)腹腔注射,共14天,Sham组和I/R组给予等容量生理盐水连续14天。给药结束后I/R组和metformin+I/R组采用切除右肾、夹闭左肾动静脉45min后恢复灌注的方法制备大鼠肾缺血再灌注模型,Sham组切除右肾、不夹闭左肾动静脉。分别于再灌注24h时抽取下腔静脉血测定血清肌酐(Cr)和尿素氮(BUN)浓度;然后处死大鼠取肾,光镜下观察肾组织形态学变化,免疫组化法测定Bcl-2、Bax、caspase-3表达,并行Tunel染色评估肾小管上皮细胞凋亡程度。结果 跟Sham组比较,I/R和metformin+I/R组的BUN、Cr均明显升高;同时,相比I/R组,metformin+I/R组的BUN和Cr均明显降低,且差异均有统计学意义(P<0.05)。免疫组化检测结果显示,跟Sham组比较,I/R组和metformin+I/R组Bcl-2、Bax、caspase-3表达明显增高;跟I/R组比较,metformin+I/R组Bax、caspase-3表达明显降低,同时Bcl-2的表达明显强于I/R组。Tunel结果显示,相比Sham组,I/R和metformin+I/R组肾小管上皮细胞凋亡数目明显增多;同时,metformin+I/R组肾小管上皮细胞凋亡数目明显少于I/R组,且差异有统计学意义(P<0.05)。结论 二甲双胍预处理能改善大鼠肾缺血再灌注损伤过程中肾功能损害,其机制可能与抑制肾小管上皮细胞凋亡有关。
中文关键词:二甲双胍  缺血再灌注  凋亡  肾脏
 
Effects and Mechanisms of Metformin Preconditioning on Renal Ischemia-reperfusion Injury in Rats
Abstract:Objective To investigate the effects of metformin preconditioning on renal ischemia-reperfusion injury in rats and its possible mechanism. Methods A total of 30 rats which were healthy adult male SD rats weighing 220-250g were randomly divided into 3 groups randomly (n=10):sham operation group (Sham group), ischemia reperfusion group (I/R group) and metformin group (Metformin+I/R group). Group metformin+I/R was given intraperitoneal injection of metformin 0.125mg/(kg·d) before the establishment of I/R model, 14days, while Sham group and I/R group were given equal amount of normal saline. After the end of administration, I/R group and metformin+I/R group used the method of removing the right kidney, clamping the left renal artery and vein 45min after the recovery of perfusion to prepare the rat model of renal ischemia and reperfusion. In Sham group, resection of the right kidney was performod, without clamping the left renal artery and vein. Determination of serum creatinine in the blood from inferior vena cava at 24h after reperfusion (Cr) and urea nitrogen (BUN) concentration was performed. Then the rats were sacrificed.Renal tissue morphology, immunohistochemical expression of Bcl-2, Bax and caspase-3 were observed under light microscope. At the same time, Tunel staining were used to evaluate the degree of renal tubular epithelial cell apoptosis. Results Compared with sham group, BUN and Cr in the I/R group and metformin+I/R group were significantly higher than those in the sham group, while the BUN and Cr in the Metformin+I/R group were significantly lower than those in the I/R group, and the difference was statistically significant (P<0.05). Immunohistochemistry showed that, compared with Sham group,in I/R group and metformin+I/R group, the expression of Bcl-2, Bax and caspase-3 increased significantly. Compared with I/R group, in metformin+I/R group, caspase-3 and bax expression decreased significantly, while the expression of Bcl-2 was stronger than I/R group. Tunel results showed that compared with Sham group, I/R metformin+I/R group and the number of apoptosis of renal tubular epithelial cells was significantly increased in metformin+I/R group. At the same time, the apoptosis of renal tubular epithelial cells was significantly less than I/R group, and the difference was statistically significant. Conclusion Metformin pretreatment can improve renal damage in rats with renal ischemia-reperfusion injury, which may be related to the inhibition of renal tubular epithelial cell apoptosis.
keywords:Metformin  Ischemia reperfusion  Apoptosis  Kidney
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