激酶锚定蛋白1抑制线粒体分裂减轻肾脏缺血再灌注损伤

Kinase anchor protein 1 alleviates renal ischemia-reperfusion injury by inhibiting mitochondria division

目的:探讨激酶锚定蛋白1(AKAP1)在肾脏缺血再灌注损伤(IRI)中的作用及机制。方法:首先构建小鼠肾脏缺血再灌注模型,夹闭双侧肾动脉缺血28 min,再灌注24 h后采血并留取肾组织,检测血清肌酐、尿素氮、病理损伤及AKAP1表达情况,其次将小鼠肾小管上皮细胞培养至第2天进行干预,缺氧过程(H)将细胞置入1%O_(2)、5%CO_(2)、37℃三气培养箱内无糖无血清依次培养3 h,复氧(R)时更换含10%胎牛血清低糖培养基正常培养6 h。观察对照组和各缺复氧(H/R)组再复氧6 h时肾小管上细胞一般形态,最后通过转染过表达AKAP1的方式检测H/R各组ATP含量,线粒体膜电位以及线粒体Mito-Red染色情况。结果:AKAP1在肾脏缺血再灌注后表达下降(P<0.05);小鼠肾小管上皮细胞在H/R处理后出现AKAP1表达下降以及线粒体功能障碍(P<0.05);AKAP1通过线粒体动力相关蛋白1调控H/R诱导线粒体损伤(P<0.05)。结论:AKAP1可以通过抑制线粒体功能障碍减轻肾脏IRI。

Objective:To investigate the role of A kinase anchor protein 1(AKAP1)in renal ischemia reperfusion injury(IRI)and its mechanism.Methodology:The mouse renal ischemia reperfusion model was established by bilateral renal arteries clamped for 28 min of ischemia.After 24 h of reperfusion,blood samples were collected and kidney tissues were retained to detect serum creatinine,urea nitrogen,pathological injury and AKAP1 expression;Then the mouse proximal tubule cells were cultured to the second day for intervention of hypoxia(H).The cells were placed into a three-gas incubator with 1%O_(2),5%CO_(2),37℃and were cultured without sugar or serum successively for 3h.Then the cells were reoxygenated(R)and replaced with lowglucose DMEM containing 10%fetal bovine serum for normal culture for 6h.The general morphology of renal tubule cells in control group and each H/R group were observed after reoxygenation for 6h.Finally,ATP content,mitochondrial membrane potential and mitochondrial MitoRed staining in each H/R group were detected by transfection with AKAP1 overexpression.Results:AKAP1 expression decreased after renal ischemia reperfusion(P<0.05).AKAP1 expression was decreased and mitochondrial dysfunction was observed in renal tubular epithelial cells after deoxygenation(P<0.05).AKAP1 induced mitochondrial damage through Drp1 regulation of hyporexia(P<0.05).Conclusion:AKAP1 reduce renal ischemiareperfusion injury by inhibiting mitochondrial dysfunction.