右美托咪定激活Akt通路对缺氧复氧肾小管上皮细胞凋亡的影响  

Effect of dexmedetomidine activating Akt Pathway on apoptosis of renal tubular epithelial cells after hypoxia reoxygen

作　　者：翁迪贵 林群[2] 袁锦强 黄庆清 WENG Digui;LIN Qun;YUAN Jinqiang;HUANG Qingqing(Department of Anesthesiology,Mindong Hospital,Fujian Medical University,Ningde 355000 China)

机构地区：[1]福建医科大学附属闽东医院麻醉科,宁德市355000 [2]福建医科大学附属第一医院麻醉科

出　　处：《临床麻醉学杂志》2020年第9期899-904,共6页Journal of Clinical Anesthesiology

摘　　要：目的观察右美托咪定对缺氧复氧诱导肾小管上皮细胞凋亡的作用,并探讨其作用机制。方法将人肾小管上皮细胞分五组干预:对照组(C组),常氧环境中培养28 h;缺氧复氧组(HR组),低氧环境中培养24 h后常氧环境中培养4 h;右美托咪定处理组(D组),复制缺氧复氧模型前以右美托咪定处理2 h;Akt阻断剂Uprosertib处理组(U组),复制缺氧复氧模型前以Akt阻断剂Uprosertib处理1 h;右美托咪定复合Akt阻断剂Uprosertib处理组(DU组),复制缺氧复氧模型前先以Akt阻断剂Uprosertib处理1 h,再以右美托咪定处理2 h。采用Ellsa法检测细胞上清中TNF-ɑ、IL-6、IL-1β浓度,MTT法检测细胞存活率,流式细胞仪检测细胞凋亡率,Western blot法检测Bcl-2、Bax、caspase-3、caspase-9、p-Akt蛋白含量。结果与C组比较,HR组、D组、U组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05),细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与HR组比较,D组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显降低(P<0.05),细胞存活率、Bcl-2、p-Akt蛋白含量明显升高(P<0.05);U组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05),细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与D组比较,U组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05),细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与U组比较,DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显降低(P<0.05),细胞存活率、Bcl-2、p-Akt蛋白含量明显升高(P<0.05)。结论右美托咪定可通过激活Akt信号通路,抑制缺氧复氧造成的肾小管上皮细胞凋亡,发挥保护作用。Objective To observe the effect of dexmedetomidine on apoptosis of renal tubular epithelial cells induced by hypoxia and reoxygenation.Methods Human renal tubular epithelial cells were divided into 5 groups for intervention:control group(group C),cultured in normoxia for 28 h;hypoxia reoxygenation group(group HR),cultured in hypoxic environment for 24 h and then cultured in normoxia for 4 h;dexmedetomidine treatment group(group D),treated with dexmedetomidine for 2 h before replicating the hypoxia reoxygenation model,Akt blocker Uprosertib treatment group(group U),blocked with Akt 1 h before the replication of the hypoxia-reoxygenation model Uprosertib treatment 1 h;dexmedetomidine combined with Akt blocker Uprosertib treatment group(group DU),before copying the hypoxic-reoxygenation model,the Akt blocker Uprosertib treatment 1h,then dexmedetomidine treatment 2 h.Ellsa method was used to detect the concentration of TNF-ɑ,IL-1βand IL-6 in cell supernatant,MTT method was used to detect cell survival rate,flow cytometry was used to detect cell apoptosis rate,Western blot was used to detect Bcl-2,Bax,caspase 3.Protein content of caspase-9 and p-Akt.Results Compared with group C,the concentrations of TNF-α,IL-6,IL-1β,the apoptosis rate,the protein contents of Bax,caspase-3 and caspase-9 were significantly increased(P<0.05),the cell survival rate,the protein contents of Bcl-2 and p-Akt were significantly decreased in groups HR,D,U and DU(P<0.05).Compared with group HR,the concentrations of TNF-α,IL-6,IL-1β,the apoptosis rate,the protein contents of Bax,caspase-3 and caspase-9 were significantly decreased(P<0.05),the cell survival rate,the protein contents of Bcl-2 and p-Akt were significantly increased in group D and DU(P<0.05),the concentrations of TNF-α,IL-6,IL-1β,the apoptosis rate,the protein contents of Bax,caspase-3 and caspase-9 were significantly increased(P<0.05),the cell survival rate,the protein contents of Bcl-2 and p-Akt were significantly decreased in group U(P<0.05).Compared with group D,the conc

关 键 词：右美托咪定 缺氧复氧 肾小管上皮细胞 AKT通路 凋亡 

分 类 号：R614[医药卫生—麻醉学]