急性缺糖缺氧通过增强胆碱酯酶表达促进肾小管细胞的炎性损伤  

Acute oxygen and glucose deprivation promotes inflammatory injury of renal tubular cells by enhancing the expression of cholinesterase

作　　者：吴明[1] 吴乐锋 李明利[2] 陆俊福 赖凯[1] 徐迹 刘芬[4] 冯永文[1] 

机构地区：[1]深圳市第二人民医院重症医学科,广东深圳5108035 [2]深圳市第二人民医院介入治疗科,广东深圳5108035 [3]深圳市第二人民医院中心实验室,广东深圳5108035 [4]南昌大学第一附属医院重症医学科,江西南昌330006

出　　处：《实验与检验医学》2016年第4期412-415,434,共5页Experimental and Laboratory Medicine

基　　金：国家自然科学基金(81101410);广东省医学科学技术研究基金(A2016353);广东省深圳市科技创新委项目(JCYJ20130401112313541;JCYJ20150330102401099);深圳市第二人民医院基础-临床桥梁项目(2015)

摘　　要：目的探讨急性缺糖缺氧导致肾小管细胞损伤的机制。方法分离培养大鼠肾内巨噬细胞、肾小管上皮细胞,构建两者共培养(transwell)模型,细胞分成对照组及缺糖缺氧(Oxygen and glucose deprivation,OGD)组,给予缺糖缺氧处理细胞1h后再正常培养24h,ELISA法检测两组上清液TNF-α,IL-1β和IL-10的浓度,噻唑蓝(MTT)检测肾小管细胞活力及RTq PCR及Western Blot检测AChE的mRNA和蛋白的表达。结果在共培养上清液中,对照组与OGD相比,TNF-α(pg/ml):(231.67±36.28)VS(428.67±43.16)(P<0.05),IL-1β(pg/ml):(116.67±21.64)VS(219.63±43.86)(P<0.05),IL-10(pg/ml):(235.67±39.35)VS(432.67±49.72)(P<0.01)。肾小管细胞活力明显降低,分别为(88.41±18.25)VS(46.98±13.87)(P<0.01),OGD组巨噬细胞AChE mRNA和蛋白水平均高于对照组,分别上调了(3.82±0.73)和(2.17±0.46)倍(P<0.01)。结论急性缺糖缺氧增强了肾脏巨噬细胞胆碱酯酶的表达,通过炎症介质,介导了急性缺糖缺氧性肾小管细胞损伤。Objective To investigate the injury mechanism of renal tubular cells induced by acute oxygen and glucose deprivation. Methods Isolation and culture of rat kidney macrophages and renal epithelial cells,constructing co-cultivating model of lacking Oxygen and sugar（Oxygen and glucose deprivation,OGD）,Cells were devided into control group and OGD group,and were given OGD treatment for 1 hour,and then carried out normal culture for up to 24 hours in each group. the expression of TNF alpha,IL-1 beta,IL-10 in supernatant fluid was detected by ELISA,the viability of renal tubular cells was determined by MTT,the expression of mRNA and protein of acetylcholine esterase（AChE） were determined by RT-qPCR and Western Blot respectively.Results The levels of TNF alpha（pg/ml） in the supernatant fluid in cultivation system were（231.67±36.28） in control group VS（428.67±43.16）（P〈0.05） in OGD group,the levels of IL-1β（pg/ml） were（116.67±21.64） in control group VS（219.63±43.86） in OGD group（P〈0.05）,the levels of IL-10（pg/ml） were（235.67±39.35） in control group VS（432.67±49.72） in OGD group（P〈0.01）. The viability of renal tubular cells was（88.41±18.25） VS（46.98±13.87）（P〈0.01）;The levels of mRNA and protein of AChE in OGD group were higher than those in control group,they were raised（3.82±0.73） and（2.17±0.46） times respectively（P〈0.01）.Conclusion Acute oxygen and glucose deprivation enhances the expression of cholinesterase in renal macrophages,the acute injury of renal tubular cells induced by OGD was mediated through inflammatory mediators.

关 键 词：缺糖缺氧 胆碱酯酶 炎症介质 肾小管细胞活力 

分 类 号：R692.6[医药卫生—泌尿科学] R-332[医药卫生—外科学]