巨噬细胞胆碱能通路对急性缺糖缺氧性肾小管细胞损伤的影响  

The cholinergic pathway alleviates acute oxygen and glucose deprivation induced renal tubular cell injury by reducing the secretion of inflammatory medium of macrophages

作　　者：吴明[1] 吴乐锋 陆俊福 李明利[2] 李赟[3] 徐迹 刘文兰[3] 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518035 [2]深圳市第二人民医院介入治疗科,广东深圳518035 [3]深圳市第二人民医院中心实验室,广东深圳518035 [4]南昌大学第一附属医院重症医学科,南昌330006

出　　处：《解放军医学杂志》2017年第8期663-667,共5页Medical Journal of Chinese People's Liberation Army

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;JCY20150330102401099;JCYJ20160425103130218);深圳市第二人民医院临床-基础一对一桥梁项目基金(2015-16)~~

摘　　要：目的探讨胆碱能通路对急性缺糖缺氧性肾小管细胞损伤的影响。方法分离培养大鼠肾内巨噬细胞,构建巨噬细胞与肾小管上皮细胞共培养体系及缺糖缺氧(OGD)细胞模型,据处理不同将细胞分为OGD组、乙酰胆碱(ACh 100μmol/L)+OGD组和ACh+加兰他敏(Gal 10μmol/L)+OGD组。采用ELISA法检测各组上清液肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)和IL-10的表达;MTT法检测肾小管细胞活力;RT-qPCR及Western blotting检测胆碱酯酶(AChE)m RNA和蛋白表达;比色法检测AChE活性。结果 ACh+OGD组TNF-α和IL-1β水平均低于OGD组,加入Gal之后,TNF-α和IL-1β水平进一步下降;ACh+OGD组肾小管活力高于OGD组,加入Gal之后,肾小管活力进一步增强,差异有统计学意义(P<0.05或P<0.01)。3组之间巨噬细胞ACh E m RNA和蛋白表达差异均无统计学意义(P>0.05)。与OGD组比较,ACh+OGD组与ACh+Gal+OGD组肾小管细胞活力减弱,但ACh+OGD组与ACh+Gal+OGD组间差异无统计学意义(P=0.368)。结论 ACh和Gal可抑制肾脏巨噬细胞分泌炎性介质并抑制胆碱酯酶活性,减轻急性缺氧性肾小管细胞损伤。调控巨噬细胞的胆碱能通路可能是未来急性缺氧性肾损伤的治疗方向。Objective To investigate the effects of cholinergic pathway on acute renal tubular cell injury induced by acute oxygen and glucose deprivation. Methods Rat kidney macrophages were isolated and cultured for constructing macrophages and renal epithelial cells co-cultivating model of oxygen-glucose deprivation （OGD）, and the model cells were divided into three groups： OGD alone group, acetylcholine （ACh 100μmol/L）＋OGD group and ACh ＋ galantamine （Gal 10μmol/L）＋OGD group. The cells underwent OGD treatment for 1 hour, and normally cultured for 24 hours. The expressions of TNF alpha, IL-1 beta, and IL-10 in supernatant fluid were detected by ELISA, the renal tubular cell viability was determined by MTT assay, the expression of acetylcholine esterase （AChE） mRNA and protein were determined by RT-qPCR and Western blotting. The activity of AChE was determined by colorimetric method. Results The expressions of TNF alpha （pg/ml） in OGD, Ach＋OGD group, Ach＋Gal＋OGD groups were 140.2±44.81, 119.46±4.42 and 103.31±1.62 respectively （P〈0.05）, those of IL-1β （pg/ml） were 172.26±13.51,144.34±5.53 and 119.37±11.42 respectively （P〈0.05）, and those of IL-10 （pg/ml） were 181.47±16.01, 173.62±10.12 and 188.36±8.73 respectively （P〉0.05）; The values of renal tubular cell proliferation were 55.02%±6.28%, 66.65%±6.47%, and 79.75%±4.22% respectively （P〈0.01）; the expressions of AChE mRNA in macrophages were 4.07±0.03, 4.22±0.15 and 3.98±0.29 respectively in the three groups （P〉0.05）; those of AchE protein were 0.66±0.07, 0.74±0.04 and 0.67±0.06 respectively （P〉0.05）; The activity of AChE （kU/L） was 0.51±0.02, 0.35±0.05 and 0.32±0.04 respectively （P=0.001, 0.001 and 0.368）. Conclusions ACh and Gal could inhibit the secretion of inflammatory mediators and cholinesterase activity and can reduce the acute hypoxic renal tubular cell injury. The modulation of the cholinergic pathway in macrophages may be the important treatment method for

关 键 词：胆碱能药 炎症介导素类 肾小管 

分 类 号：R692[医药卫生—泌尿科学]