N-乙酰半胱氨酸对醋酸铅致肾小管上皮细胞损伤的保护作用  

Protective effects of N-acetylcysteine on renal tubule epithelial cell injury induced by lead acetate

作　　者：周蓉[1] 徐雅虹[1] 陈闽东[1] 沈杰[1] 冯雪芳[1] 陈希[1] 

机构地区：[1]同济大学附属杨浦医院肾内科,上海市杨浦区中心医院肾内科,上海200090

出　　处：《中国中西医结合急救杂志》2016年第1期52-56,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：上海市卫生和计划生育委员会面上项目

摘　　要：目的观察N-乙酰半胱氨酸（NAC）对醋酸铅致人肾小管上皮细胞系（HK-2）损伤过程中中性粒细胞明胶酶相关脂质运载蛋白（NGAL）表达的影响，并探讨其机制。方法将体外培养至少2～5代对数生长期的HK-2细胞分为空白对照组；醋酸铅刺激组（分别用5、50umol／L刺激以确定最佳刺激浓度）；NAC预处理组（用5p．mol／LNAC预处理2h后加入50umol／L醋酸铅）。采用CCK-8法检i贝4HK-2的增殖能力，用流式细胞术检测HK-2的细胞周期，用酶联免疫吸附试验（ELISA）检测细胞上清液中NGAL水平，用流式细胞术检测细胞活性氧（ROS）含量。结果各浓度醋酸铅刺激后6、12、24、48hHK-2的增殖能力[吸光度（A）值]均较空白对照组显著降低，且以50umol／L醋酸铅刺激组降低更显著（6h为0．53±0．02比0．64±0．02，12h为0．46±0．02比0．65±0．02，24h为0．35±0．02比0．68±0．02，48h为0．26±0．02比0．74±0．02，均P〈0．05），12和24hHK-2的细胞周期均呈现G0／1期阻滞[5umol／L醋酸铅刺激组12h为（56．2±1.2）％，24h为（60．6±0．7）％；50umol／L醋酸铅刺激组12h为（71．2±2．0）％，24h为（79．6±2．8）％]；刺激12、24、48、72h各浓度醋酸铅组HK-2上清液中NGAL含量均较空白对照组明显增加，以50umol／L醋酸铅刺激组刺激72h增加更显著（州L：1．212±0．015比0．749±0．040，P〈0．05）。50umol／L醋酸铅刺激组和NAC预处理组HK-2内ROS含量均较空白对照组显著增多．但NAC预处理组低于醋酸铅刺激组[A值（×10。）：34．72±1．67比68．31±1．53]；50tamol／L醋酸铅刺激组和NAC预处理组刺激24、48hHK-2的增殖能力均低于空白对照组，但NAC预处理组高于50umol／L醋酸铅刺激组，以刺激48h增加更显著（A值：0．65±0．01比0．57±0．01）；50umol／L醋酸铅刺激组和NAC预处理组HK-2上清液中NGAL水平均较空白对照组显著增加，但NAC预处理�Objective To investigate the effect of N-acetylcysteine （NAC） on the expression of neutrophil gelatinase-associated lipocalin （NGAL） in the course of the human renal tubule epithelial cell line （HK-2） injury induced by lead acetate and to approach its possible mechanism. Methods The HK-2 cells cultured in vitro were divided into blank control group, 5 μmol/L and 50μmol/L lead acetate groups, NAC pretreatment group （after pretreatment with 5μmol/L NAC for 2 hours, 50μmol/L lead acetate was added）. The proliferation of HK-2 cells was tested by CCK-8 method, the cell cycle was tested by flow eytometry, and the levels of cell supematant NGAL were tested by enzyme linked immunosorbent assay （ELISA）. The reactive oxygen species （ROS） generation was test with flow cytometry. Results The proliferation of HK-2 cells [absorbance （A） value] was significantly lowered after 6, 12, 24, 48 hours cultivation with each lead acetate compared with those in the blank control group, and the degree of decrease in 50 μmol/L lead acetate group was more obvious （6 hours： 0.53 ±0.02 vs. 0.64±0.02, 12 hours： 0.46 ±0.02 vs. 0.65 ± 0.02, 24 hours： 0.35 ± 0.02 vs. 0.68 ± 0.02, 48 hours： 0.26 ± 0.02 vs. 0.74 ±0.02, all P 〈 0.05）. The cell cycles of HK-2 cells were blocked in GO/1 phase after 12 hours and 24 hours cultivation with 5 and 50μmol/L lead acetate solution [5μmol/L lead acetate group 12 hours： （56.2 ± 1.2）%, 24 hours： （60.6 ± 0.7）%, 50 μmol/L lead acetate group 12 hours： （71.2 ± 2.0）%, 24 hours： （79.6 ± 2.8）%]. After 12,24, 48, 72 hours cultivation with various lead acetate groups, the levels of supernatant NGAL were significantly higher than that in blank control group, and the degree of decrease in 50μmol/L lead acetate group after 72 hours of stimulation was the most significant （1.212 ± 0.015 vs. 0.749 ± 0.040, P 〈 0.05）. In 50 p.mol/L lead acetate group and NAC pretreatment group, the levels of ROS were higher significantly than that i

关 键 词：醋酸铅 中性粒细胞明胶酶相关脂质运载蛋白 肾小管上皮细胞 N-乙酰半胱氨酸 

分 类 号：R692[医药卫生—泌尿科学]