肾损伤分子-1参与碘对比剂致人肾小管上皮细胞体系HK-2凋亡  

Kidney injury molecule-1 is involved in the apoptosis of renal tubular cell line HK-2 induced by iodine contrast

作　　者：于锐[1] 张新茹 王丹丹[1] 何平[1] 白瑜[1] 田密[1] 张蓓茹[1] YU Rui;ZHANG Xin-ru;WANG Dan-dan;HE Ping;BAI Yu;TIAN Mi;ZHANG Bei-ru(Department of Nephrology, Shengjing Hospital of China Medical University, Shenyang 110004;Department of Nephrology, Central Hospital of Sujiatun District, Shenyang 110101, China)

机构地区：[1]中国医科大学附属盛京医院肾内科,辽宁沈阳110004 [2]沈阳市苏家屯区中心医院肾内科,辽宁沈阳110101

出　　处：《基础医学与临床》2020年第9期1206-1211,共6页Basic and Clinical Medicine

基　　金：辽宁省自然基金指导计划(2019-ZD-0771)。

摘　　要：目的探讨肾损伤分子-1(KIM-1)在对比剂所致肾小管上皮细胞凋亡中的作用及其可能机制。方法培养人肾小管上皮细胞系HK-2,给予75 mg/mL的碘海醇(对比剂)处理不同时间,用Western blot检测KIM-1蛋白表达。设计siRNA靶向干扰KIM-1基因表达,将细胞分为对照组(A)、对比剂组(B)、对比剂+空载组(C)、对比剂+KIM-1-siRNA组(D);除对照组,每组给予75 mg/mL碘海醇处理2 h,通过流式细胞计量术以及检测Bax和Bcl-2的表达评估细胞凋亡;观察各组丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)和单核细胞趋化蛋白1(MCP-1)水平;评估KIM-1在对比剂导致HK-2细胞损伤中的可能机制。结果75 mg/mL碘海醇处理HK-2细胞后,KIM-1表达增多,2 h时最明显。当给予碘海醇处理HK-2细胞2 h后,细胞发生明显凋亡,同时ROS和MCP-1水平上升(P<0.05),而SOD和MDA水平下降(P<0.05);转染siRNA靶向干扰KIM-1基因表达后,相比较对比剂组,细胞凋亡程度明显减轻,同时伴随ROS和MCP-1水平下降,SOD和MDA水平上升(P<0.05)。结论碘对比剂可导致HK-2细胞的KIM-1表达增加,而KIM-1可能通过促进氧化应激及炎性反应参与碘对比剂所致HK-2细胞凋亡。Objective To explore the role and the possible mechanism of kidney injury molecule-1(KIM-1)in the contrast-induced apoptosis of renal tubular epithelial cell.Methods Renal tubular epithelial cells line HK-2 was treated with contrast iohexol(75 mg/mL)and the expression of KIM-1 was examined by Western blot.The siRNA sequences were designed to interfere with KIM-1 gene expression.HK-2 cells were divided into four groups:control group(A),contrast group(B),contrast/vehicle group(C)and contrast/KIM-1-siRNA group(D).Each group was treated with 75 mg/mL iohexol for 2 h except control group.Flow cytometry and Bax and Bcl-2 protein expression were used to evaluate cell apoptosis.MDA,ROS,SOD,MDA,and MCP-1 were also examined in each group to evaluate the possible mechanism of KIM-1 involving in renal tubular epithelial cell injury induced by contrast agent.Results The expression of KIM-1 in HK-2 cells increased under iohexol exposure and reached the peak at 2 h.Iohexol treatment induced the apoptosis of HK-2 cells significantly.Meanwhile,the up-regulation of ROS and MCP-1 as well as the down-regulation of SOD and MDA was found(P<0.05).Knockdown of KIM-1 by siRNA relieved the iohexol-induced apoptosis of HK-2 cells and partly dampened the up-regulation of ROS and MCP-1 and the down-regulation of SOD and MDA.Conclusions Iohexol can stimulate the expression of KIM-1 in renal tubular epithelial cells.KIM-1 may function in contrast-induce apoptosis of renal tubular epithelial cells by promoting oxidative stress and inflammatory reaction.

关 键 词：对比剂肾病 肾损伤分子-1 肾小管上皮细胞 凋亡 氧化应激 

分 类 号：R34[医药卫生—基础医学]