碘普罗胺对HK-2细胞ROS-NLRP3炎症小体信号的作用研究  

Iopromide activates ROS-NLRP3 inflammasome signaling in HK-2 cells

作　　者：陈燕玲[1] 罗婷 高昕乐 李海洲 凡栋 赵晓朵 吴秀香[1] 庄晓东[3] 靳俊峰[1] CHEN Yan-ling;LUO Ting;GAO Xin-yue;LI Hai-zhou;FAN Dong;ZHAO Xiao-duo;WU Xiu-xiang;ZHUANG Xiao-dong;JIN Jun-feng(Department of Pathophysiology,Zhuhai Campus of Zunyi Medical University,Zhuhai 519041,Guangdong,China;Zhongshan School of Medical,Sun yat-sen University,Guangzhou 510080,Guangdong,China;Department of Cardiovasology,The First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,Guangdong,China)

机构地区：[1]遵义医学院珠海校区基础教学部,珠海理学博士519041 [2]中山大学北校区中山医学院,广州510080 [3]中山大学附属第一医院心血管内科,广州510080

出　　处：《医学研究生学报》2018年第11期1131-1136,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81560133);贵州省科技合作计划项目(黔科合LH字[2015]7529号)

摘　　要：目的碘普罗胺可引起近端肾小管上皮细胞(HK-2)损伤,但具体机制尚不清楚。文中旨在观察碘普罗胺对HK-2细胞ROS-NLRP3炎症小体信号的作用,探讨碘普罗胺引起HK-2细胞损伤的可能机制。方法实验共分为6组:对照组及碘普罗胺37、74、111、148、185 mgI/mL组。HK-2细胞加入碘普罗胺24 h后,应用2',7'-二氯荧光黄双乙酸盐染色法及流式细胞仪检测细胞内活性氧水平,Western blot法检测NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体相关蛋白(NLRP3、ASC、caspase-1)、IL-1β、TNF-β及NF-κB蛋白表达水平。结果与对照组活性氧水平[2 551.71±84.00]比较,碘普罗胺37、74、111、148、185 mgI/mL组[4103.89±98.89; 4450.12±108.90; 5050.85±606.76; 6210.57±145.74; 7105.13±426.63]均明显升高(P<0.05); Western blot结果显示,HK-2细胞经碘普罗胺处理后NLRP3、ASC、caspase-1、IL-1β、TNF-α蛋白表达水平均较对照组明显增高,且111 mgI/mL及以上浓度的碘普罗胺处理后NF-κB蛋白表达水平也较对照组明显增高(P<0.05)。结论碘普罗胺对HK-2细胞的损伤作用可能与激活ROS-NLRP3炎症小体信号通路有关。Objective Iopromide can induce injury to HK-2 cells, but its exact mechanism remains poorly understood. This study aimed to explore the influence of iopromide on ROS-NLRP3 inflammasome signaling in HK-2 cells. Methods HK-2 human renal tubular epithelial cells were divided into six groups： control and iopromide at 37, 74, 111, 148 and 185 mgI/mL. The HK-2 cells in the latter five groups were treated with different concentrations of iopromide for 24 hours. Then the ROS level in the cells was detected by 2′,7′-Dichlorodihydrofluorescein diacetate staining and flow cytometry and the protein expressions of NLRP3, ASC, caspase-1, IL-1β, NF-κB and TNF-α determined by Western blot. Results The ROS level was significantly increased in the HK-2 cells treated with iopromide at 37 mgI/ml （4 103.89±98.89）, 74 mgI/mL （4 450.12± 108.90 ）, 111 mgI/mL （5 050.85±606.76）, 148 mgI/mL （ 6 210.57± 145.74） and 185 mgI/ml （7 105.13±426.63） as compared with that in the control group （2 551.71±84.00） （ P 〈0.05）. Western blot showed markedly upregulated expressions of NLRP3, ASC, caspase-1, IL-1β and TNF-α in the HK-2 cells in all the latter five groups in comparison with the control （ P 〈0.05） and an increased level of NF-κB after treated with iopromide at ≥111 mgI/ml （ P 〈0.05）. Conclusion Iopromide may induce injury to HK-2 cells by activating the ROS-NLRP3 inflammasome signaling pathway.

关 键 词：碘普罗胺 HK-2细胞 活性氧 NLRP3炎症小体 

分 类 号：R691.6[医药卫生—泌尿科学]