Contrast induces kidney epithelial cell apoptosis through NRLP3 inflammasome pathway
10.3760/cma.j.issn.1001-7097.2018.01.007
- VernacularTitle:对比剂激活NLRP3炎症体通路诱导肾小管上皮细胞凋亡
- Author:
Jianxiao SHEN
1
;
Ling WANG
;
Na JIANG
;
Xinghua SHAO
;
Chaojun QI
;
Zhen ZHANG
;
Yijun ZHOU
;
Shu LI
;
Shan MOU
;
Leyi GU
;
Zhaohui NI
Author Information
1. 200240,上海交通大学医学院附属仁济医院肾脏科
- Keywords:
Contrast media;
Apoptosis;
Apoptosis regulatory proteins;
Nod-like receptor protein 3 inflammasome
- From:
Chinese Journal of Nephrology
2018;34(1):36-43
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of pyrin domain 3 (NLRP3) inflammasome in the process of contrast induced human kidney cell apoptosis.Methods Human kidney 2 (HK-2) cells were cultured in DMEM-F12 medium with 5％ FBS.Cells were divided into control group,Contrast group (O group),NLRP3-siRNA+Iohexol group (si-NLRP3+O group),ASC-siRNA+Iohexol group (si-ASC+O group),and mannitol group (M group).Different concentrations of hypotonic contrast agent were added to HK-2 cell culture plates for 24,48 and 72 h.Flow cytometry was used to detect apoptosis.NLRP3 and ASC mRNA expressions were detected by RT-PCR.The expressions of NLRP3,ASC,caspase-8/cleaved caspase-8,Bcl-2/Bax,caspase-1/cleaved caspase-1,and caspase-3/cleaved caspase-3 protein were detected by Western blot.The levels of interleukin (IL) 1β and IL-18 in supernatant were detected by ELISA.Results Compared with the control group,the rate of apoptotic cells,as well as the expressions of NLRP3,ASC and cleaved caspase-1 proteins were increased in HK-2 cells of contrast group.The expressions of NLRP3 and ASC mRNA in the contrast group also increased,so did IL-1β and IL-18 levels (all P＜0.05),suggesting that NLRP3 inflammasome in HK-2 cells was activated by contrast.Compared with the control group,the expressions of cleaved caspase-8,Bax and cleaved caspase-3 protein were increased,and the expression of anti-apoptotic protein Bcl-2 was decreased (all P ＜ 0.05).Compared with the contrast group,the rate of apoptotic cells in the si-NLRP3 + contrast group and si-ASC + contrast group was significantly decreased;the expression of cleaved caspase-1 was decreased;the expressions of Bax and cleaved caspase-3 were decreased,and Bcl-2 level was increased.The expressions of IL-1β and IL-18 in the supernatant of cells were decreased (all P ＜ 0.05).Conclusion Contrast agent can activate the NLRP3 pathway in HK-2 cells and induce apoptosis,which could be reduced by blocking the NLRP3 pathway.
