摘要
目的探究高浓度浅蓝霉素A(Cae A)对人肾小管上皮细胞HK2的影响,并探讨NOD样受体热蛋白结构域相关蛋白3(NLRP3)在此过程中发挥的作用。方法采用MTT法测定不同浓度的Cae A对HK2细胞活力的影响;采用实时定量PCR,Western blotting以及免疫荧光法检测高浓度Cae A对肾损伤分子1(KIM-1)以及NLRP3表达情况的影响;同时用实时定量PCR方法,检测高浓度Cae A对白细胞介素-1β(IL-1β)、白细胞介素18(IL-18)、白细胞介素33(IL-33)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子α(TNF-α)的mRNA表达情况;随后将HK2细胞分为对照组、高浓度Cae A组和高浓度Cae A加NLRP3抑制剂CY-09组,Western blotting检测KIM-1和NLRP3蛋白的表达变化。结果MTT的结果显示,高浓度Cae A可以抑制HK2细胞活力;实时定量PCR,Western blotting和免疫荧光实验表明,高浓度Cae A可以上调KIM-1和NLRP3的水平,以及IL-1β、IL-18、IL-33、MCP-1、TNF-α的mRNA表达水平,而CY-09能显著下调NLRP3和KIM-1的蛋白水平。结论高浓度的Cae A能明显抑制HK2细胞的活力,诱导HK2细胞发生损伤及炎症反应,有显著的肾毒性,而且该肾毒性是通过激活NLRP3通路来实现的。
Objective To investigate the effect of high concentration of Caerulomycin A(Cae A)on HK2 in renal tubular epithelial cells and to explore the role of cytoplasmic nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)in this process.Methods The effect of different concentrations of Cae A on the viability of HK2 cells was determined by MTT;the expression of kidney injury molecule(KIM-1)and NLRP3 was detected by real-time quantitative PCR,Western blot and immunofluorescence,while the effect of Cae A on the mRNA expression of IL-1β,IL-18,IL-33,MCP-1,TNF-αwas also measured by real-time quantitative PCR.HK2 cells were divided into control group,high concentration of Cae A group and high concentration of Cae A plus NLRP3 inhibitor CY-09 group,and the expression of KIM-1 and NLRP3 protein was detected by Western blot.Results The results of MTT showed that high concentration of Cae A could inhibit HK2 cell viability.Real-time quantitative PCR,Western blot and immunofluorescence assays showed that high concentration of Cae A upregulated the expression of KIM-1 and NLRP3,as well as the mRNA levels of IL-1β,IL-18,IL-33,MCP-1,TNF-α,while CY-09 could down-regulate the expression of NLRP3 and KIM-1.Conclusion High concentration of Cae A significantly inhibited the viability of HK2 cells and induced damage and inflammatory response to HK2 with some nephrotoxicity that might be achieved via NLRP3 pathway.
作者
单润润
庾聚涛
谢嫚嫚
罗秀峰
谢春亚
崔宗宇
刘晓颖
陈奇
Shan Runrun;Yu Jutao;Xie Manman;Luo Xiufeng;Xie Chunya;Cui Zongyu;Liu Xiaoying;Chen Qi(College of Life Sciences,Anhui Medical University,Hefei 230032;School of Pharmacy,Anhui Medical University,Hefei 230032;Second Clinical Medical College,Anhui Medical University,Hefei 230032)
出处
《安徽医科大学学报》
CAS
北大核心
2023年第9期1429-1434,共6页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:41806158)
安徽医科大学校科研基金(编号:2022xkj008)。
关键词
浅蓝霉素A
细胞损伤
细胞炎症
药物肾毒性
caerulomycin A
cellular damage
cellular inflammation
drug nephrotoxicity