补气活血解毒方及依普利酮抑制醛固酮诱导HK2细胞焦亡  

Effect of Buqi Huoxue Jiedu formula and eplerenone on pyroptosis of HK2 cells induced by aldosterone

作　　者：马雪莲 李慧 常奕[2] 王筝[2] 王香婷 王琬瑶[2] 许庆友 MA Xue-lian;LI Hui;CHANG Yi;WANG Zheng;WANG Xiang-ting;Wang Wan-yao;XU Qing-you(Hebei Key Lab of Integrative Medicine on Liver-Kidney Patterns;Hebei University of Chinese Medicine,Shijiazhuang 050091,China)

机构地区：[1]河北省中西医结合肝肾病证研究重点实验室 [2]河北中医学院,河北石家庄050091

出　　处：《中国药理学通报》2021年第12期1770-1775,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81873251);河北省卫计委医学重点课题(No 20180889);河北省省属高校基本科研业务费专项项目(JCYJ2021004);河北中医学院大学生创新创业项目(No 202014432178)。

摘　　要：目的观察补气活血解毒方含药血清对人肾小管上皮细胞(HK2)焦亡作用。方法1μmol·L^(-1)醛固酮诱导HK2细胞焦亡,采用补气活血解毒方含药血清和依普利酮干预。实验分组:对照组(Cont)、醛固酮组(Ald)、补气活血解毒方干预组(TCM),依普利酮干预组(EPL),24 h药物干预后收集细胞检测。TUNEL标记HK2细胞DNA损伤;细胞免疫荧光和Western blot检测盐皮质激素受体NR3C2及其活化下游因子SGK-1、NF-κB表达,检测焦亡通路NLRP3、caspase-1和IL-1β蛋白表达。结果与对照组比较,醛固酮组TUNEL阳性细胞明显增多,NR3C2核转位细胞增多,SGK-1、NF-κB、NLRP3、caspase-1和IL-1β蛋白表达增强。补气活血解毒方血清和依普利酮干预后TUNEL阳性细胞明显下降,NR3C2核转位细胞减少,焦亡相关蛋白表达下调。结论补气活血解毒方含药血清和依普利酮均可抑制醛固酮诱导的HK2细胞焦亡,减轻炎性损伤及肾脏固有细胞损失,减缓梗阻性肾病进展。Aim To study the effect of Buqi Huoxue Jiedu formula(TCM)reducing the pyroptosis of HK2(human proximal tubular epithelial cells)induced by aldosterone(Ald).Methods HK2 cells were induced to pyroptosis by aldosterone(1μmol·L^(-1))and then intervented by medicated serum containing TCM(10%),as well as eplerenone(10μmol·L^(-1)).Control group(Cont group),Ald group,Ald+medicated serum containing TCM group(TCM group),Ald+eplerenone group(EPL group)were divided.After 24 h drug intervention,cells were collected for detection.DNA damage was tested by TUNEL.The protein expression of NR3C2,SGK-1,NF-κB,as well as the signaling pathway of pyroptosis related proteins including NLRP3,caspase-1,IL-1βwere detected by immunofluorescence staining and Western blot.Results Compared with control group,TUNEL positive cells,NR3C2 in nuclei,SGK-1,NF-κB,NLRP3,caspase-1 and IL-1βprotein were all up-regulated in Ald group,which were all significantly reduced after intervented by either medicated serum containing TCM or eplerenone.Conclusions Both medicated serum containing TCM and eplerenone can inhibit HK2 cell pyroptosis induced by aldosterone,thereby alleviating the inflammation and the loss of intrinsic cells in kidney,and so as to delay the progression of obstructive nephropathy.

关 键 词：依普利酮 补气活血解毒方 HK2细胞 焦亡 醛固酮 梗阻性肾病 

分 类 号：R-332[医药卫生] R289.5