Piezo 1通过调节肾小管上皮细胞炎症和凋亡参与脓毒症相关急性肾损伤  

Piezo 1 participates in sepsis-associated acute kidney injury by regulating inflammation and apoptosis

作　　者：邓淑婷 袁也 李博厚 董晓颖 何朝生[2] 刘双信 DENG Shuting;YUAN Ye;LI Bohou;DONG Xiaoying;HE Chaosheng;LIU Shuangxin(School of Medicine,South China University of Technology,Guangzhou 510006,China;Department of Nephrology,Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China)

机构地区：[1]华南理工大学医学院,广州510006 [2]南方医科大学附属广东省人民医院(广东省医学科学院)肾内科

出　　处：《肾脏病与透析肾移植杂志》2024年第5期408-416,共9页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金项目(81870508,81873616,82170730);广东省自然科学基金项目(2022A1515012374,2023A1515010024);广东省登峰计划项目(DFJH201901)。

摘　　要：目的:探讨Piezo 1在脓毒症相关急性肾损伤(SA-AKI)的发病机制中的作用。方法:利用脂多糖(LPS)处理C57BL/6小鼠和人近端肾小管上皮细胞(HK-2细胞)构建SA-AKI模型,Western Blot和免疫荧光检测Piezo 1的表达。肽毒素铲形机械毒素4(GsMTx4)干预SA-AKI小鼠,HE和PAS染色检测小鼠肾组织病理改变。用Piezo 1 siRNA或Yoda 1干预LPS诱导的HK-2细胞,用Western Blot、实时荧光定量PCR(RT-qPCR)检测肾小管损伤标志物、炎症因子和凋亡标志蛋白的表达;ELISA法检测细胞上清炎症因子水平;流式细胞术检测细胞凋亡率。Piezo 1 siRNA处理后,用Fluo-4 AM钙离子(Ca^(2+))荧光探针检测细胞内Ca^(2+)含量;Western Blot检测钙蛋白酶2(Calpain 2)、整合素β1(Integrinβ1)的表达。PD151746干预后检测Integrinβ1的表达。Yoda1和PD151746同时干预LPS诱导的细胞,ELISA法检测细胞上清炎症因子水平和流式细胞术检测细胞凋亡率。结果:Piezo 1在SA-AKI小鼠肾小管和LPS处理的HK-2细胞中表达均增加。GsMTx4抑制Piezo 1可减轻SA-AKI小鼠肾小管损伤。沉默Piezo 1可减少LPS诱导的HK-2细胞损伤、炎症和细胞凋亡,然而,Yoda 1进一步加重上述损伤。通过沉默Piezo 1,可减少LPS诱导的Ca^(2+)细胞内流和Calpain 2、Integrinβ1的表达。PD151746抑制Calpain 2可减少Integrinβ1的表达,并抑制Yoda 1引起的HK-2细胞炎症和细胞凋亡。结论:Piezo 1在SA-AKI肾小管上皮细胞中表达上调,可能是通过激活Ca^(2+)/Calpain 2/Integrinβ1通路引起炎症和细胞凋亡,参与SA-AKI的发生发展。Objective:To investigate the role and possible mechanism of Piezo 1 in LPS-induced sepsis-associated acute kidney injury(SA-AKI).Methodology:Establish in vivo and in vitro models of SA-AKI in mice and human proximal renal tubular epithelial cells(HK-2 cells)by LPS,and immunofluorescence and Western blot were used to detect Piezo 1 expression.GsMTx4 was injected into SA-AKI mice,and the renal pathology and renal function were detected.Piezo 1 siRNA or Yoda 1 were used to intervene in LPS-induced HK-2 cells,then the expressions of renal tubular injury markers,inflammatory factors and apoptosis-related marker proteins were detected by Western blot and RT-qPCR,inflammatory factors were detected by ELISA and apoptosis rate was detected by flow cytometry.After treated with Piezo 1 siRNA,the intracellular Ca^(2+)concentration was detected with a Fluo-4 AM calcium ion fluorescent probe and the expressions of Calpain 2 and Integrinβ1 were detected.Integrinβ1 expression was detected after PD151746 intervention.After Yoda1 and PD151746 simultaneously intervened in LPS-induced cells,inflammatory factors were detected by ELISA and apoptosis rate was detected by flow cytometry.Results:Piezo 1 expression was upregulated in the kidneys of mice with SA-AKI and HK-2 cells induced by LPS.GsMTx4 ameliorated renal pathology and renal function damage in SA-AKI mice.Silencing Piezo 1 reduced LPS-induced HK-2 cell damage,inflammation,and apoptosis;however,the above damage was further exacerbated after Yoda 1 intervention.Mechanistically,Piezo 1 knockdown reduced LPS-induced intracellular Ca^(2+)and the expression of Calpain 2 and Integrinβ1.PD151746 reduced the expression of integrinβ1 and inhibited inflammation and apoptosis caused by Yoda 1.Conclusion:Piezo 1 expression is upregulated in renal tubular epithelial cells with SA-AKI,possibly causes inflammation and apoptosis by activating the Ca^(2+)/Calpain 2/Integrinβ1 pathway,and which is involved in the progression of SA-AKI.

关 键 词：Piezo 1 脓毒症相关急性肾损伤 炎症 凋亡 

分 类 号：R459.7[医药卫生—急诊医学] R692[医药卫生—治疗学]