Caspase-1/-11通过剪切GSDMD参与LPS诱导的脓毒症急性肾损伤  

Caspase-1/-11 participates in LPS-induced sepsis-associated acute kidney injury by cleaving GSDMD

作　　者：翟斌 马丽莎 申瑞芹 余剑 陶宜楠 徐爱萍 邵德翠[1] ZHAI Bin;MA Li-Sha;SHEN Rui-Qin;YU Jian;TAO Yi-Nan;XU Ai-Ping;SHAO De-Cui(Cell Electrophysiology Laboratory,Wannan Medical College,Wuhu 241002,China;Human Anatomy Experiment and Training Center,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院细胞电生理研究室,芜湖241002 [2]皖南医学院人体解剖学实验实训中心,芜湖241002

出　　处：《生理学报》2023年第1期10-16,共7页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China (No. 81400695);the Natural Science Foundation of Anhui Province,China (No. 1508085QH153, 1908085QC132);the Natural Science Research Project Fund for Colleges and Universities in Anhui Province,China (No. KJ2019A0408);the Foundation of Wannan Medical College (No. WK202024)。

摘　　要：肾小管上皮细胞死亡是急性肾损伤(acute kidney injury, AKI)的重要原因。本文旨在研究由Gasdermin D (GSDMD)介导的细胞焦亡是否参与脂多糖(lipopolysaccharide, LPS)诱导的脓毒症AKI,并探讨caspase-1、caspase-11焦亡通路在其中的作用。将小鼠分为4组:野生型(WT)组、野生型-LPS (WT-LPS)组、GSDMD基因敲除型(KO)组和GSDMD基因敲除型-LPS (KOLPS)组,通过腹腔注射LPS (40 mg/kg)构建脓毒症AKI模型,取血液样品测定血清肌酐、尿素氮的浓度;取小鼠肾组织标本进行HE染色,观察肾组织病理学变化;用Western blot检测焦亡通路相关蛋白的表达量。结果显示,与WT组相比,WT-LPS组血清肌酐、尿素氮的浓度明显升高(P <0.01);与WT-LPS组相比,KO-LPS组血清肌酐和尿素氮显著降低(P <0.01)。HE染色结果显示,GSDMD敲除后LPS诱导的肾小管扩张得到缓解。和WT组相比,WT-LPS组白介素1β (interleukin-1β, IL-1β)、GSDMD和GSDMD-N蛋白表达水平显著上调,而GSDMD基因敲除显著下调LPS诱导的IL-1β、caspase-11、pro-caspase-1和caspase-1(p22)蛋白表达。上述结果提示,GSDMD介导的细胞焦亡参与LPS诱导的脓毒症AKI,caspase-1和caspase-11可能参与GSDMD的剪切。The present study was aimed to investigate whether Gasdermin D(GSDMD)-mediated pyroptosis participated in lipopolysaccharide(LPS)-induced sepsis-associated acute kidney injury(AKI), and to explore the role of caspase-1 and caspase-11 pyroptosis pathways in this process. The mice were divided into four groups: wild type(WT), WT-LPS, GSDMD knockout(KO) and KO-LPS.The sepsis-associated AKI was induced by intraperitoneal injection of LPS(40 mg/kg). Blood samples were taken to determine the concentration of creatinine and urea nitrogen. The pathological changes of renal tissue were observed via HE staining. Western blot was used to investigate the expression of pyroptosis-associated proteins. The results showed that the concentrations of serum creatinine and urea nitrogen in the WT-LPS group were significantly increased, compared with those in the WT group(P < 0.01);whereas serum creatinine and urea nitrogen in the KO-LPS group were significantly decreased, compared with those in the WT-LPS group(P < 0.01).HE staining results showed that LPS-induced renal tubular dilatation was mitigated in GSDMD KO mice. Western blot results showed that LPS up-regulated the protein expression levels of interleukin-1β(IL-1β), GSDMD and GSDMD-N in WT mice. GSDMD KO significantly down-regulated the protein levels of IL-1β, caspase-11, pro-caspase-1, caspase-1(p22) induced by LPS. These results suggest that GSDMD-mediated pyroptosis is involved in LPS-induced sepsis-associated AKI. Caspase-1 and caspase-11 may be involved in GSDMD cleavage.

关 键 词：急性肾损伤 脂多糖 Gasdermin D 焦亡 

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