器官移植2025，Vol.16Issue(3)：416-424,9.DOI:10.12464/j.issn.1674-7445.2025072
下调USP46通过抑制NLRP3去泛素化减轻细胞焦亡改善肾小管上皮细胞缺氧/复氧损伤
Downregulation of USP46 alleviates hypoxia/reoxygenation-induced pyroptosis in renal tubular epithelial cells by inhibiting NLRP3 deubiquitination
摘要
Abstract
Objective To investigate the role and mechanism of ubiquitin-specific protease 46(USP46)in hypoxia/reoxygenation(H/R)-induced pyroptosis of renal tubular epithelial cells.Methods Renal tubular epithelial cells were divided into negative control siRNA group(si-CTL group),USP46 knockdown group(si-USP46 group),negative control siRNA+H/R treatment group(si-CTL+H/R group),and USP46 knockdown+H/R treatment group(si-USP46+H/R group).Flow cytometry was used to detect cell apoptosis in each group.Real-time quantitative polymerase chain reaction(RT-qPCR)was employed to measure the messenger RNA(mRNA)expression of USP46,NOD-like receptor protein 3(NLRP3),gasdermin D(GSDMD),interleukin(IL)-18,and IL-1β.Western blotting was used to detect the protein expression of USP46,NLRP3,GSDMD,and cleaved cysteinyl aspartate specific proteinase(C-Caspase)-1.The levels of inflammatory factors and lactate dehydrogenase(LDH)in the cell supernatants were detected,and the levels of reactive oxygen species(ROS)and malondialdehyde(MDA)in the cells were detected.Co-immunoprecipitation was used to verify the interaction between USP46 and NLRP3.Results Compared with the si-CTL group,the si-CTL+H/R group exhibited increased cell apoptosis,elevated protein expression of USP46,NLRP3,GSDMD-N and C-Caspase-1,increased mRNA expression of USP46,NLRP3,GSDMD,IL-18 and IL-1β,higher levels of IL-18,IL-1β,TNF-α and LDH,and increased ROS and MDA levels(all P<0.05).Compared with the si-CTL+H/R group,the si-USP46+H/R group showed decreased cell apoptosis,reduced protein expression of USP46,NLRP3,GSDMD-N and C-Caspase-1,decreased mRNA expression of USP46,GSDMD and IL-18,lower levels of IL-18,IL-1β,TNF-α and LDH,and decreased ROS and MDA levels(all P<0.05).Co-immunoprecipitation results indicated that USP46 could bind to NLRP3.Conclusions Downregulation of USP46 alleviates H/R-induced pyroptosis in renal tubular epithelial cells,possibly by inhibiting USP46-dependent NLRP3 deubiquitination and promoting NLRP3 ubiquitination and degradation.关键词
肾移植/缺血-再灌注损伤/细胞焦亡/泛素化特异性蛋白酶 46/NOD样受体蛋白 3/炎症小体/去泛素化/氧化应激Key words
Kidney transplantation/Ischemia-reperfusion injury/Pyroptosis/Ubiquitin-specific protease 46/NOD-like receptor protein 3/Inflammasome/Deubiquitination/Oxidative stress分类
医药卫生引用本文复制引用
顾世琦,倪海强,彭宣,曹孝天,宫念樵..下调USP46通过抑制NLRP3去泛素化减轻细胞焦亡改善肾小管上皮细胞缺氧/复氧损伤[J].器官移植,2025,16(3):416-424,9.基金项目
国家自然科学基金(82170772、82370759) （82170772、82370759）
