摘要
目的:探讨磷脂酶D2抑制剂CAY10594对急性肾损伤(AKI)的保护作用及其潜在机制。方法:通过对脂多糖(LPS)处理后的人源急性单核白血病细胞系(THP-1)进行转录组测序(RNA-seq),筛选差异表达基因,进一步结合连通性图谱(CMap)数据库预测潜在药物分子,筛选出CAY10594作为候选干预药物。在LPS诱导C57BL/6小鼠脓毒症相关AKI及肾脏缺血再灌注损伤(IRI)诱导AKI两种模型中进行验证,并通过免疫荧光、Western Blot和ELISA等手段评估CAY10594的效果。结果:两种AKI模型小鼠均出现炎症因子水平升高、肝肾功能损伤及组织病理肾小管损伤。CAY10594干预显著降低血清炎症因子水平,改善谷丙转氨酶、谷草转氨酶、肌酐及尿素氮水平(P<0.05);组织病理学检查显示干预组肾小管坏死和巨噬细胞浸润显著减轻,并伴随肾脏F4/80阳性巨噬细胞浸润减少及NOD样受体家族pyrin结构域含3型炎症小体(NLRP3)及肾损伤分子1(KIM-1)表达下调。Western Blot结果证实,CAY10594通过抑制核因子κB(NF-κB)通路的激活,降低NLRP3表达及降低KIM-1蛋白水平。结论:CAY10594通过抑制炎症反应及炎细胞浸润改善AKI,其可能机制是通过抑制F4/80-NF-κB-NLRP3轴来减少巨噬细胞介导的炎症反应,具有潜在的临床应用价值。
Objective:To investigate the protective effect of the phospholipase D2 inhibitor CAY10594 on acute kidney injury(AKI)and its underlying mechanisms.Methodology:Transcriptional profiling(RNA-seq)was performed on the human acute monocytic leukemia cell line THP-1 treated with lipopolysaccharide(LPS)to identify differentially expressed genes.Potential drug molecules were predicted by integrating the Connectivity Map(CMap)database,and CAY10594 was selected as a candidate intervention.Two AKI models were established:LPS-induced sepsis-related AKI and renal ischemia-reperfusion injury(IRI)-induced AKI in C57BL/6 mice.The effect of CAY10594 was assessed by immunofluorescence,Western blot,and ELISA.Results:Both the LPS and IRI models showed elevated levels of inflammatory cytokines,liver and kidney dysfunction,and renal tubular damage.Treatment with CAY10594 significantly reduced serum inflammatory cytokine levels and improved liver and kidney function markers(ALT,AST,CREA,and UREA,P<0.05).Histopathological examination showed that CAY10594 intervention significantly alleviated tubular necrosis and macrophage infiltration,accompanied by reduced F4/80+macrophage infiltration and downregulation of NLRP3 and KIM-1 expression in the kidneys.Western blot analysis confirmed that CAY10594 inhibited NF-κB pathway activation,reduced NLRP3 expression,and lowered KIM-1 protein levels.Conclusion:CAY10594 improves acute kidney injury by inhibiting inflammation and inflammatory cell infiltration.Its potential mechanism involves suppression of the F4/80-NF-κB-NLRP3 axis to reduce macrophage-mediated inflammatory responses,suggesting potential clinical applicability.
作者
古晓燕
卢颖辉
南迪
龚德华
GU Xiaoyan;LU Yinghui;NAN Di;GONG Dehua(National Clinical Research Center for Kidney Diseases,Jinling Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing 210016,China)
出处
《肾脏病与透析肾移植杂志》
2025年第6期532-540,共9页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
国家自然科学基金面上项目(81470992)
东部战区总医院管理基金(2023JCYJYB124)。
