Department of Nephrology, Henan Provincial Key Laboratory of Kidney Disease and Immunology, Henan Provincial Clinical Research Center for Kidney Disease, People's Hospital of Zhengzhou University and Henan Provincial People's Hospital, 7 Weiwu Road, Jinshui District, Zhengzhou, Henan 450053, China.
Children's Heart Center, People's Hospital of Zhengzhou University, Zhengzhou, Henan 450053, China.
Ecotoxicol Environ Saf. 2024 Oct 1;284:116944. doi: 10.1016/j.ecoenv.2024.116944. Epub 2024 Aug 28.
Aristolochic acid (AA) exposure is a severe public health concern worldwide. AAs damage the kidney with an inevitable acute phase that is similar to acute kidney injury (AKI). Gasdermin E (GSDME) is abundant in the kidney; thus; it-mediated pyroptosis might be essential in connecting cell death and inflammation and promoting AAs-AKI. However, the role and exact mechanism of pyroptosis in AAs-AKI have not been investigated. In this study, aristolochic acid I (AAI) was used to establish AKI models. The expression and translocation results showed GSDME-mediated pyroptosis in AAI-AKI. Knocking down GSDME attenuated AAI-induced cell death and transcription of proinflammatory cytokines. Mechanistic research inhibiting caspase (casp) 3, casp 8, and casp 9 with specific chemical antagonists demonstrated that GSDME was activated by cleaved casp 3. Furthermore, the kinase activity of upstream receptor-interacting protein kinase 1 (RIPK1) was significantly elevated, and inhibiting RIPK1 with specific inhibitors markedly improved AAI-induced cell damage. In addition, the level of autophagy was obviously increased. Pretreatment with a specific autophagic inhibitor (3-methyladenine) or knockdown of autophagic genes (Atg5 or Atg7) evidently reduced the activity of RIPK1 and downstream apoptosis and pyroptosis, thus attenuating AA-induced cell injury, which suggested that RIPK1 was a novel link conferring autophagic promotion of pyroptosis. These findings reveal GSDME-mediated pyroptosis for the first time in AAI-induced AKI, propose its novel role in the transcription of cytokines, and demonstrate that autophagy promotes pyroptosis via the RIPK1-dependent apoptotic pathway. This study promotes the understanding of the toxic effects and exact mechanisms of AAs. This will contribute to evaluating the environmental risk of AA exposure and might provide potential therapeutic targets for AA-AKI.
马兜铃酸（AA）暴露是一个严重的全球公共卫生问题。AA 会损害肾脏，导致不可避免的急性期，类似于急性肾损伤（AKI）。Gasdermin E（GSDME）在肾脏中含量丰富；因此，它介导的细胞焦亡可能是连接细胞死亡和炎症并促进 AA-AKI 的关键。然而，细胞焦亡在 AA-AKI 中的作用和确切机制尚未得到研究。在这项研究中，使用马兜铃酸 I（AAI）建立 AKI 模型。表达和易位结果表明，GSDME 介导的细胞焦亡参与了 AAI-AKI。敲低 GSDME 可减轻 AAI 诱导的细胞死亡和促炎细胞因子的转录。用特异性半胱天冬酶（casp）3、casp8 和 casp9 化学拮抗剂抑制的机制研究表明，GSDME 被裂解的 casp3 激活。此外，上游受体相互作用蛋白激酶 1（RIPK1）的激酶活性显著升高，用特异性抑制剂抑制 RIPK1 可显著改善 AAI 诱导的细胞损伤。此外，自噬水平明显增加。用特异性自噬抑制剂（3-甲基腺嘌呤）预处理或敲低自噬基因（Atg5 或 Atg7）可明显降低 RIPK1 及其下游凋亡和细胞焦亡的活性，从而减轻 AA 诱导的细胞损伤，这表明 RIPK1 是一种新的连接，赋予自噬促进细胞焦亡的作用。这些发现首次揭示了 GSDME 介导的细胞焦亡在 AAI 诱导的 AKI 中的作用，提出了其在细胞因子转录中的新作用，并证明自噬通过 RIPK1 依赖性凋亡途径促进细胞焦亡。这项研究促进了对 AA 毒性作用和确切机制的理解。这将有助于评估 AA 暴露的环境风险，并可能为 AA-AKI 提供潜在的治疗靶点。
