Ma Kai, Huang Jingwen, Zhang Jin, Tian Yuan, Hu Jing, Ma Linhao, Wang Changnan
Lab of Stress Injury, School of Life Sciences, Shanghai University, Shanghai 200444, China.
General Practice Department, Shanghai Pudong New District Kangqiao Community Health Service Center, Shanghai 201315, China.
Pathogens. 2025 Apr 30;14(5):439. doi: 10.3390/pathogens14050439.
Sepsis is a life-threatening condition that is characterized by systemic inflammation and organ dysfunction, with adrenal dysfunction being a significant complication. This study aimed to investigate the role of necroptosis and hydrogen sulfide (HS) in sepsis-induced adrenal dysfunction.
A cecal ligation and puncture (CLP)-induced sepsis mouse model was employed. Adrenocortical-specific mixed lineage kinase domain-like pseudokinase (MLKL) knockout (MLKL-KO) and cystathioneine β-synthase (CBS) knockout (CBS-KO) mice were generated using Cre-loxP technology and adrenocortical-specific Cre tool mice. In vitro experiments utilized TNFα-stimulated Y1 adrenocortical cells. The treatments included the HS donor NaHS, TNFα inhibitor R-7050, necroptosis inhibitor NSA and CBS inhibitor AOAA. Pathological assessment involved hematoxylin-eosin (H&E) staining and a Western blot analysis of necroptosis markers (the phosphorylation of MLKL (p-MLKL) and phosphorylation of receptor-interacting protein kinases 1 (p-RIPK1)).
Sepsis induced adrenal congestion, elevated TNFα levels, and activated necroptosis (increased p-MLKL/p-RIPK1) in wild-type mice. HS treatment attenuated adrenal damage, reduced TNFα, and suppressed necroptosis. MLKL knockout reduced septic adrenal dysfunction, whereas CBS knockout exacerbated septic adrenal dysfunction. In vitro, TNFα induced Y1 cell necroptosis, which was reversed by HS or NSA. AOAA exacerbated TNFα-induced necroptosis in Y1 cells.
HS inhibits TNFα-mediated necroptosis, thereby preserving adrenal integrity in sepsis. Targeting the TNFα-necroptosis axis and enhancing endogenous HS production may represent novel therapeutic strategies for sepsis-associated adrenal dysfunction.
脓毒症是一种危及生命的病症，其特征为全身炎症和器官功能障碍，肾上腺功能障碍是一项重要并发症。本研究旨在探究坏死性凋亡和硫化氢（HS）在脓毒症诱导的肾上腺功能障碍中的作用。
采用盲肠结扎和穿刺（CLP）诱导的脓毒症小鼠模型。利用Cre-loxP技术和肾上腺皮质特异性Cre工具小鼠构建肾上腺皮质特异性混合谱系激酶结构域样假激酶（MLKL）基因敲除（MLKL-KO）和胱硫醚β-合酶（CBS）基因敲除（CBS-KO）小鼠。体外实验使用肿瘤坏死因子α（TNFα）刺激的Y1肾上腺皮质细胞。治疗措施包括HS供体硫氢化钠（NaHS）、TNFα抑制剂R-7050、坏死性凋亡抑制剂NSA和CBS抑制剂氨基氧乙酸（AOAA）。病理评估包括苏木精-伊红（H&E）染色以及对坏死性凋亡标志物（MLKL的磷酸化（p-MLKL）和受体相互作用蛋白激酶1的磷酸化（p-RIPK1））进行蛋白质印迹分析。
脓毒症在野生型小鼠中诱发肾上腺充血、TNFα水平升高并激活坏死性凋亡（p-MLKL/p-RIPK1增加）。HS治疗减轻了肾上腺损伤、降低了TNFα水平并抑制了坏死性凋亡。MLKL基因敲除减轻了脓毒症诱导的肾上腺功能障碍，而CBS基因敲除则加剧了脓毒症诱导的肾上腺功能障碍。在体外，TNFα诱导Y1细胞发生坏死性凋亡，而HS或NSA可使其逆转。AOAA加剧了TNFα诱导的Y1细胞坏死性凋亡。
HS抑制TNFα介导的坏死性凋亡，从而在脓毒症中维持肾上腺的完整性。靶向TNFα-坏死性凋亡轴并增强内源性HS生成可能是脓毒症相关肾上腺功能障碍的新型治疗策略。
