目的 基于肠道菌群和蛋白组学探究青蒿琥酯改善脓毒症相关急性肾损伤（SA-AKI）的作用机制。方法 采用随机数字表法将小鼠分为5组：假手术组、模型组和青蒿琥酯低、中、高剂量（75、150、300 mg·kg^-1）组，ig给药5 d后，除假手术组外，实施盲肠结扎穿刺术（Clp）建立SA-AKI模型，假手术组不穿刺和挤出粪便，术后3 d内继续给药。计算肾脏指数，使用生化分析仪检测各组小鼠血清尿素氮（BUN）和肌酐（SCr）水平；苏木素-伊红（HE）染色检测肾脏病理变化；透射电子显微镜观察肾皮质变化；蛋白质组学检测肾脏差异蛋白； 16S rDNA测序分析检测肠道微生物变化；以相关性热图分析宿主蛋白、肠道菌群、肾功能指标及脏器指数的关联性。结果 青蒿琥酯以剂量相关方式增加小鼠体质量，降低肾脏指数，降低SCr和BUN水平（P＜0.05、0.01、0.001），并改善肾小球萎缩、肾脏炎症、线粒体皱缩等病理损伤。蛋白质组学结果显示，青蒿琥酯高剂量组KIAA2013、蛋白磷酸酶4调节亚基3B（PPP4R3B）等显著上调； RNA聚合酶II、I和III亚基L（POLR2L）、DNA拓扑异构酶IIα（TOP2A）、Apaf-1相互作用蛋白（APIP）等显著下调（P＜0.05、0.01、0.001） ，基因本体（GO）及京都基因与基因组百科全书（KEGG）分析表明，青蒿琥酯广泛影响细胞的基础代谢、亚细胞结构定位及分子间相互作用网络，可能通过干预低氧适应、能量代谢、血管生成及脂质代谢等关键生物学过程，从而在SA-AKI中发挥保护作用。16S rRNA测序分析显示，与模型组比较，青蒿琥酯高剂量组的肠道菌群多样性显著增加（P＜0.05）；厚壁菌门与变形菌门丰度显著降低，拟杆菌门丰度回升（P＜0.05）；嗜韧带乳杆菌属丰度明显下降（P＜0.05），鼠杆菌属上升（P＜0.05）。相关性分析显示，TOP2A、POLR2L、APIP等蛋白与特定肠道菌群之间存在相关性； TOP2A、POLR2L和APIP的高表达与体质量下降相关，且与肾脏质量及肾功能损害指标升高相关； g_Enterococcus与g_Rikenella的富集可能与肾脏代偿性增大、肾脏指数升高及肾功能损伤加重密切相关；而g_RF39_unclassified、g_Muribaculaceae_unclassified、g_Odoribacter等菌群丰度升高则可能与肾脏结构完整、肾功能指标改善及肠道微生态稳态维持相关。结论 青蒿琥酯可能通过调节“肠-肾轴”恢复肠道微生态平衡，同时调控肾脏关键蛋白及相关通路，从而减轻SA-AKI。

Objective To investigate the mechanism by which artemisinin derivative artesunate ameliorates sepsis-associated acute kidney injury (SA-AKI) via gut microbiota and proteomics. Methods Mice were randomly divided into five groups using a random number table: Sham group, model group, and low-, medium-, and high-dose artesunate groups (75, 150, and 300 mg·kg^-1). After oral gavage for 5 d, all groups except the sham group underwent cecal ligation and puncture (CLP) to induce SA-AKI; The sham group received only a sham operation without puncture or fecal extrusion. Drug administration continued for 3 days post-surgery. Serum blood urea nitrogen (BUN) and creatinine (SCr) levels were measured using a biochemical analyzer; Hematoxylin-eosin (HE) staining assessed renal histopathological changes; Transmission electron microscopy examined cortical renal alterations; Proteomics identified differentially expressed proteins in the kidneys; 16S rDNA sequencing analyzed gut microbiota composition; And correlation heatmaps evaluated associations among host proteins, gut microbiota, kidney function indicators, and organ indices. Results Artesunate increased body weight in a dose-dependent manner, reduced kidney index, and lowered serum creatinine and BUN levels (P < 0.05, 0.01, 0.001), while ameliorating pathological injuries such as glomerular atrophy, renal inflammation, and mitochondrial shrinkage. Proteomic analysis revealed that KIAA2013 and protein phosphatase 4 regulatory subunit 3B (PPP4R3B) were significantly upregulated in the high-dose artesunate group, whereas RNA polymerase II, I, and III subunit L (POLR2L), DNA topoisomerase IIα (TOP2A), and Apaf-1 interacting protein (APIP) were significantly downregulated (P < 0.05, 0.01, 0.001). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that artesunate broadly influenced cellular basal metabolism, subcellular localization, and molecular interaction networks, potentially exerting protective effects in SA-AKI through modulation of key biological processes including hypoxia adaptation, energy metabolism, angiogenesis, and lipid metabolism. 16S rRNA sequencing showed that compared with the model group, the high-dose artesunate group exhibited significantly enhanced gut microbial diversity (P < 0.05); Abundances of Firmicutes and Proteobacteria decreased markedly, while Bacteroidetes abundance recovered (P < 0.05); Lactobacillus reuteri abundance declined significantly (P < 0.05), whereas Muribaculaceae abundance increased (P < 0.05). Correlation analysis revealed associations between TOP2A, POLR2L, and APIP proteins and specific gut microbes. High expression of TOP2A, POLR2L, and APIP correlated with reduced body weights, and was associated with elevated kidney weight and impaired renal function markers. Enrichment of g_Enterococcus and g_Rikenella may be closely related to compensatory kidney enlargement, increased kidney index, and aggravated renal injury; conversely, increased abundances of g_RF39_unclassified, g_Muribaculaceae_unclassified, and g_Odoribacter were likely linked to preserved renal structure, improved renal function, and maintenance of intestinal microecological homeostasis. Conclusion Artesunate may alleviate SA-AKI by modulating the “gut-kidney axis” to restore intestinal microecological balance and regulate critical renal proteins and their associated pathways.

R965

河南省自然科学基金资助项目（262300422203）；河南省医学科技攻关联合共建项目（LHGJ20250569）；洛阳市科学技术协会河洛青年人才托举工程（2025HLTJ44）