Zhang Nieke, Huang Zhicong, Xia Yi, Tao Shuchun, Wu Tiange, Sun Si, Zhu Yongkun, Jiang Guiya, Lu Xun, Gao Yue, Guo Fangfang, Cao Rui, Chen Shuqiu, Zhang Lei, Zou Xiangyu, Chen Ming, Zhang Guangyuan
Department of Urology, Zhongda Hospital, Southeast University, No. 87 Dingjiaqiao, Hunan Road, Gulou District, Nanjing, 210009, China.
Institute of Urology, Medical School, Southeast University, No. 87 Dingjiaqiao, Hunan Road, Gulou District, Nanjing, 210009, China.
J Nanobiotechnology. 2025 Jun 6;23(1):422. doi: 10.1186/s12951-025-03505-9.
Remote ischemic preconditioning (rIPC) has been reported to protect against kidney ischemia-reperfusion injury (IRI) through the delivery of extracellular vesicles (EVs). Among these, apoptosis-induced compensatory proliferation signaling-related vesicles (ACPSVs) can transmit proliferation signals to surrounding cells. However, the underlying mechanisms remain unclear. This study aimed to investigate the role of ACPSVs in renal IRI following rIPC and to elucidate the associated mechanisms.
We demonstrated that rIPC plasma or ACPSVs alleviated renal damage and inflammation, with the protective effects abolished upon the removal of ACPSVs from the plasma. EVs isolated via differential centrifugation exhibited defining characteristics of ACPSVs. Co-culture experiments revealed that ACPSVs reduced apoptosis and enhanced the viability of HK-2 cells under hypoxia/reoxygenation (H/R) conditions. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses highlighted the critical role of macrophage migration inhibitory factor (MIF) protein in ACPSVs. Using CRISPR/Cas9 technology, we generated MIF-knockout HeLa cells to induce the production of MIF-deficient ACPSVs. The protective effects of ACPSVs were significantly attenuated when MIF was knocked out. Transcriptome sequencing and chromatin immunoprecipitation (ChIP) assays revealed that MIF suppresses dual-specificity phosphatase 6 (DUSP6) expression by promoting H3K9 trimethylation (H3K9me3) in the DUSP6 promoter region, thereby activating the JNK signaling pathway. In rescue experiments, treatment with the DUSP6 inhibitor BCI effectively restored the protective function of MIF-deficient ACPSVs.
This study underscores the protective role of ACPSVs derived from rIPC-treated rats and serum-starved cells against renal IRI through the MIF/DUSP6/JNK signaling axis, offering a potential clinical therapeutic strategy for acute kidney injury induced by IRI.
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The online version contains supplementary material available at 10.1186/s12951-025-03505-9.
据报道，远程缺血预处理（rIPC）通过释放细胞外囊泡（EVs）来保护肾脏免受缺血再灌注损伤（IRI）。其中，凋亡诱导的代偿性增殖信号相关囊泡（ACPSVs）可以将增殖信号传递给周围细胞。然而，其潜在机制仍不清楚。本研究旨在探讨ACPSVs在rIPC后肾IRI中的作用，并阐明相关机制。
我们证明，rIPC血浆或ACPSVs可减轻肾脏损伤和炎症，从血浆中去除ACPSVs后，保护作用消失。通过差速离心分离的EVs表现出ACPSVs的特征。共培养实验表明，ACPSVs可减少缺氧/复氧（H/R）条件下HK-2细胞的凋亡并提高其活力。基因本体（GO）和京都基因与基因组百科全书（KEGG）分析突出了ACPSVs中巨噬细胞迁移抑制因子（MIF）蛋白的关键作用。使用CRISPR/Cas9技术，我们构建了MIF基因敲除的HeLa细胞，以诱导产生缺乏MIF的ACPSVs。当MIF被敲除时，ACPSVs的保护作用显著减弱。转录组测序和染色质免疫沉淀（ChIP）分析表明，MIF通过促进双特异性磷酸酶6（DUSP6）启动子区域的组蛋白H3赖氨酸9三甲基化（H3K9me3）来抑制DUSP6表达，从而激活JNK信号通路。在挽救实验中，用DUSP6抑制剂BCI处理可有效恢复缺乏MIF的ACPSVs的保护功能。
本研究强调了来自rIPC处理大鼠和血清饥饿细胞的ACPSVs通过MIF/DUSP6/JNK信号轴对肾IRI的保护作用，为IRI诱导的急性肾损伤提供了一种潜在的临床治疗策略。
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在线版本包含可在10.1186/s12951-025-03505-9获取的补充材料。
