Ye Kui, Peng Qiu-Feng, Zhang Zeng-Guang, Li Xiang, Xue Hong-Bo, Guo Hua, Hei Hao-Yang, Wang Hao
Department of Vascular Surgery, Tianjin Fourth Central Hospital, Tianjin Medical University, Tianjin, China; Department of Vascular Surgery, Tianjin Fourth Central Hospital, Tianjin University, Tianjin, China.
Department of General Surgery, Tianjin Medical University General Hospital, Tianjin, China.
Int Immunopharmacol. 2025 Sep 3;165:115481. doi: 10.1016/j.intimp.2025.115481.
Intestinal ischemia/reperfusion injury (IRI) is a severe clinical condition associated with high morbidity and mortality. Despite advances in understanding the pathophysiology of IRI, effective therapeutic strategies remain limited. Adipose stem cell (ADSC)-derived exosomes (Exo) have been proven to be appropriate candidates for IRI through the anti-inflammatory effects. In this study, we investigated whether resveratrol (RSV)-primed ADSC-derived exosomes (RSV-primed Exo) could exert superior effects on alleviating intestinal IRI, and attempted to elucidate the underlying mechanism.
For the evaluation of the therapeutic effect of the RSV-primed Exo on intestinal IRI, in vitro and in vivo studies were performed. IRI model was established in male C57BL/6 mice by clamping the superior mesenteric artery for 45 min followed by reperfusion. Histopathological analysis, intestinal barrier, the proportion of T cells, and macrophage polarization were investigated. RSV-primed Exo mediated immunosuppressive activity was investigated by bone marrow-derived macrophages (BMDM) co-culture assay in vitro. Flow cytometry determined macrophage polarization. ELISA assays detected the function of macrophages. Furthermore, the role of the NF-κB pathway in RSV-primed Exo induced macrophage polarization was also elucidated.
In vivo modeling in intestinal IRI showed that RSV pretreatment enhanced the therapeutic efficacy of ADSC-derived exosomes. Compared with Untreated and Exo-treated groups, RSV-primed Exo effectively attenuated pathological damage of intestine characterized by severe edema of the mucosal villi and the infiltration of inflammatory cells. Exo therapy conspicuously restored the downregulated expression of ZO-1, Occludin and Claudin-1 proteins in intestinal IRI mice and the administration of RSV-primed Exo further enhanced this restorative effect. Surprisingly, RSV-primed Exo significantly decreased the populations of M1 macrophages but increased the proportions of M2 macrophages in the intestine. Furthermore, RSV-primed Exo markedly reduced the levels of proinflammatory cytokines (IL-1β, IL-6, and TNF-α) and increased anti-inflammatory factor (IL-10) level in intestine. RSV-primed Exo improved macrophage immunoregulatory function associated with the NF-κB pathway, which exerted a potent therapeutic effect against intestinal IRI.
We demonstrated that RSV-primed Exo effectively suppress intestinal barrier dysfunctions and the dysregulation of mucosal immune responses. The results highlight that RSV-primed Exo could be novel and promising candidate for prevention of intestinal IRI.
肠缺血/再灌注损伤（IRI）是一种严重的临床病症，发病率和死亡率都很高。尽管在理解IRI的病理生理学方面取得了进展，但有效的治疗策略仍然有限。脂肪干细胞（ADSC）来源的外泌体（Exo）已被证明通过抗炎作用是IRI的合适候选物。在本研究中，我们调查了白藜芦醇（RSV）预处理的ADSC来源的外泌体（RSV预处理的Exo）是否能在减轻肠道IRI方面发挥更优效果，并试图阐明其潜在机制。
为评估RSV预处理的Exo对肠道IRI的治疗效果，进行了体外和体内研究。通过夹闭雄性C57BL/6小鼠的肠系膜上动脉45分钟，然后再灌注，建立IRI模型。进行组织病理学分析、肠道屏障、T细胞比例和巨噬细胞极化研究。通过体外骨髓来源的巨噬细胞（BMDM）共培养试验研究RSV预处理的Exo介导的免疫抑制活性。流式细胞术测定巨噬细胞极化。ELISA检测巨噬细胞功能。此外，还阐明了NF-κB通路在RSV预处理的Exo诱导巨噬细胞极化中的作用。
肠道IRI的体内建模显示，RSV预处理增强了ADSC来源外泌体的治疗效果。与未处理组和Exo处理组相比，RSV预处理的Exo有效减轻了以粘膜绒毛严重水肿和炎症细胞浸润为特征的肠道病理损伤。Exo疗法显著恢复了肠道IRI小鼠中紧密连接蛋白1（ZO-1）、闭合蛋白（Occludin）和Claudin-1蛋白下调的表达，而给予RSV预处理的Exo进一步增强了这种恢复作用。令人惊讶的是，RSV预处理的Exo显著减少了肠道中M1巨噬细胞的数量，但增加了M2巨噬细胞的比例。此外，RSV预处理的Exo显著降低了肠道中促炎细胞因子（白细胞介素-1β、白细胞介素-6和肿瘤坏死因子-α）的水平，并增加了抗炎因子（白细胞介素-10）的水平。RSV预处理的Exo改善了与NF-κB通路相关的巨噬细胞免疫调节功能，对肠道IRI发挥了强大的治疗作用。
我们证明了RSV预处理的Exo有效抑制肠道屏障功能障碍和粘膜免疫反应失调。结果表明，RSV预处理的Exo可能是预防肠道IRI的新型且有前景的候选物。
