Department of Critical Care Medicine, Strategic Support Force Medical Center, Beijing, China.
Chinese PLA Medical School, Chinese PLA General Hospital, Beijing, China.
FEBS Open Bio. 2022 Mar;12(3):605-615. doi: 10.1002/2211-5463.13365. Epub 2022 Jan 26.
Endothelial dysfunction is a primary pathophysiological change in sepsis. Macrophages are known to interact with vascular endothelial cells during the development of sepsis. Recently, drug delivery based on engineered macrophages was reported as an alternative approach for the management of diseases. Interleukin-10 (IL10) is a well-known anti-inflammatory cytokine, which reduces inflammation and inhibits dysfunction of endothelial cells caused by sepsis. It is currently poorly understood whether genetically modified macrophages with overexpression of IL10 are able to restore endothelial integrity and function at the cellular level. In this study, we used lentiviral vectors to construct RAW264.7 macrophages engineered to overexpress IL10 (IL10-eM) and investigated the effects of the IL10-eM supernatant on LPS-induced endothelial dysfunction using a noncontact coculture system. We found that cotreatment with IL10-eM supernatant significantly attenuates the effects of LPS-induced dysfunction of endothelial cells, including endothelial inflammatory response, endothelial permeability, and apoptosis. In addition, we discovered that LPS-induced downregulation of VE-cadherin and high production of reactive oxygen species were significantly attenuated upon IL10-eM exposure. Furthermore, upregulation of IL6, TNFα, and Bax was decreased after treatment of cells with IL10-eM supernatant. These results demonstrated that supernatant from engineered macrophages genetically modified with IL10 can effectively protect endothelial cells against LPS-induced dysfunction in vitro, suggesting that exosomes from such engineered macrophages may have therapeutic effects against sepsis.
内皮功能障碍是脓毒症的主要病理生理变化。众所周知,巨噬细胞在脓毒症的发展过程中与血管内皮细胞相互作用。最近,基于工程化巨噬细胞的药物输送被报道为治疗疾病的一种替代方法。白细胞介素 10(IL10)是一种众所周知的抗炎细胞因子,可减轻炎症并抑制脓毒症引起的内皮细胞功能障碍。目前尚不清楚过表达 IL10 的基因修饰巨噬细胞是否能够在细胞水平上恢复内皮完整性和功能。在这项研究中,我们使用慢病毒载体构建了过表达 IL10 的 RAW264.7 巨噬细胞(IL10-eM),并使用非接触共培养系统研究了 IL10-eM 上清液对 LPS 诱导的内皮功能障碍的影响。我们发现,IL10-eM 上清液的共处理显著减轻了 LPS 诱导的内皮细胞功能障碍的影响,包括内皮炎症反应、内皮通透性和细胞凋亡。此外,我们发现,IL10-eM 暴露显著减轻了 LPS 诱导的 VE-钙粘蛋白下调和活性氧的大量产生。此外,在用 IL10-eM 上清液处理细胞后,IL6、TNFα 和 Bax 的上调减少。这些结果表明,经基因修饰过表达 IL10 的工程化巨噬细胞的上清液可有效保护内皮细胞免受 LPS 诱导的功能障碍,提示此类工程化巨噬细胞的外泌体可能对脓毒症具有治疗作用。