骨髓间充质干细胞来源的包含 miR-539-5p 的外泌体通过 NLRP3/caspase-1 信号通路抑制细胞焦亡，从而减轻炎症性肠病。
Bone marrow mesenchymal stem cells-derived exosomes containing miR-539-5p inhibit pyroptosis through NLRP3/caspase-1 signalling to alleviate inflammatory bowel disease.
机构信息
Department of Anorectum, The Affiliated Hospital of Qingdao University, No. 16 Jiangsu Road, Qingdao, 266000, Shandong Province, People's Republic of China.
Department of Gynecology, Qingdao Hospital of Traditional Chinese Medicine, Qingdao, 266100, Shandong Province, People's Republic of China.
出版信息
Inflamm Res. 2022 Aug;71(7-8):833-846. doi: 10.1007/s00011-022-01577-z. Epub 2022 May 30.
BACKGROUND
Exosomes derived from bone mesenchymal stem cells (BMSCs) are potential candidates for inflammatory bowel disease (IBD) treatment. The present study investigated the therapeutic effect and potential mechanism of BMSCs-derived exosomes on pyroptosis in IBD.
METHODS
We induced IBD in mice and cell models through dextran sulfate sodium (DSS) and LPS, respectively. The mRNA and protein expression levels were assessed by qRT-PCR, Western blotting, IF and IHC. The concentrations of IL-1β, IL-18 and TNFα were assessed using ELISA. ROS levels were determined using DCFH-DA staining. Cell proliferation of mIECs was analysed using an MTT assay. In addition, a flow cytometry assay was performed to detect pyroptosis. Finally, the binding relationship between miR-539-5p and NLRP3 was verified by a dual luciferase reporter gene assay.
RESULTS
Our results revealed that intraperitoneal injection of BMSCs-derived exosomes inhibited DSS-induced pyroptosis as well as IBD symptoms in mice. In addition, BMSCs-derived exosome treatment suppressed pyroptosis, ROS levels and the concentrations of proinflammatory cytokines (IL-1β, IL-18 and TNFα) in LPS-treated mIECs in a miR-539-5p-dependent manner. Further research found that miR-539-5p suppressed NLRP3 expression in mIECs by directly targeting NLRP3. As expected, pyroptosis in LPS-treated mIECs was significantly reduced by NLRP3 knockdown. In addition, NLRP3 silencing restored the inhibitory effect of exosomes derived from BMSCs transfected with miR-539-5p inhibitor on pyroptosis in LPS-treated mIECs.
CONCLUSION
The present study demonstrated that BMSCs-derived exosomal miR-539-5p suppresses pyroptosis through NLRP3/caspase-1 signalling to inhibit IBD progression.
背景
骨间充质干细胞（BMSC）衍生的外泌体是治疗炎症性肠病（IBD）的潜在候选物。本研究探讨了 BMSC 衍生的外泌体对 IBD 细胞焦亡的治疗作用及潜在机制。
方法
通过葡聚糖硫酸钠（DSS）和 LPS 分别在小鼠和细胞模型中诱导 IBD。通过 qRT-PCR、Western blot、IF 和 IHC 评估 mRNA 和蛋白表达水平。ELISA 法检测 IL-1β、IL-18 和 TNFα 的浓度。DCFH-DA 染色法测定 ROS 水平。MTT 法分析 mIECs 的细胞增殖。此外，通过流式细胞术检测细胞焦亡。最后，通过双荧光素酶报告基因实验验证 miR-539-5p 与 NLRP3 之间的结合关系。
结果
结果表明，BMSC 衍生的外泌体腹腔注射抑制了 DSS 诱导的小鼠细胞焦亡和 IBD 症状。此外，BMSC 衍生的外泌体处理以 miR-539-5p 依赖的方式抑制 LPS 处理的 mIECs 中的细胞焦亡、ROS 水平和促炎细胞因子（IL-1β、IL-18 和 TNFα）的浓度。进一步的研究发现，miR-539-5p 通过直接靶向 NLRP3 抑制 mIECs 中 NLRP3 的表达。正如预期的那样，LPS 处理的 mIECs 中的细胞焦亡明显减少。此外，NLRP3 沉默恢复了 miR-539-5p 抑制剂转染的 BMSC 衍生的外泌体对 LPS 处理的 mIECs 中细胞焦亡的抑制作用。
结论
本研究表明，BMSC 衍生的外泌体 miR-539-5p 通过 NLRP3/caspase-1 信号通路抑制细胞焦亡，从而抑制 IBD 的进展。
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