Jiang Meng, Wan Shengxia, Dai Xiaoyong, Ye Youwen, Hua Wei, Ma Guoguang, Pang Xiufeng, Wang Huanhuan, Shi Bin
Department of Emergency Intensive Care Unit, Yangpu Hospital, Tongji University, Shanghai 200090, China.
Department of Neurology, The Fourth Affiliated Hospital of Jiangsu University, Zhenjiang 212000, China.
Open Med (Wars). 2022 Jul 20;17(1):1308-1317. doi: 10.1515/med-2022-0520. eCollection 2022.
This study aimed to investigate whether ghrelin affected the autophagy and inflammatory response of intestinal intraepithelial lymphocytes (IELs) by regulating the NOD2/Beclin-1 pathway in an intestinal ischemia-reperfusion (I/R) injury model. Twenty hours after implementing the intestinal I/R injury rat model, the small intestine and both lungs were collected for histological analysis. The morphological changes in the intestinal mucosa epithelium and lung tissues were evaluated using hematoxylin-eosin staining. The activity of autophagic vacuoles and organ injury were evaluated using electron microscopy. The cytokine levels (IL-10 and TNF-α) in IEL cells and lung tissue were determined using enzyme-linked immunosorbent assay. RT-qPCR and western blot assays were conducted to check the NOD2, Beclin-1, and ATG16 levels. Ghrelin relieved the I/R-induced destruction of the intestinal mucosa epithelium and lung tissues. Moreover, ghrelin enhanced autophagy in the intestinal epithelium and lungs of I/R rats. In addition, the levels of autophagy-associated proteins (Beclin-1, ATG16, and NOD2) were higher in the ghrelin treatment group than in rats with I/R. Ghrelin reduced significantly the IL-10 and TNF-α levels. However, these changes were reversed by the NOD2 antagonist. In conclusion, ghrelin may relieve I/R-induced acute intestinal mucosal damage, autophagy disorder, and inflammatory response in IELs by regulating the NOD2/Beclin-1 pathway.
本研究旨在探讨在肠缺血再灌注（I/R）损伤模型中，胃饥饿素是否通过调节NOD2/Beclin-1信号通路影响肠上皮内淋巴细胞（IELs）的自噬和炎症反应。建立大鼠肠I/R损伤模型20小时后，收集小肠和双肺进行组织学分析。采用苏木精-伊红染色评估肠黏膜上皮和肺组织的形态学变化。采用电子显微镜评估自噬泡活性和器官损伤情况。采用酶联免疫吸附测定法测定IEL细胞和肺组织中的细胞因子水平（IL-10和TNF-α）。进行RT-qPCR和蛋白质免疫印迹分析以检测NOD2、Beclin-1和ATG16的水平。胃饥饿素减轻了I/R诱导的肠黏膜上皮和肺组织破坏。此外，胃饥饿素增强了I/R大鼠肠上皮和肺中的自噬。另外，胃饥饿素治疗组中自噬相关蛋白（Beclin-1、ATG16和NOD2）的水平高于I/R大鼠。胃饥饿素显著降低了IL-10和TNF-α水平。然而，这些变化被NOD2拮抗剂逆转。总之，胃饥饿素可能通过调节NOD2/Beclin-1信号通路减轻I/R诱导的急性肠黏膜损伤、自噬紊乱和IELs中的炎症反应。
