Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Key Laboratory of Organ Transplantation, Ministry of Education, Ministry of Public Health, Chinese Academy of Medical Sciences, Wuhan, China.
J Cell Mol Med. 2023 May;27(10):1384-1397. doi: 10.1111/jcmm.17739. Epub 2023 Apr 2.
Human placental mesenchymal stem cells (PMSCs) can prevent liver ischaemia-reperfusion injury (LIRI). However, their therapeutic effects are limited. Therefore, additional research is required to elucidate the mechanisms of PMSC-mediated LIRI prevention and enhance the related therapeutic effects. This study aimed to examine the role of the Lin28 protein in the regulation of glucose metabolism in PMSCs. Further, it explored whether Lin28 could enhance the protective effects of PMSCs against LIRI and investigated the underlying mechanisms. Western blotting was performed to examine Lin28 expression in PMSCs under hypoxic conditions. A Lin28 overexpression construct was introduced into PMSCs, and the effect on glucose metabolism was examined using a glucose metabolism kit. Further, the expression of some proteins involved in glucose metabolism and the PI3K-AKT pathway and the levels of microRNA Let-7a-g were examined using western blots and real-time quantitative PCR, respectively. To examine the relationship between Lin28 and the PI3K-Akt pathway, the effects of AKT inhibitor treatment on the changes induced by Lin28 overexpression were examined. Subsequently, AML12 cells were co-cultured with PMSCs to elucidate the mechanisms via which PMSCs prevent hypoxic injury in liver cells in vitro. Finally, C57BL/6J mice were used to establish a partial warm ischaemia-reperfusion model. The mice received intravenous injections containing PMSCs (control and Lin28-overexpressing PMSCs). Finally, their serum transaminase levels and degree of liver injury were assessed using biochemical and histopathological methods, respectively. Lin28 was upregulated under hypoxic conditions in PMSCs. Lin28 exerted protective effects against hypoxia-induced cell proliferation. Moreover, it increased the glycolytic capacity of PMSCs, allowing PMSCs to produce more energy under hypoxic conditions. Lin28 also activated the PI3K-Akt signalling pathway under hypoxic conditions, and its effects were attenuated by AKT inhibition. Lin28 overexpression could protect cells against LIRI-induced liver damage, inflammation and apoptosis and could also attenuate hypoxia-induced hepatocyte injury. Lin28 enhances glucose metabolism under hypoxic conditions in PMSCs, thereby exerting protective effects against LIRI by activating the PI3K-Akt signalling pathway. Our study is the first to report the potential of genetically modified PMSCs for LIRI treatment.
人胎盘间充质干细胞 (PMSC) 可预防肝缺血再灌注损伤 (LIRI)。然而,它们的治疗效果有限。因此,需要进一步研究阐明 PMSC 介导的 LIRI 预防机制,并增强相关的治疗效果。本研究旨在探讨 Lin28 蛋白在调节 PMSC 葡萄糖代谢中的作用。进一步探讨 Lin28 是否可以增强 PMSC 对 LIRI 的保护作用,并研究其潜在机制。Western blot 检测低氧条件下 PMSC 中 Lin28 的表达。将 Lin28 过表达载体转染入 PMSC,用葡萄糖代谢试剂盒检测葡萄糖代谢的变化。进一步用 Western blot 和实时定量 PCR 检测参与葡萄糖代谢和 PI3K-Akt 通路的某些蛋白的表达以及 microRNA Let-7a-g 的水平。为了研究 Lin28 与 PI3K-Akt 通路的关系,检测 AKT 抑制剂处理对 Lin28 过表达引起的变化的影响。随后,将 AML12 细胞与 PMSC 共培养,以阐明 PMSC 体外预防肝细胞缺氧损伤的机制。最后,使用 C57BL/6J 小鼠建立部分热缺血再灌注模型。小鼠静脉注射含 PMSC(对照和 Lin28 过表达 PMSC)的溶液。最后,通过生化和组织病理学方法分别评估其血清转氨酶水平和肝损伤程度。Lin28 在 PMSC 低氧条件下上调。Lin28 对缺氧诱导的细胞增殖具有保护作用。此外,它增加了 PMSC 的糖酵解能力,使 PMSC 在低氧条件下产生更多的能量。Lin28 还在低氧条件下激活了 PI3K-Akt 信号通路,而 AKT 抑制可减弱其作用。Lin28 过表达可保护细胞免受 LIRI 引起的肝损伤、炎症和细胞凋亡,并减轻低氧诱导的肝细胞损伤。Lin28 增强低氧条件下 PMSC 的葡萄糖代谢,通过激活 PI3K-Akt 信号通路对 LIRI 发挥保护作用。本研究首次报道了基因修饰的 PMSC 治疗 LIRI 的潜力。