Gao Yao, He Min, Bian Cong-Wen, Yu Rui, Luo Jia-Jiao, Xiang Yin-Ming, Yang Yun-Xin, Huang Han-Fei, Zeng Zhong
The Organ Transplantation Center, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China.
Department of Hepatic, Guangzhou Eighth People's Hospital, Guangzhou Medical University, Guangzhou 510440, Guangdong Province, China.
World J Gastroenterol. 2025 Sep 7;31(33):108653. doi: 10.3748/wjg.v31.i33.108653.
Hepatic ischaemia-reperfusion injury (HIRI) is an unavoidable process in liver transplantation, where apoptosis plays a critical role. Human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-exos), which constitute a cell-free therapeutic approach, have garnered extensive attention in alleviating HIRI. However, the potential of hucMSC-exos in mitigating apoptosis and their underlying mechanisms remain largely unknown.
To investigate the effects of hucMSC-exos on apoptosis after HIRI and explore the underlying mechanisms.
The therapeutic effects of hucMSC-exos on HIRI and hypoxia/reoxygenation injury in L02 cells were investigated. RNA sequencing was used to detect differentially expressed genes in L02 cells after hucMSC-exo treatment, and the expression of apoptosis markers in L02 cells was analyzed. MicroRNA (miRNA) sequencing was performed to analyse the miRNA expression profiles of hucMSC-exos and L02 cells after hucMSC-exo treatment. Through a miRNA-mRNA integrated analysis, candidate miRNAs and their regulated target genes were identified. We subsequently studied the roles of these candidate miRNAs in mouse HIRI and L02 cell hypoxia/reoxygenation injury.
Fluorescence confocal microscopy revealed that hucMSC-exos effectively homed to the liver and were taken up by hepatocytes, likely due to the presence of anti-very late antigen-4 and anti-lymphocyte function-associated antigen-1 on the surface of hucMSC-exos. HucMSC-exos alleviate hepatocyte damage by inhibiting apoptosis. Specifically, let-7i-5p within hucMSC-exos inhibited the expression of the factor-related apoptosis ligand protein in L02 cells, leading to the upregulation of B-cell lymphoma-2 and the downregulation of B-cell lymphoma-2-associated X protein and cysteinyl aspartate specific proteinase-3, thereby inhibiting L02 cell apoptosis and enhancing cell proliferation activity. The overexpression of let-7i-5p effectively enhanced the antiapoptotic effects of hucMSC-exos both and .
Our findings indicate that hucMSC-exos alleviate HIRI by inhibiting apoptosis. We demonstrated that hucMSC-exos target apoptosis in L02 cells and mediate the let-7i-5p/factor-related apoptosis ligand pathway, thereby ameliorating HIRI. This study provides new insights into the role of hucMSC-exos in hepatocyte apoptosis and highlights the potential of hucMSC-exos as a therapeutic strategy for HIRI.
肝缺血再灌注损伤（HIRI）是肝移植中不可避免的过程，其中细胞凋亡起关键作用。人脐带间充质干细胞衍生的外泌体（hucMSC-exos）作为一种无细胞治疗方法，在减轻HIRI方面受到广泛关注。然而，hucMSC-exos在减轻细胞凋亡方面的潜力及其潜在机制仍 largely未知。
研究hucMSC-exos对HIRI后细胞凋亡的影响并探讨其潜在机制。
研究了hucMSC-exos对HIRI以及L02细胞缺氧/复氧损伤的治疗作用。采用RNA测序检测hucMSC-exo处理后L02细胞中差异表达基因，并分析L02细胞中凋亡标志物的表达。进行微小RNA（miRNA）测序以分析hucMSC-exos及hucMSC-exo处理后L02细胞的miRNA表达谱。通过miRNA-mRNA综合分析，鉴定候选miRNA及其调控的靶基因。随后我们研究了这些候选miRNA在小鼠HIRI和L02细胞缺氧/复氧损伤中的作用。
荧光共聚焦显微镜显示hucMSC-exos有效归巢至肝脏并被肝细胞摄取，这可能是由于hucMSC-exos表面存在抗极晚期抗原-4和抗淋巴细胞功能相关抗原-1。HucMSC-exos通过抑制细胞凋亡减轻肝细胞损伤。具体而言，hucMSC-exos中的let-7i-5p抑制L02细胞中因子相关凋亡配体蛋白的表达，导致B细胞淋巴瘤-2上调，B细胞淋巴瘤-2相关X蛋白和半胱氨酸天冬氨酸特异性蛋白酶-3下调，从而抑制L02细胞凋亡并增强细胞增殖活性。let-7i-5p的过表达在体内和体外均有效增强了hucMSC-exos的抗凋亡作用。
我们的研究结果表明hucMSC-exos通过抑制细胞凋亡减轻HIRI。我们证明hucMSC-exos靶向L02细胞中的细胞凋亡并介导let-7i-5p/因子相关凋亡配体途径，从而改善HIRI。本研究为hucMSC-exos在肝细胞凋亡中的作用提供了新见解，并突出了hucMSC-exos作为HIRI治疗策略的潜力。
