Shi Ya-Hui, Li Jun-Qi, Wang Yu-Ying, Wang Ting-Hua, Zuo Zhong-Fu, Liu Xue-Zheng
Liaoning Key Laboratory of Diabetic Cognitive and Perceptive Dysfunction, Jinzhou Medical University, Jinzhou, 121000, China.
Institute of Neuroscience, Kunming Medical University, Kunming, 650500, China.
Curr Stem Cell Res Ther. 2024;19(11):1497-1513. doi: 10.2174/011574888X277276231215110316.
Retinal aging is one of the common public health problems caused by population aging and has become an important cause of acquired vision loss in adults. The aim of this study was to determine the role of human umbilical cord mesenchymal stem cells (hUCMSCs) in delaying retinal ganglion cell (RGC) aging and part of the network of molecular mechanisms involved.
A retinal ganglion cell senescence model was established and treated with UCMSC. Successful establishment of the senescence system was demonstrated using β- galactosidase staining. The ameliorative effect of MSC on senescence was demonstrated using CCK8 cell viability and Annexin V-PI apoptosis staining. The relevant targets of RGC, MSC, and senescence were mainly obtained by searching the GeneCards database. The protein interaction network among the relevant targets was constructed using the String database and Cytoscape, and 10 key target genes were calculated based on the MCC algorithm, based on which Gene ontologies (GO) enrichment and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment were performed. Changes in relevant target genes were detected using real-time fluorescence quantitative PCR and the mechanism of action of UCMSC was determined by RNA interference.
β-galactosidase staining showed that UCMSC significantly reduced the positive results of RGC. The retinal aging process was alleviated. The bioinformatics screen yielded 201 shared genes. 10 key genes were selected by the MCC algorithm, including vascular endothelial growth factor A (VEGFA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), albumin (ALB), interleukin- 6 (IL6), tumor necrosis factor (TNF), tumor protein P53 (TP53), insulin (INS), matrix metalloproteinase 9 (MMP9), epidermal growth factor (EGF), interleukin-1β (IL1B), and enrichment to related transferase activity and kinase activity regulated biological processes involved in oxidative stress and inflammation related pathways. In addition, PCR results showed that all the above molecules were altered in expression after UCMSC involvement.
This experiment demonstrated the role of UCMSC in delaying retinal ganglion cell senescence and further elucidated that UCMSC may be associated with the activation of VEGFA, TP53, ALB, GAPDH, IL6, IL1B, MMP9 genes and the inhibition of INS, EGF, and TNF in delaying retinal senescence.
视网膜老化是人口老龄化引发的常见公共卫生问题之一，已成为成年人后天性视力丧失的重要原因。本研究旨在确定人脐带间充质干细胞（hUCMSCs）在延缓视网膜神经节细胞（RGC）老化中的作用以及部分相关分子机制网络。
建立视网膜神经节细胞衰老模型并用脐带间充质干细胞进行处理。使用β-半乳糖苷酶染色证明衰老系统成功建立。使用CCK8细胞活力检测和膜联蛋白V-碘化丙啶凋亡染色证明间充质干细胞对衰老的改善作用。视网膜神经节细胞、间充质干细胞和衰老的相关靶点主要通过搜索GeneCards数据库获得。使用String数据库和Cytoscape构建相关靶点之间的蛋白质相互作用网络，并基于MCC算法计算出10个关键靶基因，在此基础上进行基因本体（GO）富集和京都基因与基因组百科全书（KEGG）富集。使用实时荧光定量PCR检测相关靶基因的变化，并通过RNA干扰确定脐带间充质干细胞的作用机制。
β-半乳糖苷酶染色显示，脐带间充质干细胞显著降低了视网膜神经节细胞的阳性结果。视网膜老化过程得到缓解。生物信息学筛选产生了201个共享基因。通过MCC算法选择了10个关键基因，包括血管内皮生长因子A（VEGFA）、甘油醛-3-磷酸脱氢酶（GAPDH）、白蛋白（ALB）、白细胞介素-6（IL6）、肿瘤坏死因子（TNF）、肿瘤蛋白P53（TP53）、胰岛素（INS）、基质金属蛋白酶9（MMP9）、表皮生长因子（EGF）、白细胞介素-1β（IL1B），并富集到与氧化应激和炎症相关途径中涉及的相关转移酶活性和激酶活性调节的生物过程。此外，PCR结果显示，脐带间充质干细胞参与后，上述所有分子的表达均发生改变。
本实验证明了脐带间充质干细胞在延缓视网膜神经节细胞衰老中的作用，并进一步阐明脐带间充质干细胞可能与VEGFA、TP53、ALB、GAPDH、IL6、IL1B、MMP9基因的激活以及INS、EGF和TNF的抑制有关，从而延缓视网膜衰老。
