Institute of Neuroscience, Kunming Medical University, Kunming, 650500, China.
Animal Center, Kunming Medical University, Kunming, 650500, China.
Biochem Genet. 2024 Oct;62(5):4046-4066. doi: 10.1007/s10528-023-10487-6. Epub 2024 Jan 25.
To investigate the effect and potential mechanism of human-derived urine stem cells (hUSCs) in inhibiting retinal aging by using experimental and bioinformatics. Retinal pigment epithelial cells cultured in vitro, which were randomly divided into normal group, aging group and supernatant of hUSCs group. Cell counting kit-8 detection, senescence-related β-galactosidase, and Annexin V/PI staining were performed to detect cell viability, senescence, and apoptosis. Subsequently, bioinformatics methods were used to explore the underlying mechanisms, in which, targets both hUSCs and aging retina-related targets were obtained from GeneCards. Then, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes enrichment analysis, and protein-protein interaction network were analysis, and the expressional level of hub gene was validated by q-PCR. Supernatant addition of hUSCs promoted markedly cellular proliferation, improved viability and inhibited senescence and apoptosis in vitro. A total of 1476 hUSCs-related targets (Relevance score > 20), 692 retinal disease-related targets, and 732 targets related to disease of aging were selected from GeneCards database, and 289 common targets of hUSCs against aging retina were confirmed through Venn analysis. Enrichment analysis demonstrated that hUSCs might exert its anti-apoptosis efficacy in multiple biological processes, including oxidative stress, inflammation and apoptosis, and core targets were associated with HIF-1, MAPK and PI3K-Akt signal. hUSCs inhibited retinal senescence by regulating multiply targets and signaling pathways, of these, HIF-1, MAPK, and PI3K may be important candidates.
为了通过实验和生物信息学来研究人源性尿干细胞（hUSCs）抑制视网膜衰老的作用和潜在机制，我们将体外培养的视网膜色素上皮细胞随机分为正常组、衰老组和 hUSCs 上清组。通过细胞计数试剂盒-8 检测、衰老相关β-半乳糖苷酶、Annexin V/PI 染色来检测细胞活力、衰老和凋亡。随后，我们使用生物信息学方法来探索潜在机制，其中，从 GeneCards 中获得了 hUSCs 和衰老视网膜相关的靶点。然后，进行基因本体论、京都基因与基因组百科全书富集分析和蛋白质-蛋白质相互作用网络分析，并通过 q-PCR 验证了核心基因的表达水平。hUSCs 上清液的添加显著促进了细胞增殖，提高了细胞活力，抑制了体外衰老和凋亡。从 GeneCards 数据库中选择了 1476 个 hUSCs 相关靶点（相关性评分>20）、692 个视网膜疾病相关靶点和 732 个与衰老相关的靶点，通过 Venn 分析确认了 289 个 hUSCs 对抗衰老视网膜的共同靶点。富集分析表明，hUSCs 可能通过多种生物学过程发挥其抗凋亡作用，包括氧化应激、炎症和凋亡，核心靶点与 HIF-1、MAPK 和 PI3K-Akt 信号有关。hUSCs 通过调节多种靶点和信号通路来抑制视网膜衰老，其中 HIF-1、MAPK 和 PI3K 可能是重要的候选靶点。
