Duangprom Supawadee, Kheolamai Pakpoom, Tantrawatpan Chairat, Manochantr Sirikul
Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Pathumthani, 12120, Thailand.
Center of Excellence in Stem Cell Research and Innovation, Thammasat University, Pathumthani, 12120, Thailand.
Sci Rep. 2025 Jul 2;15(1):22512. doi: 10.1038/s41598-025-06454-3.
Chronic hyperglycemia is recognized as an important contributor to chronic inflammation, oxidative stress, and organ dysfunction that causes serious complications in diabetes and aging. This study investigates the effects of elevated glucose levels on human placenta-derived mesenchymal stem cells (hP-MSCs), especially since these cells hold a promise for tissue engineering and regenerative medicine due to their multipotency and self-renewal capabilities. hP-MSCs were treated with 10-40 mM D-glucose to study the effects of high glucose on hP-MSCs functions. hP-MSCs viability and proliferation were determined by using thiazolyl blue tetrazolium bromide (MTT), cell cycle analysis, and senescence assays. The migration and osteogenic differentiation capacity were also determined by migration assay, alkaline phosphatase (ALP) activity assay, and Alizarin Red S staining. Quantitative real-time RT-PCR, Western blot, and Nanostring® nCounter assay were performed to study the effect of high glucose on the expression levels of genes involved in various aspects of hP-MSCs functions. The results demonstrated that high glucose significantly inhibited proliferation and cell cycle progression of hP-MSCs at the G1/S phase and induced replicative senescence in hP-MSCs possibly by decreasing the expression of proliferation-promoting genes, CCND1 and LMNB1, and increasing the expression of several senescence-associated proteins, p16, p21, and p53. Furthermore, high glucose also inhibited the migration and osteogenic differentiation of hP-MSCs, possibly by suppressing the expression of SDF1, CXCR4, RUNX2, OSX, OCN, and COL1A. The additional Nanostring analysis also showed that high glucose significantly affects multiple genes involved in inflammation, DNA repair, autophagy, and oxidative stress response in hP-MSCs. This study provides significant insights into the wide-ranging effects of high glucose on the expression of the hP-MSCs genes that affect various aspects of its function, including proliferation, viability, senescence, oxidative stress response, and DNA repair, highlighting its implications for regenerative medicine in the context of diabetes and metabolic disorders.
慢性高血糖被认为是导致慢性炎症、氧化应激和器官功能障碍的重要因素，这些会在糖尿病和衰老过程中引发严重并发症。本研究调查了葡萄糖水平升高对人胎盘来源间充质干细胞（hP-MSCs）的影响，特别是鉴于这些细胞因其多能性和自我更新能力，在组织工程和再生医学方面具有应用前景。用10 - 40 mM D-葡萄糖处理hP-MSCs，以研究高糖对hP-MSCs功能的影响。通过噻唑蓝四氮唑溴盐（MTT）、细胞周期分析和衰老检测来测定hP-MSCs的活力和增殖。迁移和骨生成分化能力也通过迁移检测、碱性磷酸酶（ALP）活性检测和茜素红S染色来确定。进行定量实时RT-PCR、蛋白质印迹和Nanostring® nCounter检测，以研究高糖对参与hP-MSCs功能各个方面的基因表达水平的影响。结果表明，高糖显著抑制hP-MSCs在G1/S期的增殖和细胞周期进程，并可能通过降低增殖促进基因CCND1和LMNB1的表达，以及增加几种衰老相关蛋白p16、p21和p53的表达，诱导hP-MSCs发生复制性衰老。此外，高糖还可能通过抑制SDF1、CXCR4、RUNX2、OSX、OCN和COL1A的表达，抑制hP-MSCs的迁移和骨生成分化。额外的Nanostring分析还表明，高糖显著影响hP-MSCs中涉及炎症、DNA修复、自噬和氧化应激反应的多个基因。本研究为高糖对hP-MSCs基因表达的广泛影响提供了重要见解，这些基因影响其功能的各个方面，包括增殖、活力、衰老、氧化应激反应和DNA修复，突出了其在糖尿病和代谢紊乱背景下对再生医学的意义。
