Alvandi Roshanak, Salimiyan Samira, Moradzad Mohammad, Mohammadi Mobin, Fakhari Shohreh, Rahmani Mohammad Reza
Student Research Committee, Kurdistan University of Medical Sciences, Sanandaj, Iran.
Department of Immunology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran.
Iran J Basic Med Sci. 2024;27(11):1380-1388. doi: 10.22038/ijbms.2024.78183.16905.
Adipose-derived Mesenchymal stem cells (ASCs) have garnered attention for their regenerative potential; therefore, their cellular senescence-related gene expression remains crucial in therapeutic contexts. Nowadays, combination therapies have shown promising results in reducing senescent cells. This study investigated the effects of vitamin C, doxycycline, and azithromycin co-treatment on the key cellular senescence-associated genes in ASCs.
Human ASCs were cultured and treated for 24 hr with vitamin C, doxycycline, azithromycin, and a combination of three drugs. Total RNAs were extracted, and the expression of p21, p16, Nanog, Oct4, and Sox2 genes was assessed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Additionally, cell cycle alterations were analyzed via flow cytometry after treatment with these compounds.
Notably, vitamin C treatment resulted in a significant down-regulation of p21 gene expression (<0.01), implicating the potential role of vitamin C in promoting cell cycle progression. Doxycycline treatment led to a significant up-regulation of p21 and p16 gene expression (<0.05), as it has previously been shown to induce cell cycle arrest. Similarly, azithromycin treatment predominantly increased p21 expression (<0.05). Besides, cell cycle analysis revealed that each compound had changed the distribution of cells across different phases of the cell cycle.
The combined use of all three drugs yielded intricate interactions, suggesting a complex yet promising approach to future research. According to our findings, the major difference in the combination drug-treated group (VDA) can be explained by the neutralizing effect of these three components in the environment.
脂肪来源的间充质干细胞（ASCs）因其再生潜力而备受关注；因此，其细胞衰老相关基因表达在治疗背景下仍然至关重要。如今，联合疗法在减少衰老细胞方面已显示出有前景的结果。本研究调查了维生素C、强力霉素和阿奇霉素联合处理对ASCs中关键细胞衰老相关基因的影响。
培养人ASCs，并用维生素C、强力霉素、阿奇霉素以及三种药物的组合处理24小时。提取总RNA，使用逆转录定量聚合酶链反应（RT-qPCR）评估p21、p16、Nanog、Oct4和Sox2基因的表达。此外，在用这些化合物处理后，通过流式细胞术分析细胞周期变化。
值得注意的是，维生素C处理导致p21基因表达显著下调（<0.01），这表明维生素C在促进细胞周期进程中可能发挥作用。强力霉素处理导致p21和p16基因表达显著上调（<0.05），因为先前已表明它可诱导细胞周期停滞。同样，阿奇霉素处理主要增加p21表达（<0.05）。此外，细胞周期分析表明，每种化合物都改变了细胞在细胞周期不同阶段的分布。
三种药物联合使用产生了复杂的相互作用，这表明未来研究是一种复杂但有前景的方法。根据我们的研究结果，联合药物处理组（VDA）的主要差异可以通过这三种成分在环境中的中和作用来解释。
