Mesenchymal stem/stromal cells (MSCs) have been identified in almost all adult human tissues and been used in numerous clinical trials for a variety of diseases. Studies have shown that MSCs would undergo cellular senescence when cultured over a long term, which is brought on by increased epigenetic modifications, including DNA methylation. However, the mechanism of MSCs senescence is not well studied. In this study, the effects of RG108, a DNA methyltransferase inhibitor (DNMTi), on senescence, apoptosis, and pluripotency gene expressions in porcine bone marrow (pBM)-MSCs were investigated. First, we determined the optimized dose and time of RG108 treatment in pBM-MSCs to be 10 μM for 48 hours, respectively. Under these conditions, the pluripotency genes (NANOG, POU5F1), the anti-senescence genes (TERT, bFGF), and the anti-apoptosis gene (BCL2) were increased, whereas the apoptotic gene (BAX) was decreased. RG108 protected against apoptosis when pBM-MSC induces apoptosis with H₂O₂ for 1.5 hours. We also found that RG108 significantly induced the expression of NANOG and POU5F1 by decreasing DNA methylation in gene promoter regions. These results indicate that an optimized dose of RG108 may promote the pluripotency-related character of pBM-MSCs through improving cellular anti-senescence, anti-apoptosis, and pluripotency, which provide a better cell origin for stem cell therapy.

中文翻译：

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RG108对DNA甲基转移酶的抑制促进了猪骨髓间充质干细胞的多能性相关特征。

间充质干/基质细胞（MSCs）已在几乎所有成人组织中得到鉴定，并已用于多种疾病的众多临床试验中。研究表明，长期培养后，MSC会发生细胞衰老，这是由表观遗传修饰（包括DNA甲基化）增加所引起的。然而，对MSCs衰老的机制还没有很好的研究。在这项研究中，研究了RG108，一种DNA甲基转移酶抑制剂（DNMTi）对猪骨髓（pBM）-MSCs衰老，凋亡和多能性基因表达的影响。首先，我们确定pBM-MSC中RG108治疗的最佳剂量和时间分别为10μM，持续48小时。在这些条件下，多能性基因（NANOG，POU5F1），抗衰老基因（TERT，bFGF）和抗凋亡基因（BCL2）增加，而凋亡基因（BAX）减少。当pBM-MSC用H ₂ O ₂诱导1.5小时凋亡时，RG108可以防止凋亡。我们还发现，RG108可通过减少基因启动子区域的DNA甲基化来显着诱导NANOG和POU5F1的表达。这些结果表明优化剂量的RG108可以通过改善细胞抗衰老，抗凋亡和多能性来促进pBM-MSC的多能性相关特性，这为干细胞治疗提供了更好的细胞来源。