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首页> 外文期刊>Cell and Tissue Research >Repeated lipopolysaccharide stimulation promotes cellular senescence in human dental pulp stem cells (DPSCs).
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Repeated lipopolysaccharide stimulation promotes cellular senescence in human dental pulp stem cells (DPSCs).

机译：重复的脂多糖刺激促进人牙髓干细胞（DPSC）中的细胞衰老。

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Dental pulp stem cells (DPSCs) are a type of mesenchymal stem cell (MSC) characterized by multi-lineage differentiation making it an attractive choice for tissue regeneration. However, before DPSCs can be used for cell-based therapy, we have to understand their biological properties in response to intrinsic and extrinsic stimuli such as lipopolysaccharide (LPS). DPSCs were therefore stimulated with LPS and senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining, with cell number and cell-cycle arrest being examined by BrdU assay and flow cytometry, respectively. The morphology of DPSCs was characterized by their flat shape, increased size and increased SA-β-gal activity after repeated stimulation (3 or 6 times) with LPS. Reactive oxygen species (ROS) staining showed that the number of ROS-stained cells and the DCFH fluorescent level were higher in the LPS-treated DPSCs compared with those in the untreated DPSCs. Protein and mRNA expression levels of γ-H2A.X and p16(INK4A) were also increased in DPSCs with repeated LPS stimulation. We found that the LPS bound with Toll-like receptor 4 (TLR4) and that TLR4 signaling accounted for p16(INK4A) expression. Further results indicated that the senescence of DPSCs stimulated repeatedly with LPS was reversed by p16(INK4A) short interfering RNA. The DNA damage response and p16(INK4A) pathways might be the main mediators of DPSC senescence induced by repeated LPS stimulation. Thus, DPSCs tend to undergo senescence after repeated activation, implying that DPSC senescence starts after many inflammatory challenges. Ultimately, these findings should lead to a better understanding of DPSC-based clinical therapy.
机译：牙髓干细胞（DPSC）是一种间充质干细胞（MSC），其特征是多谱系分化，使其成为组织再生的诱人选择。但是，在DPSC可以用于基于细胞的治疗之前，我们必须了解其对内在和外在刺激（例如脂多糖（LPS））的生物学特性。因此，用LPS刺激DPSCs，并通过衰老相关的β-半乳糖苷酶（SA-β-gal）染色来评估衰老，并分别通过BrdU测定法和流式细胞术检查细胞数目和细胞周期停滞。用LPS反复刺激（3或6次）后，DPSC的形态特征为扁平形状，尺寸增加和SA-β-gal活性增加。活性氧（ROS）染色显示，与未处理的DPSC相比，LPS处理的DPSC中ROS染色的细胞数和DCFH荧光水平更高。反复LPS刺激的DPSCs中γ-H2A.X和p16（INK4A）的蛋白质和mRNA表达水平也增加。我们发现LPS与Toll样受体4（TLR4）绑定，并且TLR4信号传导占p16（INK4A）的表达。进一步的结果表明，LPS反复刺激的DPSC的衰老被p16（INK4A）短干扰RNA逆转。 DNA损伤反应和p16（INK4A）途径可能是反复LPS刺激诱导DPSC衰老的主要介质。因此，DPSCs在反复激活后趋于衰老，这意味着DPSC衰老在许多炎症挑战后开始。最终，这些发现将使人们对基于DPSC的临床疗法有更好的了解。

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