| 林红杰,王 迪,叶艳艳,熊 宇,税桦桦,黄文静,丁大莲,谭力芯.沉默NHEJ修复通路中关键蛋白Ku70在牙髓干细胞增殖和凋亡中的作用及其机制研究[J].,2021,(5):819-824 |
| 沉默NHEJ修复通路中关键蛋白Ku70在牙髓干细胞增殖和凋亡中的作用及其机制研究 |
| Role and Mechanism of Ku70 in Proliferation and Apoptosis of Dental Pulp Stem Cells |
| 投稿时间：2020-08-07 修订日期：2020-08-31 |
| DOI：10.13241/j.cnki.pmb.2021.05.004 |
| 中文关键词: 非同源重组修复通路 Ku70 牙髓干细胞 |
| 英文关键词: Non homologous recombination repair pathway Ku70 Dental pulp stem cells |
| 基金项目:教育部留学回国人员科研启动基金项目(HG2015-002)；院级军科基金项目(SWH2016JSTSYB-40N) |
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| 中文摘要: |
| 摘要 目的：研究沉默非同源重组修复（non-homologous endjoining，NHEJ）通路中关键蛋白Ku70在牙髓干细胞增殖和凋亡中的作用，分析其机制。方法：提取健康恒牙牙髓组织，进行牙髓干细胞培养。采用脂多糖诱导人牙髓干细胞，分为对照组、阴性对照组、脂多糖组、沉默组和沉默+脂多糖组。观察Ku70免疫组化情况，进行细胞增殖、细胞凋亡实验，检测γ-H2A.X、Ku70、X线修复交叉互补基因4（X-ray repair cross complementary gene 4，Xrcc4）、Rad51 mRNA、Cleaved Caspase-3、p-p38水平。结果：与沉默组相比，对照组、阴性对照组、脂多糖组、沉默+脂多糖组各时间段牙髓干细胞增殖降低；沉默+脂多糖牙髓干细胞增殖高于脂多糖组（P＜0.05）。随时间延长，脂多糖组牙髓干细胞增殖不断降低，其他四组牙髓干细胞增殖不断升高，其中在第5 d变化最明显。第5d，与沉默组相比，对照组、阴性对照组牙髓干细胞凋亡率、γ-H2A.X、Rad51 mRNA，Cleaved Caspase-3降低，p-p38升高；脂多糖组、沉默+脂多糖组各项指标较高，p-p38降低；对照组、阴性对照组、脂多糖组、沉默+脂多糖组Ku70、Xrcc4 mRNA降低（P＜0.05）。沉默+脂多糖组牙髓干细胞凋亡率、γ-H2A.X、Rad51 mRNA，Cleaved Caspase-3低于脂多糖组，p-p38高于脂多糖组（P＜0.05）。结论：沉默Ku70能促进脂多糖诱导的牙髓干细胞增殖，抑制其凋亡，其可能与γ-H2A.X、Rad51 mRNA表达降低，Ku70，Xrcc4升高有关。 |
| 英文摘要: |
| ABSTRACT Objective: To study the role of the key protein Ku70 in the silent non-homologous endjoining (NHEJ) pathway in the proliferation and apoptosis of dental pulp stem cells and to analyze its mechanism. Methods: The pulp tissue of healthy permanent teeth was extracted and cultured with pulp stem cells.LPS was used to induce human dental pulp stem cells, which were divided into control group, negative control group, LPS group, silencing group and silencing + LPS group. The immunohistochemistry of Ku70 was observed,cell proliferation and apoptosis experiments were conducted to detect the levels of gourie-h21. X, Ku70, X-ray repair cross complementary gene 4 (Xrcc4), Rad51 mRNA, Cleaved caspase-3 and p-p38. Results: Compared with the silence group, the proliferation of dental pulp stem cells decreased in the control group, the negative control group, the LPS group and the silence + LPS group. The proliferation of dental pulp stem cells in the silencing + LPS group was higher than that in the LPS group (P<0.05). With the extension of time, the proliferation of dental pulp stem cells in the LPS group decreased continuously, while the proliferation of dental pulp stem cells in the other four groups increased continuously, among which the change was most obvious on the 5th day. On the 5th day, compared with the silence group, the apoptosis rate of dental pulp stem cells, lap-h21. X, Rad51 mRNA, and Cleaved caspase-3 in the control group and the negative control group decreased, but p-p38 increased.In the LPS group and the silencing + LPS group, all indexes were higher and p-p38 was lower. The levels of Ku70 and Xrcc4 mRNA were decreased in the control group, the negative control group, the LPS group, and the silence + LPS group (P<0.05). The apoptosis rate of dental pulp stem cells, lap-h2a2.X, Rad51 mRNA, Cleaved caspase-3 in the silencing + LPS group were lower than those in the LPS group, while p-p38 was higher than those in the LPS group (P<0.05). Conclusion: Silencing Ku70 could promote the proliferation of lps-induced dental pulp stem cells and inhibit their apoptosis, which might be related to the decreased expression of γ-H2A.X and Rad51 mRNA, and the increase of Ku70 and Xrcc4. |
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