过表达ADAM10通过调控Notch信号通路对牙周膜干细胞成骨分化的影响

doi:10.13701/j.cnki.kqyxyj.2021.05.018

摘要/Abstract

摘要： 目的:探讨解整合素金属蛋白酶10(ADAM10)调控Notch信号通路对人牙周膜干细胞(hPDLSCs)成骨分化的影响。方法:体外培养hPDLSCs并对其进行成骨分化诱导,RT-PCR检测成骨诱导第0、7、14天细胞中ADAM10 mRNA表达水平。采用ADAM10过表达慢病毒感染hPDLSCs,并将细胞分为空白对照组、空载组和ADAM10过表达组,分别采用RT-PCR和Western blot检测感染后hPDLSCs中ADAM10 mRNA和蛋白表达水平,CCK-8检测各组细胞增值情况。感染的hPDLSCs经成骨诱导分化后,茜素红染色观察各组细胞矿化结节生成情况,RT-PCR检测各组细胞中成骨相关基因ALP、Runx2、OCN以及Notch信号通路相关基因Notch1、NICD、Hes1的mRNA表达水平,Western blot检测Notch信号通路相关蛋白Notch1、NICD和Hes1的蛋白表达水平。结果:hPDLSCs成骨诱导分化的第0、7、14天过程中,ADAM10 mRNA表达水平逐渐降低(P<0.05)。感染ADAM10过表达慢病毒后,hPDLSCs中ADAM10 mRNA和蛋白表达水平显著增加(P<0.05),同时细胞增殖能力明显增强(P<0.05)。成骨诱导分化后,与对照组和空载组比较,ADAM10过表达组矿化结节形成量、成骨相关基因ALP、Runx2、OCN的mRNA表达水平以及Notch信号通路相关基因Notch1、NICD、Hes1的mRNA和蛋白表达水平均显著降低(P<0.05)。结论:过表达ADAM10可促进hPDLSCs增殖并抑制其成骨分化,其机制可能与Notch信号通路的激活有关。

关键词: 解整合素金属蛋白酶10, 人牙周膜干细胞, 成骨分化, Notch信号通路

Abstract: Objective: To investigate the effect of a disintegrin and metalloproteas 10 (ADAM10) on the osteogenic differentiation of human periodontal stem cells (hPDLSCs). Methods: hPDLSCs were cultured in vitro and osteogenic differentiation was induced. The mRNA expression levels of ADAM10 in the 0, 7, and 14 d of osteogenic induction were detected by RT-PCR. hPDLSCs were infected with ADAM10 overexpressed lentivirus, and the cells were divided into blank control group, no-load group, and ADAM10 overexpressed group. The levels of ADAM10 mRNA and protein expression in hPDLSCs after infection were detected by RT-PCR and Western blotting, respectively. CCK-8 was used to detect cell proliferation in each group. After osteogenic differentiation of infected hPDLSCs, alizarin red staining was performed to observe the formation of mineralized nodules in each group. The mRNA levels of osteogenic genes (ALP, Runx2, and OCN) and Notch signaling pathway related genes (Notch1, NICD, and Hes1) were detected by RT-PCR, and protein expressions of Notch signaling pathway related genes (Notch1, NICD, and Hes1) were detected by Western blot. Results: The mRNA expression level of ADAM10 decreased gradually in hPDLSCs during day 0, 7, and 14 of osteogenic differentiation (P<0.05). After infected with ADAM10 overexpressed lentivirus, the mRNA and protein expression levels of ADAM10 in hPDLSCs were significantly increased (P<0.05), and cell proliferation capacity was significantly enhanced (P<0.05). After osteogenic differentiation, compared with the control group and the no-load group, the mineralized nodule formation, mRNA expression levels of ALP, Runx2, and OCN, and protein expression levels of Notch1, NICD, and Hes1 in the ADAM10 overexpression group were significantly decreased (P<0.05). Conclusion: ADAM10 overexpression can promote the proliferation and inhibit the osteogenic differentiation of hPDLSCs, which may be related to the activation of Notch signaling pathway

Key words: a disintegrin and metalloproteas 10, human periodontal ligament stem cells, osteogenesis differentiation, Notch signaling pathway

朱永翠, 翟蕾, 闫亚姿, 李亚如. 过表达ADAM10通过调控Notch信号通路对牙周膜干细胞成骨分化的影响[J]. 口腔医学研究, 2021, 37(5): 468-473.

ZHU Yongcui, ZHAI Lei, YAN Yazi, LI Yaru. Effect of Overexpression of ADAM10 on Osteogenic Differentiation of Periodontal Ligament Stem Cells by Regulating Notch Signaling Pathway[J]. Journal of Oral Science Research, 2021, 37(5): 468-473.

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