Nell-1蛋白通过Wnt/β-catenin通路调控人上颌窦黏膜来源间充质干细胞成骨分化  

Nell-1 Regulates Osteogenic Differentiation of Human Maxillary Sinus Membrane-derived Mesenchymal Stem Cells through Wnt/β-catenin Pathway.

作　　者：徐静 吕慧欣 王一涵 鲍鑫 张逸 周延民[1] XU Jing;LV Huixin;WANG Yihan;BAO Xin;ZHANG Yi;ZHOU Yanmin(Department of Implantology,Hospital of Stomatology,Jilin University,Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling,Changchun 130012,China;Department of Prosthodontics,Hospital of Stomatology,Jilin University,Changchun 130012,China)

机构地区：[1]吉林大学口腔医院种植科,吉林省牙发育及颌骨重塑与再生重点实验室,吉林长春130012 [2]吉林大学口腔医院修复科,吉林长春130012

出　　处：《口腔医学研究》2023年第3期227-235,共9页Journal of Oral Science Research

基　　金：国家自然科学基金面上项目(编号:82071152)。

摘　　要：目的:研究不同浓度Nell-1(Nel-like type 1 molecule)蛋白对人上颌窦黏膜来源间充质干细胞(maxillary sinus membrane-derived mesenchymal stem cells,MSMSCs)成骨分化的影响。方法:体外分离培养并鉴定人MSMSCs。使用Nell-1蛋白(10、100、1000 ng/mL)诱导MSMSCs,利用CCK-8法和细胞活死染色法检测细胞活性变化;碱性磷酸酶活性检测细胞成骨能力改变;实时定量荧光聚合酶链反应(qRT-PCR)检测成骨相关因子mRNA表达情况;免疫荧光染色和Western blot法检测成骨蛋白表达情况。结果:Nell-1蛋白对MSMSCs有一定的促增殖作用;碱性磷酸酶活性检测显示各实验组成骨效果优于对照组;qRT-PCR、免疫荧光染色和Western blot结果显示,100 ng/mL的Nell-1显著促进β-catenin、Runx-2、OCN等mRNA及蛋白的表达(P<0.01)。结论:Nell-1蛋白可通过上调Wnt/β-catenin信号通路的表达提高MSMSCs体外成骨分化能力。Objective:To investigate the effect of Nell-1 protein on osteogenic differentiation of human maxillary sinus membrane-derived mesenchymal stem cells(MSMSCs).Methods:The MSMSCs were induced by Nell-1 protein(10,100,1000 ng/mL),and cell activity changes were detected using CCK-8 assay and cell live-dead staining.The changes in osteogenic ability were detected by alkaline phosphatase activity.mRNA expression of osteogenic-related factors was detected by quantitative real-time fluorescence PCR(qRT-PCR).Immunofluorescence staining and western blot were used to detect osteogenic protein expression.Results:Nell-1 protein had a promote effect on proliferative of MSMSCs.Alkaline phosphatase activity assay showed that the osteogenic effect of each experimental group was better than that of the control group.qRT-PCR,immunofluorescence staining,and western blot showed that 100 ng/mL of Nell-1 significantly promoted the mRNAs and protein expression ofβ-catenin,Runx-2,OCN,and others(P<0.01).Conclusion:Nell-1 protein can improve the osteogenic differentiation ability of MSMSCs in vitro by upregulating the expression of Wnt/β-catenin signaling pathway.

关 键 词：间充质干细胞 骨再生 上颌窦 Nell-1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]