miR-210促进大鼠牙髓干细胞的增殖和牙源性分化  

Micro RNA-210 promotes the proliferation and odontogenic differentiation of rat dental pulp stem cells

作　　者：王艳玲 左春然 王静 杨琨 王守儒 张小平 Wang Yanling;Zuo Chunran;Wang Jing;Yang Kun;Wang Shouru;Zhang Xiaoping(Department of Stomatology, Henan Traditional ChineseMedicine Hospital (Second Affiliated Hospital of Henan University of Chinese Medicine), Zhengzhou 450002, Henan Province, China)

机构地区：[1]河南省中医院(河南中医药大学第二附属医院)口腔科

出　　处：《中国组织工程研究》2019年第25期4004-4010,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省科技厅省部级课题(172102310613),项目负责人:王艳玲~~

摘　　要：背景:miR NA表达谱预测miR-210可能在牙髓干细胞向牙本质和成骨方向分化中发挥作用,但具体作用尚不明确。目的:探讨miR-210在大鼠牙髓干细胞增殖和牙源性分化中的作用。方法:经河南中医药大学第二附属医院伦理委员会批准,收集5只SD大鼠牙髓组织,分离获得牙髓干细胞并分为5组:正常对照组、miR-210模拟物阴性对照组、miR-210模拟物组、miR-210抑制物阴性对照组和miR-210抑制物组。实时荧光定量PCR和Western blot检测细胞中miR-210、ALP、DSPP、DMP-1、GIT2、OPNmR NA的表达和DSPP、DMP-1、OPN和OCN蛋白的表达,CCK-8细胞增殖检测试剂盒检测细胞的增殖能力,比色法检测细胞碱性磷酸酶活性,茜素红染色检测矿物质合成能力。结果与结论:(1)miR-210模拟物组细胞的增殖活力高于其他4组,差异有显著性意义(P <0.01);miR-210抑制物组细胞的增殖活力低于其他4组,差异有显著性意义(P <0.01);(2)miR-210模拟物组细胞的碱性磷酸酶活性高于其他4组,差异有显著性意义(P <0.01);miR-210抑制物组细胞的碱性磷酸酶活性低于其他4组,差异有显著性意义(P <0.01);(3)茜素红染色结果显示,miR-210模拟物组细胞表面布满钙盐沉积形成的大小不等的暗红色结节,数量多于其他4组,miR-210抑制物组细胞表面的暗红色钙结节散在分布,数量少于其他4组;(4)miR-210模拟物组细胞中miR-210、ALP、DSPP、DMP1、GIT2、OPN mRNA和DSPP、DMP1、OPN、OCN蛋白的表达高于其他4组,差异有显著性意义(P <0.01);miR-210抑制物组细胞中miR-210、ALP、DSPP、DMP1、GIT2、OPN mRNA和DSPP、DMP1、OPN、OCN蛋白的表达低于其他4组,差异有显著性意义(P <0.01);(5)结果提示,miR-210能够促进大鼠牙髓干细胞增殖与牙源性分化。BACKGROUND: MicroRNAs profile predicts that microRNAs-210 may play a role in the differentiation of dental pulp stem cells into dentin and osteogenesis, but the specific role remains unclear.OBJECTIVE: To investigate the role of microRNA-210 in the proliferation and odontogenic differentiation of rat dental pulp stem cells.METHODS: The study was approved by the Ethics Committee of the Second Affiliated Hospital of Henan University of Chinese Medicine,China. The dental pulp tissues were collected from the mandibles of five Sprague-Dawley rats, and the pulp stem cells were isolated and identified. The dental pulp stem cells were divided into five groups. The normal control group was not treated. The cells in the miR-210 mimic negative control group were transfected with the miR-210 mimic negative control. The cells in the miR-210 mimic group were transfected with the miR-210 mimic. The cells in the miR-210 inhibitor negative control group were transfected with the miR-210 inhibitor negative control. The cells in the miR-210 inhibitor group were transfected with the miR-210 inhibitor negative control. Real-time quantitative PCR was used to detect the expression of microRNA-210, alkaline phosphatase, dentin sialophosphoprotein, osteopontin, osteocalcin, dentin matrix phosphoprotein-1 and GPCR-kinase interacting protein-2 in cells. Western blot was used to detect the expression of dentin sialophosphoprotein, osteopontin, osteocalcin and dentin matrix phosphoprotein-1 in cells. Cell counting kit-8 was used to detect cell proliferation. Cell alkaline phosphatase activity was detected by colorimetry. Alizarin red staining was used to detect the ability of mineral synthesis.RESULTS AND CONCLUSION:(1) The proliferation activity of miR-210 mimic group was significantly higher than that of the other four groups(P < 0.01), while the proliferation activity of miR-210 inhibitor group was significantly lower than that of the other four groups(P < 0.01).(2) The activity of alkaline phosphatase in the miR-210 mimic group was signi

关 键 词：牙髓干细胞 MIRNA MIR-210 牙源性分化 细胞增殖 碱性磷酸酶活性 矿物质合成能力 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]