糖基化终末产物刺激下长链非编码RNA H19通过WNT/β-catenin信号通路调控MC3T3细胞骨向分化  

Long non-coding RNA H19 regulates the osteogenic differentiation of MC3T3-E1 cells through WNT/β-catenin signaling pathway under the stimulation of AGEs

作　　者：邓超 强诗雨 马一凡 阮晓旭 孙翼 李雪雪 DENG Chao;QIANG Shiyu;MA Yifan;RUAN Xiaoxu;SUN Yi;LI Xuexue(Anhui Province Engineering Research Center for Dental Materials and Application,School of Stomatology,Wannan Medical College,,Wuhu 241002,China)

机构地区：[1]安徽省口腔材料与应用转化工程研究中心,皖南医学院口腔医学院,芜湖241002

出　　处：《实用口腔医学杂志》2024年第6期765-769,共5页Journal of Practical Stomatology

基　　金：安徽省教育厅高校优秀拔尖人才培育项目(编号:gxyq-2020024);皖南医学院青年国内访学研修资助项目(编号:wyqnyx202008);安徽省高校自然科学基金项目(编号:KJ2021A0854);国家级大学生创新创业项目(编号:202010368034);芜湖市科技项目(编号:2022jc35);皖南医学院校重点项目科研基金(编号:WK2021Z04)。

摘　　要：目的:探究长链非编码RNA H19(LncRNA H19)在糖基化终末产物(AGEs)刺激下对小鼠前成骨细胞MC3T3-E1成骨分化的调控作用及其可能的机制。方法:10μg/mL AGEs浓度刺激下培养MC3T3-E1细胞,7 d后碱性磷酸酶染色、21 d后茜素红染色观察细胞矿化情况;RT-PCR检测细胞中ALP、Runx-2、OCN、SP7以及LncRNA H19 mRNA的表达水平,Western blotting检测细胞中Runx-2蛋白表达水平;采用细胞转染技术过表达LncRNA H19,RT-PCR检测LncRNA H19的转染效率;并检测转染LncRNA H19后细胞中骨基因的表达水平、ALP染色情况以及β-catenin的表达情况。结果:AGEs刺激下MC3T3-E1细胞碱性磷酸酶染色变浅、钙化结节形成减少,ALP、Runx-2、OCN、SP7、LncRNA H19基因表达水平降低(P<0.05),Runx-2蛋白水平表达下降;转染过表达LncRNA H19(lv-H19)后,LncRNA H19表达水平显著上调(P<0.05),ALP、Runx-2、OCN、SP7基因表达水平上升(P<0.05),碱性磷酸酶染色变深,β-catenin的表达增加。结论:AGEs刺激下,LncRNA H19可能通过WNT/β-catenin的表达影响了MC3T3-E1细胞的骨向分化。Objective:To explore the regulatory effects of long non-coding RNA H19(LncRNA H19)on the osteogenic differentiation of MC3T3-E1 cells under the stimulation of advanced glycation end products(AGEs).Methods:MC3T3-E1 cells were cultured under the stimulation of 10μg/mL AGEs.The cells were observed by alkaline phosphatase(ALP)staining after 7 d and alizarin red staining after 21 d culture respectively.Cell transfection technology was used to overexpress LncRNA H19 in the cells,RT-qPCR was used to detect the mRNA expression levels of ALP,Runx-2,OCN,SP7 before and after transfection of LncRNA H19.Western blotting was used to detect the protein expression of Runx-2 andβ-catenin in the cells.Results:Under the stimulation of AGEs,the ALP staining color of MC3T3-E1 cells became lighter,the formation of calcified nodules was reduced,the mRNA expression levels of ALP,Runx-2,OCN,SP7 and LncRNA H19 were decreased(P<0.05),and the protein expression level of Runx-2 was decreased.After transfection of LncRNA H19(lv-H19),the mRNA expression of LncRNA H19 was significantly up-regulated(P<0.05),the mRNA expression of ALP,Runx-2,OCN and SP7 genes was increased(P<0.05),ALP staining was deeper,and the expression ofβ-catenin was increased.Conclusion:Under the stimulation of AGEs,LncRNA H19 may affect the osteogenic differentiation of MC3T3-E1 cells by activating the expression of WNT/β-catenin.

关 键 词：糖基化终末产物 长链非编码RNA H19 小鼠前成骨细胞 骨向分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]