长链非编码RNA AWPPH通过调控Notch信号通路对人牙周膜细胞增殖和成骨向分化的影响  

Effect of lnc RNA AWPPH on human periodontal ligament cell proliferation and osteogenic differentiation by regulating Notch signaling pathway

作　　者：迪丽达尔·塔西甫拉提 牛雅琪 热依沙·阿布都克依木 王玲[1] Dilidaer·Taxifulati;NIU Ya-qi;Reyisha·Abudukeyimu;WANG Lin(Department of Oral Surgery Clinic,The First Affiliated Hospital of Xinjiang Medical University(Affiliated Hospital of Stomatology),Xin-jiang Uygur Autonomous Region Institute of Stomatology.Wulumuqi 830001,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)口腔外科门诊,新疆维吾尔自治区口腔医学研究所,新疆乌鲁木齐830001

出　　处：《上海口腔医学》2023年第1期23-27,共5页Shanghai Journal of Stomatology

基　　金：新疆维吾尔自治区自然科学基金(2016D01C250)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)AWPPH通过调控Notch信号通路对人牙周膜细胞增殖和成骨向分化的影响及分子机制。方法:体外培养人牙周膜细胞,并进行成骨分化诱导,使用定量即时聚合酶链锁反应(qRT-PCR)实验检测第0、3、7、14天细胞AWPPH表达水平。将人牙周膜细胞分为空白对照组(NC)、空载体组(vector)、AWPPH过表达组(AWPPH)和过表达AWPPH+通路抑制剂组(AWPPH+DAPT)。qRT-PCR实验检测AWPPH表达水平;噻唑蓝(MTT)、克隆实验检测细胞增殖情况。蛋白质免疫印迹(Western blot)实验检测碱性磷酸酶(ALP)、骨桥蛋白(OPN)、骨钙素(OCN)、Notch1和Hes1蛋白表达。采用SPSS 21.0软件包对数据进行统计学分析。结果:牙周膜细胞成骨分化第0、3、7、14天,AWPPH表达水平逐渐降低。过表达AWPPH可增加牙周膜细胞吸光度(A)值、克隆细胞数目,上调ALP、OPN、OCN、Notch1和Hes1蛋白表达。加入通路抑制剂DAPT后,细胞A值、克隆细胞数目降低,Notch1、Hes1、ALP、OPN和OCN蛋白表达降低。结论:过表达AWPPH可能通过降低Notch信号通路相关蛋白表达,抑制牙周膜细胞增殖和成骨分化。PURPOSE: To explore the effect and molecular mechanism of long non-coding RNA(lncRNA) AWPPH on proliferation and osteogenic differentiation of human periodontal ligament cells by regulating the Notch signaling pathway.METHODS: Human periodontal ligament cells were cultured in vitro, and osteogenic differentiation was induced. Quantitative real-time polymerase chain reaction(qRT-PCR) experiment were used to detect the AWPPH expression level of cells at 0, 3, 7, and 14 days. Human periodontal ligament cells were divided into blank control group(NC), empty vector group(vector), AWPPH overexpression group(AWPPH), and overexpression AWPPH+ pathway inhibitor group(AWPPH+DAPT). qRT-PCR experiment was used to detect the expression level of AWPPH;thiazole blue(MTT), cloning experiment was used to detect cell proliferation. Western blot was performed to detect the protein expression of alkaline phosphatase(ALP), osteopontin(OPN), osteocalcin(OCN), Notch1 and Hes1. SPSS 21.0 software package was used for statistical anal-ysis. RESULTS: The AWPPH expression level in periodontal ligament cells decreased after 0, 3, 7, and 14 days of osteogenic differentiation. Overexpression of AWPPH increased the A value of periodontal ligament cells, the number of cloned cells, and up-regulated the protein expression of ALP, OPN, OCN, Notch1, and Hes1. After adding the pathway inhibitor DAPT, the A value and the number of cloned cells decreased, and the protein expression of Notch1, Hes1, ALP,OPN, and OCN decreased. CONCLUSIONS: Overexpression of AWPPH may inhibit the proliferation and osteogenic differentiation of periodontal ligament cells by reducing the expression of related proteins in the Notch signaling pathway.

关 键 词：AWPPH NOTCH信号通路 人牙周膜细胞 增殖 成骨向分化 

分 类 号：R781.4[医药卫生—口腔医学]