可溶性环氧化物水解酶抑制剂对暴露于HEMA的人牙髓干细胞细胞迁移及成牙分化的影响  

Effect of Soluble Epoxide Hydrolase Inhibitor on Migration and Odontogenic Differentiation of Human Dental Pulp Stem Cells Exposed to HEMA

作　　者：罗加欣 黄旌 王渝灵 冯燕 党海霞 LUO Jiaxin;HUANG Jing;WANG Yuling;FENG Yan;DANG Haixia(Oral&Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory,The Affiliated Stomatological Hospital of Southwest Medical University,Luzhou 646000,China.)

机构地区：[1]西南医科大学附属口腔医院口颌面修复重建和再生泸州市重点实验室,四川泸州646000

出　　处：《口腔医学研究》2024年第2期166-171,共6页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81771032);西南医科大学青年基金(编号:00031520);西南医科大学口腔医院一般项目(编号:201906)。

摘　　要：目的:探讨在树脂单体甲基丙烯酸羟乙酯(2-hydroxyethyl methacrylate, HEMA)暴露环境下,可溶性环氧化物水解酶抑制剂(TPPU)对人牙髓干细胞(human dental pulp stem cells, hDPSCs)迁移及成牙分化的影响。方法:分离提取hDPSCs,并利用茜素红染色和流式细胞仪鉴定其干性。以无添加药物的诱导培养基为对照组,以HEMA、HEMA+TPPU为实验组,利用CCK-8检测细胞活性;利用细胞划痕实验检测细胞迁移;利用碱性磷酸酶染色法进行染色,并检测碱性磷酸酶(ALP)活性;利用实时荧光定量聚合酶链反应(RT-qPCR)检测成牙分化相关基因Runt相关转录因子2(Runx2)、牙本质基质蛋白-1(DMP-1)和牙本质涎磷蛋白(DSPP)的相对表达;利用茜素红染色检测矿化结节形成情况。结果:HEMA组细胞活性较对照组显著降低(P<0.05),HEMA+TPPU组较HEMA组增加(P<0.05)。HEMA组细胞迁移率较对照组显著降低(P<0.05),HEMA+TPPU组较HEMA组增加(P<0.05)。在成牙诱导分化过程中,用2 mmol/L的HEMA处理细胞后,ALP活性较对照组降低(P<0.05),相关成牙分化基因表达也显著降低(P<0.05)。而HEMA+TPPU组ALP活性较HEMA组显著升高(P<0.05),且基因Runx2、DMP-1、DSPP表达也显著增加(P<0.05)。此外茜素红染色结果也显示HEMA+TPPU组和对照组的相对钙含量均较HEMA组显著升高(P<0.05)。结论:HEMA可在一定程度上抑制hDPSCs的细胞迁移和成牙分化能力,而TPPU可部分逆转HEMA对细胞迁移和成牙分化能力的抑制作用。Objective:To investigate the effect of soluble epoxide hydrolase inhibitor(TPPU)on the migration and odontogenic differentiation of human dental pulp stem cells(hDPSCs)exposed to 2-hydroxyethyl methacrylate(HEMA).Methods:hDPSCs were isolated and extracted,and then identified by alizarin red staining and flow cytometry.hDPSCs without drug culture were used as the control group,and HEMA and HEMA+TPPU were used as the experimental group.hDPSCs activity was detected by CCK-8.hDPSCs migration was detected by Scratch test.ALP activity was detected and be quantified by alkaline phosphatase staining and quantitative test kit.Realtime quantitative fluorescent PCR(RT-qPCR)was used to detect the relative expression of Runx2,DMP-1,and DSPP related to odontogenic differentiation.Alizarin red staining was used to detect the formation of mineralized nodules.Results:The cell activity of HEMA group was lower than that of control group(P<0.05),and the cell activity of HEMA+TPPU group was higher than that of HEMA group(P<0.05).The cell mobility of HEMA group was significantly decreased compared with that of control group(P<0.05),and the cell mobility of HEMA+TPPU group was significantly increased compared with that of HEMA group(P<0.05).After the cells were treated with 2 mmol/L HEMA in the process of odontogenic differentiation,alkaline phosphatase activity was decreased when compared with control group(P<0.05),and the expression of odontogenic differentiation related genes was significantly decreased(P<0.05).The activity of alkaline phosphatase in the HEMA+TPPU group was significantly higher than that in HEMA group (P <0.05), and the expressions of Runx2, DMP-1, and DSPP were also significantly increased (P <0.05). In addition, alizarin red staining showed that the relative calcium contents of HEMA+TPPU group and control group were significantly higher than that of HEMA group (P <0.05). Conclusion: HEMA can inhibit the cell migration and odontogenic differentiation of hDPSCs to some extent, and TPPU can partially reverse the inhi

关 键 词：可溶性环氧化物水解酶抑制剂 HEMA 牙髓干细胞 细胞迁移 成牙分化 

分 类 号：R781.1[医药卫生—口腔医学]