摘要
目的:验证TRPM4(transient receptor potential melastatin-4)在口腔颊黏膜成纤维细胞的表达,探讨其对人口腔颊黏膜成纤维细胞迁移与增殖中的作用。方法:收集人正常颊黏膜组织进行原代细胞提取与原代培养;采用RT-PCR和细胞免疫荧光方法检测成纤维细胞中TRPM4的表达。全细胞膜片钳记录成纤维细胞TRPM4通道的全细胞电流。MTT法和细胞划痕实验分别检测应用TRPM4通道特异性阻断剂(9-菲酚)或特异性siRNA抑制TRPM4通道的功能对成纤维细胞增殖和迁移的影响。结果:人口腔颊黏膜成纤维细胞功能性表达TRPM4。抑制TRPM4通道的功能能够明显降低成纤维细胞的增殖和迁移能力。结论:TRPM4通道参与成纤维细胞增殖以及迁移能力的调控。
Objective:To verify the expression of TRPM4 in buccal mucosal fibroblasts and explore its effects on migration and proliferation of buccal fibroblasts.Methods:Human buccal mucosa fibroblasts were isolated and cultured in vitro,and the expression of TRPM4 in fibroblasts was evaluated by reverse transcription polymerase chain reaction(RT-PCR)and immunofluorescence.Whole cell patch clamp were performed to record the whole cell currents of TRPM4 channels.The effects of 9-phenanthrene or TRPM4 specific siRNA on the proliferation and migration of fibroblasts were estimated by MTT assay and cell scratch test,respectively.Results:TRPM4 was functional expressed in human buccal mucosa fibroblasts.Suppression the function of TRPM7 channel by 9-phenanthrene or specific siRNA inhibited the proliferation and migration capability of the cells.Conclusion:TRPM4 channels is involved in the regulation of proliferation and migration in human buccal mucosa fibroblasts.
作者
王薛藤
聂敏海
左东川
邓浩
曾锦
WANG Xueteng;NIE Minhai;ZUO Dongchuan;DENG Hao;ZENG Jin(Department of Periodontal Mucosa, Affiliated Stomatological Hospital, Southwestern Medical University, Luzhou 646000, China;Oralfacial Reconstruction and Regeneration Laboratory, Luzhou 646000, China;Cardiovascular Research Institute, Southwest Medical University, Luzhou 646000, China;Department of Orthodontics, Affiliated Stomatological Hospital, Southwestern Medical University, Luzhou 646000, China)
出处
《口腔医学研究》
CAS
北大核心
2020年第5期490-495,共6页
Journal of Oral Science Research
基金
四川省科技厅课题(川科技[2018]26号,项目编号2018JY0401)
西南医科大学市-校-附属口腔医院联合项目(编号:0800103009)
西南医科大学应用基础研究计划项目(编号:2017-ZRZD-001)。
