›› 2019, Vol. 10 ›› Issue (4): 174-179.

• 论著 • 上一篇    下一篇

微小RNA在慢性咀嚼肌疼痛中作用的初步探索

张文静1,孟贺2,谢秋菲3   

  1. 1. 北京大学口腔医学院 · 口腔医院修复科
    2. 深圳大学总医院口腔科
    3. 北京大学口腔医学院?口腔医院修复科
  • 收稿日期:2019-09-14 修回日期:2019-12-04 出版日期:2019-12-25 发布日期:2020-01-06
  • 通讯作者: 谢秋菲 E-mail:xieqiuf@163.com
  • 基金资助:
    国家自然科学基金

Preliminary exploration of the role of microRNA in chronic masticatory myalgia

  • Received:2019-09-14 Revised:2019-12-04 Online:2019-12-25 Published:2020-01-06

摘要: 目的:研究微小RNA(miRNA)在慢性咀嚼肌疼痛(CMM)中的调控作用。方法:收集北京大学口腔医院颞下颌关 节病及口颌面痛门诊CMM患者23名,以性别及年龄匹配的健康志愿者作为对照,静脉采血2mL。课题组前期实验利用Targetscan 数据库和DIANA数据库初步筛选出可能通过丝裂原活化蛋白激酶激酶3(MEK3)基因调控CMM的miR-15a-5p和miR-19b-3p 作为目标miRNA,用qRT-PCR进一步检测可能相关的miRNA,进行细胞转染和miRNA过表达实验,并通过qRT-PCR及Western blot验证miRNA与MEK3基因的关系。结果:qRT-PCR检测发现miR-19b-3p在CMM患者组显著下调(P<0.05),miR-15a-5p在 两组间无显著性差异(P>0.05),后续实验选择miR-19b-3p进一步验证miRNA与MEK3基因的关系。结果发现在人单核细胞白 血病细胞(THP-1)中,miR-19b-3p过表达组的MEK3基因的mRNA和蛋白表达在24h、48h和72h时与miR-NC组相比均无显著 性差异(P>0.05)。结论:miR-19b-3p可能参与了CMM,但可能不是通过与MEK3基因直接结合发挥作用。

关键词: miR-19b-3p, MEK3, 慢性咀嚼肌疼痛

Abstract: Objective: To investigate the regulatory effect of miRNA on chronic masticatory myalgia(CMM). Methods: Twenty-three CMM patients were recruited from the Center for TMD and Orofacial Pain of Peking University School and Hospital of Stomatology, 23 gender- and age- matched healthy people recruited as control group. A total of 2 mL peripheral blood was collected from each participant of two groups. MiR-15a-5p and miR-19b-3p were preliminarily screened out as target miRNAs which are associated with CMM by regulating MEK3 according to Targetscan database and DIANA database in the previous experiments. QRT-PCR was used to select the possible miRNA further. Subsequently, the cell transfection and miRNA-overexpression experiment were conducted to verify the relationship of miRNA and MEK3 gene with qRT-PCR and Western blot. Results: MiR-19b-3p expressions were significantly decreased in CMM group compared to healthy control group using qRT-PCR assays (P<0.05), and miR-15a-5p expressions were not significantly different between these two groups(P>0.05). MiR-19b-3p was selected to verify the relationship between miRNA and MEK3 gene in the following experiments. The results showed that the mRNA and protein of MEK3 gene had no significant difference between miR-19b-3p-overexpression group and miR-NC group in human leukemia monocytic cell line (THP-1 cell) at 24h、48h and 72h ( P>0.05). Conclusions: MiR-19b-3p may be involved in CMM and its mechanism is likely to be complicated. It may not work by binding to the MEK3 gene directly.

Key words: miR-19b-3p, MEK3, chronic masticatory myalgia