口腔医学研究

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (5): 477-480.DOI: 10.13701/j.cnki.kqyxyj.2020.05.018

• 牙周病学研究 • 上一篇    下一篇

脂多糖抑制牙周膜细胞骨唾液酸蛋白表达中微小RNA的筛选与验证

张源, 李新月*   

  1. 天津市口腔医院牙周病科 天津 300041
  • 收稿日期:2019-09-09 出版日期:2020-06-16 发布日期:2020-06-18
  • 通讯作者: * 李新月,E-mail: lisaxinyue1215@126.com
  • 作者简介:张源(1989~ ),女,江苏人,主治医师,硕士,主要从事牙周再生的研究。

Screening and Identification of microRNA on BSP Gene of Human Periodontal Ligament Cells Depressed by Lipopolysaccharide

ZHANG Yuan, LI Xinyue*   

  1. Department of Periodontolgoy Tianjin Stomatological Hospital, Tianjin 300041, China
  • Received:2019-09-09 Online:2020-06-16 Published:2020-06-18

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摘要： 目的:筛选和鉴定靶向调节骨唾液酸蛋白(bone sialoprotein,BSP)基因的微小 RNA( microRNA,miRNA),研究其在高浓度牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g.)脂多糖(lipopolysaccharide,LPS)抑制人牙周膜细胞(human periodontal ligament cells,hPDLCs)BSP基因表达中的靶向调节作用。方法:通过生物信息学预测和双荧光素酶报告基因分析系统筛选并验证能够靶向调节BSP基因的微小 RNA(microRNA,miRNA);获得miR-129过表达和表达抑制的载体质粒,以此转染hPDLCs,10 mg/L P.g. LPS 处理各组细胞8 h后,采用qRT-PCR和Western blotting检测BSP基因和蛋白的表达水平。结果:10 mg/L P.g. LPS 处理hPDLCs 8 h后,miR-129相对表达量增加,而BSP mRNA相对表达量降低。P.g. LPS处理转染miR-129 mimics的hPDLCs后,BSP mRNA相对表达量低于对照组,转染miR-129 inhibitor的hPDLCs经P.g. LPS处理后BSP mRNA相对表达量高于对照组(P＜0.05)。结论:在P.g. LPS影响hPDLCs成骨活性过程中,miR-129可能是抑制BSP基因表达的中间环节之一。

关键词: 微小RNA, 牙龈卟啉单胞菌, 脂多糖, 骨唾液酸蛋白, 骨吸收

Abstract: Objective: To screen and identify miRNA which regulate gene expression of BSP and to analyze miRNA to regulate BSP gene expression during the osteogenic differentiation of hPDLCs depressed by LPS from P.g. Methods: Bioinformatics prediction and dual luciferase reporter gene analysis system were used to screen and verify the miR-129 that could be targeted to regulate BSP gene. miR-129 overexpression and expression inhibition vector plasmids were obtained to transfect hPDLCs. After treated each group of cells with 10mg/L P.g. LPS for 8 hours, the expression level of BSP was detected by qRT-PCR and western blotting. Results: Treatments with 10mg/L of P.g. LPS for 8 hours up-regulated the expression of miR-129, whereas BSP mRNA expression was induced. The expression levels of BSP in the experimental groups were lower than those in the mimic control group when miR-129 expression increased (P＜0.05). The protein expression levels of BSP in the experimental groups were higher than those in the inhibitor control group when miR-129 expression decreased(P＜0.05). Conclusion: In the process of P.g. LPS affecting osteogenic activity of hPDLCs, miR-129 may reduce its osteogenic activity by inhibiting BSP.

Key words: microRNA, porphyromonas gingivalis, lipopolysaccharide, bone sialoprotein, bone destruction