口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (5): 500-504.DOI: 10.13701/j.cnki.kqyxyj.2018.05.010

• 口腔微生物学研究 • 上一篇    下一篇

p-Akt对牙龈卟啉单胞菌诱导的人胚胎滋养层细胞凋亡的影响

郭海盈1, 任洪芋2, 季耀庭1, 江汉1, 杜民权1*   

  1. 1. 武汉大学口腔医学院·口腔生物医学教育部重点实验室 湖北 武汉 430079;
    2. 襄阳市口腔医院 湖北 襄阳 44100
  • 收稿日期:2018-01-19 出版日期:2018-05-28 发布日期:2018-05-29
  • 通讯作者: 杜民权,E-mail: duminquan@whu.edu.cn
  • 作者简介:郭海盈(1989~ )女,河南人,硕士在读,主要从事口腔预防医学研究。
  • 基金资助:
    国家自然科学基金资助项目(编号:81371145)

Effect of p-Akt on Apoptosis of Human Extravillous Trophoblast Cells Induced by Porphyromonas Gingivalis.

GUO Hai-ying1, REN Hong-yu2, JI Yao-ting1, JIANG Han1, DU Min-quan1*   

  1. 1. MOST KLOS & KLOBM, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China;
    2. Department of Orthodontics, Xiangyang Hospital of Stomatology, Xiangyang 441000, China
  • Received:2018-01-19 Online:2018-05-28 Published:2018-05-29

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摘要： 目的:探讨牙龈卟啉单胞菌对胚胎滋养层细胞的影响及p-Akt在此过程中的作用。方法:牙龈卟啉单胞菌ATCC33277(Porphyromonas gingivalis)以感染复数(MOI)为200∶1与人胚胎滋养层细胞(HTR-8)共培养,以单纯培养人胚胎滋养层细胞为对照组;采用siRNA失活PI3K;选择倒置显微镜和共聚焦显微镜记录人胚胎滋养层细胞的形态变化;采用流式细胞术检测感染后人胚胎滋养层细胞凋亡的情况;用Real Time-PCR检测PI3K siRNA敲低的效果。结果:牙龈卟啉单胞菌感染人胚胎滋养层细胞12 h时,细胞的凋亡率与正常组相比无统计学意义;感染24 h时,细胞凋亡明显增多(P<0.01);感染48 h时,细胞凋亡与对照组相比更为明显(P<0.001)。siRNA预先处理HTR-8细胞再与牙龈卟啉单胞菌共培养48 h后,siRNA处理组的PI3K mRNA的相对量明显低于干扰组(P<0.01);同时,细胞凋亡率>45%,明显高于单纯感染牙龈卟啉单胞菌组(P<0.01),而干扰组的细胞凋亡率与单纯刺激组相比无统计学差异。结论:牙龈卟啉单胞菌能够诱导胚胎滋养层细胞发生凋亡。在此过程中,p-Akt被激活,并对凋亡有一定的抑制作用。提示,牙周炎促使早产低体重儿的娩出与牙龈卟啉单胞菌诱导胚胎滋养层细胞凋亡有关。

关键词: 牙周炎, 牙龈卟啉单胞菌, 早产低体重儿, 凋亡

Abstract: Objective: To study the effect of Porphyromonas gingivalis on human extravillous trophoblast cell line (HTR-8) and the role of p-Akt in this process. Methods: HTR-8 cells were infected with Porphyromonas gingivalis ATCC 33277 at a multiplicity of infection (MOI) of 200 or 0. The target siRNA was chosen to inactivate PI3K, inverted microscope and confocal microscope were used to record the morphological changes of HTR-8 cells, flow cytometry was used to detect the apoptosis of HTR-8 cells after infection with Porphyromonas gingivalis, and real time-PCR was used to test the inactivation of siRNA whose target was PI3K. Results: When HTR-8 cells were infected with Porphyromonas gingivalis for 12 hours, there was no statistical significance of the apoptosis rate in MOI 200 group compared with MOI 0 group. HTR-8 cells were infected for 24 hours, the number of apoptotic cells in MOI 200 group significantly increased (P<0.01). Infected for 48 hours, the extent of apoptosis was more obvious compared with control group MOI 0 group (P<0.001). When HTR-8 cells were pretreated with siRNA and then co-cultured with Porphyromonas gingivalis for 48 hours (SI 200 Group), the relative amounts of PI3K mRNA in SI 200 group was much lower than that in interference group (C 200 group) (P<0.01). At the same time, the apoptosis rate of SI 200 group was greater than 45%, which was significantly higher than that of MOI 200 group (P<0.01); and there was no statistical difference of apoptosis rate between MOI 200 group and C 200 group. Conclusion: Porphyromonas gingivalis does induce apoptosis in human extravillous trophoblast cells and p-Akt is activated to inhibit the apoptosis of HTR-8 cells to some extent in this process. All of these suggest that the mechanism by which periodontitis promotes preterm low birth weight is that Porphyromonas gingivalis induced human extravillous trophoblastic cells apoptosis.

Key words: Periodontitis, Porphyromonas gingivalis, Preterm low birth weight, Apoptosis