口腔医学研究

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (10): 1052-1055.DOI: 10.13701/j.cnki.kqyxyj.2017.10.008

• 基础研究论著 • 上一篇    下一篇

NF-κB非经典途径IKKα调控的Maspin在人牙周膜细胞中的表达

王锦华，张芳*，李霞，陈庆勇   

  1. 山西医科大学口腔医学系 山西 太原 030001
  • 收稿日期:2017-04-10 出版日期:2017-10-20 发布日期:2017-10-24
  • 通讯作者: 张芳,Email:zhangfangdoctor@163.com
  • 作者简介:王锦华(1989~ ),女,陕西渭南人,医师,硕士在读,主要从事牙周病学研究。
  • 基金资助:
    山西省科技攻关项目(编号:20150313010-3)高等学校博士启动资金(编号:03201321)

Preliminary Study on Expression of IKKα-regulated Maspin in Human Periodontal Ligament Cells

ANG Jin-hua, ZHANG Fang*, LI Xia, CHEN Qing-yong   

  1. Department of Stomatology, Shanxi Medical University, Taiyuan 030001, China.
  • Received:2017-04-10 Online:2017-10-20 Published:2017-10-24

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摘要： 目的:观察NF-κB非经典通路IKKα调控的Maspin在HPDLCs中的表达。方法:组织块法原代培养HPDLCs。倒置显微镜及SP免疫组化鉴定细胞来源。免疫细胞化学定位Maspin在HPDLCs中的表达。实验分空白对照组(NC)、LPS1-4组(LPS终浓度为0.1、1、10、50 mg/L)。RT-PCR及Western Blot分别检测LPS刺激下HPDLCs中Maspin、IKKα mRNA及蛋白的表达。Elisa检测炎性因子TNF-α和IL-1β的分泌。结果:Maspin在正常与LPS刺激的HPDLCs中均呈胞浆表达。随着LPS浓度增加 Maspin蛋白表达下调,LPS1-4与NC组相比 (P<0.05);IKKα蛋白表达上调,LPS1-4与NC组相比 (P<0.05)。mRNA表达与蛋白表达趋势相同。同时,二者相关性分析可知,Maspin与IKKα mRNA及蛋白表达呈负相关。Elisa结果显示TNF-α、IL-1β随着LPS浓度增加分泌增加,且与Maspin蛋白表达呈负相关。结论:Maspin在HPDLCs中呈胞浆表达,LPS可介导HPDLCs中Maspin表达下调,与TNF-α、IL-1β呈负相关,且Maspin 表达下调可能经由NF-κB非经典途径IKKα调控。

关键词: 牙周炎, 乳腺丝氨酸蛋白酶抑制剂, 牙周膜细胞, IκB激酶

Abstract: Objective: To observe the expressions of IKKα-regulated Maspin in NF-κB non-classical pathway in human periodontal ligament cells (HPDLCs). Methods: HPDLCs were carried out using tissue culture method and identified by inverted microscope and SP immunohistochemistry. Cells were divided into blank control group (NC) and LPS1-4 groups (LPS: 0.1,1,10,50 mg/L). Protein and mRNA levels of Maspin and IKKα were measured by Western Blot and RT-PCR respectively, and the secretions of TNF-α and IL-1β were tested using Elisa assay. Results: Maspin in HPDLCs was expressed in cytoplasm. Western blot showed that with the increase of LPS concentration, the expression of Maspin protein was down regulated and IKKα protein was up-regulated compared with NC group (P<0.05). The expression of mRNA was similar to that of protein expression. Secretions of TNF-α and IL-1β were enhanced accompanied with increasing concentrations of LPS, and negatively correlated with Maspin. Conclusion: The expression of Maspin in HPDLCs was down-regulated by LPS and showed a negative correlation with TNF-α and IL-1β, and may be regulated by NF-κB non-classical pathway IKKα.

Key words: Periodontitis, Maspin , HPDLCs, IKKα

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