NF-κB非经典途径IKKα调控的Maspin在人牙周膜细胞中的表达

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

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)、LPS_(1-4)组(LPS终浓度为0.1、1、10、50mg/L)。RT-PCR及Western Blot分别检测LPS刺激下HPDLCs中Maspin、IKKαmRNA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,LPS_(1-4)与NC组相比(P<0.05);IKKα蛋白表达上调,LPS_(1-4)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IKKαmRNA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。

Objective： To observe the expressions of IKKα- regulated Maspin in NF-κβ 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. Ceils 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 IKKa were meas- ured 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 IKKa 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-βB non--classical pathway IKKα.