牙龈成纤维细胞表达蛋白酶激活受体的类型及作用研究  

Expressions of protease-activated receptors in human gingival fibroblasts and its functions in periodontitis

作　　者：张迪亚[1] 胡玲静[2] 李盛来[3] 盛列平[1] 曹筝[1] 陈莉丽[2] 

机构地区：[1]浙江大学医学院附属邵逸夫医院牙科,杭州310016 [2]浙江大学医学院附属第二医院口腔内科 [3]浙江大学医学院附属口腔医院口腔颌面外科

出　　处：《中华口腔医学杂志》2014年第1期21-26,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金

摘　　要：目的明确牙龈成纤维细胞表达蛋白酶激活受体（protease-activatedreceptors,PAR）的类型及PAR在牙周炎中的作用.方法培养原代人牙龈成纤维细胞,反转录PCR法检测PAR在牙龈成纤维细胞中的表达.以重组蛋白酶R（recombinantgingipainR,rRgp）作用于牙龈成纤维细胞,实时定量反转录PCR法检测PAR表达水平在两种rRgp（rRgpAcat和rRgpB）作用前后（rRgpAcat组和rRgpB组）的变化,并用酶联免疫吸附测定法检测细胞分泌白介素6水平的变化,以相同体积的磷酸盐缓冲液组作为对照组.应用SPSS10.0统计软件对结果数据进行分析,采用两独立样本t检验比较反转录PCR获得的PAR表达水平的差异及酶联免疫吸附测定获得的IL-6表达水平的差异,以双侧P＜0.05为差异有统计学意义.结果牙龈成纤维细胞表达PAR-1和PAR-3,二者的表达水平在rRgpAcat和rRgpB作用前后发生明显变化,PAR-1相对表达量从1.04±0.31分别下降为0.67±0.11和0.31±0.11,PAR-3相对表达量从1.01±0.44分别下降为0.79±0.13和0.44±0.12;白介素6表达水平在蛋白酶作用8h时达峰值,对照组为（18.77±4.09）μg/L,rRgpAcat和rRgpB组分别为（179.36±15.81）和（320.56±26.19）μg/L,与对照组相比差异均有统计学意义（P＜0.05）.结论牙龈成纤维细胞表达PAR-1和PAR-3,并参与牙周炎症反应,可能与牙周炎的早期急性炎症反应有关.Objective To investigate the expression types of protease-activated receptors （PAR） in human gingival fibroblasts（HGF） and the functions of PAR in periodontitis.Methods Primary HGF were cultured.Reverse transcription PCR（RT-PCR） was used to detect the expression of PAR in HGF.Recombinant gingipain R （rRgp） was applied to HGF.The change of PAR expression on the cell surface was analyzed by real-time quantitative RT-PCR,and enzyme-linked immunosorbent assay （ELISA） was used to detect the change of the interleukin（IL）-6 production from HGF.The results of RT-PCR and ELISA were statistically analyzed using the two independent samples t-test of SPSS10.0 software.Results HGF expressed PAR-1 and PAR-3.The expression of PAR-1 and PAR-3 changed after two rRgp treatment with HGF cells.The relative expression of PAR-1 was decreased from 1.04 ± 0.31 to 0.67 ± 0.11 and 0.31 ±0.11.The relative expression of PAR-3 was decreased from 1.01 ±0.44 to 0.79 ±0.13 and 0.44 ± 0.12 （P 〈 0.05）.The level of IL-6 was increased after rRgp treatment for 8 h.The control group was （18.77 ±4.09） μg/L,the rRgp treatment groups were （179.36 ± 15.81） and （320.56 ± 26.19） μg/L respectively.Conclusions HGF expressed PAR-1 and PAR-3 and were involved in periodontal inflammation.

关 键 词：成纤维细胞 受体 蛋白酶激活 白细胞介素6 牙周炎 INTERLEUKIN-6 

分 类 号：R781.4[医药卫生—口腔医学]