牙龈卟啉单胞菌肽酰精氨酸脱亚氨酶与类风湿性关节炎的相关机制研究

项目名称： 牙龈卟啉单胞菌肽酰精氨酸脱亚氨酶与类风湿性关节炎的相关机制研究

项目编号： No.81500869

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 刘静波

作者单位： 中国医科大学

项目金额： 17万元

中文摘要： 类风湿性关节炎(RA)成纤维样滑膜细胞(FLS)的增殖和迁徙在RA发生发展过程中起重要作用。牙龈卟啉单胞菌(P.g)是牙周病炎重要致病菌，在RA FLS及关节滑液中检出率高。PG1424基因编码肽酰精氨酸脱亚氨酶(PPAD),PPAD的功能是学者研究RA与牙周炎相关关系的焦点,但机制不清存在争议。前期实验发现P.g内化于RA FLS后，PG1424基因上调，细胞RhoA表达上调，促进RA FLS增殖，结果提示PPAD及RhoA参与P.g促进RA FLS增殖过程。我们推测“PPAD通过RhoA/ROCK信号转导通路调控RA FLS增殖、迁徙等细胞生物学行为”是P.g和RA关联的机制之一。本研究拟构建PG1424突变菌株及回复突变菌株，制备细菌与RA FLS共培养模型和细菌感染RA动物模型，应用siRNA、PCR芯片等技术验证假说，为牙周炎和RA的预防和治疗提供新思路。

中文关键词： 牙龈卟啉单胞菌；肽酰精氨酸脱亚氨酶；；慢性牙周炎；类风湿性关节炎

英文摘要： The proliferation and migration of rheumatoid arthritis fibroblast-like synoviocytes (RA FLS) play an important role in development process in RA. Porphyromonas gingivalis (P. gingivalis), a Gramnegative anaerobic black-pigmented rod, is one of the major pathogens associated with periodontitis. P. gingivalis is the only known oral bacterium expressiong the Peptidylarginine deaminase (PAD, coded by PG1424), which leads to the citrullination of rheumatoid arthritis (RA) autoantigen such as fibrin in synovium joint. Moreover, P. gingivalis has also been identified in fibroblast-like synoviocytes (RA FLS) and synovial fluid from patients with RA. Recent findings suggest a causative link between periodontitis and RA via P. gingivalis-dependent induction, but the role of PPAD is controversial. Our previous study found P. gingivalis invaded RA FLS, then increased PG1424 of P. gingivalis and RhoA of RA FLS mRNA expression, promoted the RA FLS cell proliferation, but the regulation mechanism was still unknown. We speculate that PPAD regulate cell biology behavior of RA FLS via RhoA/ROCK signaling pathways, which is a new mechanism of P.g associated with RA. In this study, we will 1) create the PG1424-defective mutant and revertant by allelic exchange, and express and purify P. gingivalis PAD (PPAD); 2) constructe the co-culture model of RA FLS with PG1424-defective mutant or PPAD, then study that PPAD modulate RA FLS’s biological behavior including proliferation, apoptosis, changes of cell cycle, migration, and cellular morpholog via the RhoA/ROCK pathway by inhibiting and activating RhoA/ROCK, and using PCR array, Human Cytokine Antibody Array 1 and bioinformatics technology; 3) constructe RA rat model, analyze that PPAD’s influence on RA rat by using PG1424-defective mutant or revertant infection RA rat model. This study will provide theoretical basis for elaborating the associated mechanism of periodontitis and RA, and raise the new ideas for preventing and treating of periodontitis and RA.

英文关键词： Porphyromonas gingivalis;Peptidylarginine deaminase;Chronic Periodontitis;Rheumatoid Arthritis