赵宁?侯成成?章鹏?于仲涛?杨东星?卢伟伟?沈爱娟?刘铁?田俊阁?于笑霞

【摘要】目的 研究核苷酸结合寡聚化结构域1（NOD1）和NOD2在类风湿关节炎（RA）患者外周血中的表达水平，初步探讨NOD1和NOD2在RA疾病中的作用。方法 采集40例RA患者（RA组）、40例骨关节炎（OA）患者（OA组）及40名健康志愿者（對照组）的外周血标本，分离外周血单个核细胞（PBMC），使用荧光定量PCR（qPCR）检测3组PBMC中NOD1和NOD2 mRNA水平。随机抽取10例RA患者、10例OA患者及10名健康志愿者的PBMC加入胎牛血清培养，并加入胞壁酰二肽（MDP）培养24 h，ELISA法检测3组上清液中IL-1β、IL-6、IL-8、TNF-α水平。结果 3组PBMC中NOD2 mRNA的表达水平比较差异有统计学意义（P < 0.001），其中RA组PBMC中NOD2 mRNA表达水平高于其他2组（P均< 0.001），而3组PBMC中NOD1 mRNA表达水平比较差异无统计学意义（P > 0.05）。MDP作用于3组PBMC后，RA组的IL-1β、IL-6、IL-8、TNF-α水平均高于另外2组（P均< 0.05），而OA组与对照组比较差异均无统计学意义（P均> 0.05）。结论 RA患者的NOD2 mRNA表达水平较高。NOD2激活可能诱发IL-1β、IL-6、IL-8、TNF-α等促炎因子释放。NOD2可能参与了RA的发病过程。

【关键词】类风湿关节炎;核苷酸结合寡聚化结构域;外周血单核细胞;促炎因子

Expression of NOD-like receptors in peripheral blood of patients with rheumatoid arthritis Zhao Ning， Hou Chengcheng， Zhang Peng， Yu Zhongtao， Yang Dongxing， Lu Weiwei， Shen Aijuan， Liu Tie， Tian

Junge， Yu Xiaoxia. Department of Rheumatology and Immunology， Hebei Province Cangzhou Hospital of Integ-rated Traditional and Western Medicine， Cangzhou 061000， China

Corresponding author， Hou Chengcheng， E-mail： m15055308511@ 163. com

【Abstract】Objective To investigate the expression levels of nucleotide-binding oligomerization domain 1 （NOD1） and nucleotide-binding oligomerization domain 2 （NOD2） mRNA in the peripheral blood of patients with rheumatoid arthritis （RA）， aiming to explore the roles of NOD1 and NOD2 in RA disease. Methods The expression levels of NOD1 and NOD2 mRNA in the peripheral blood mononuclear cells （PBMCs） of 40 patients with RA， 40 osteoarthritis （OA） and 40 healthy volunteers were detected by real-time PCR （qPCR）. PBMCs randomly selected from 10 RA， 10 OA patients and 10 healthy volunteers were supplemented with fetal bovine serum and stimulated by muramyl dipeptide （MDP） for 24 h. The levels of IL-1β， IL-6， IL-8 and TNF-α in the supernatant were detected by ELISA.  Results The expression levels of NOD2 mRNA in PBMCs of significantly differed among three groups （all P < 0.001）. The NOD2 mRNA level in the RA group was significantly higher than those in the other two groups （both P < 0.001）. However， there was no significant difference in the expression of NOD1 mRNA among three groups （P > 0.05）. After adding MDP to the PBMCs， the expression levels of IL-1β， IL-6， IL-8 and TNF-α in the supernatant of PBMCs of RA patients were significantly higher than those in the other two groups （all P < 0.05）， whereas there was no significant difference between the OA and the healthy volunteer groups （all P > 0.05）.  Conclusions The expression level of NOD2 mRNA in PBMCs of RA patients is higher than those of OA patients and healthy volunteers. The activation of NOD2 can induce the release of pro-inflammatory factors， such as IL-1β， IL-6， IL-8 and TNF-α， suggesting that NOD2 may be involved in the pathogenesis of RA.

【Key words】Rheumatoid arthritis;Nucleotide-binding oligomerization domain;

Peripheral blood mononuclear cell;Pro-inflammatory cytokine

類风湿关节炎（RA）是一种慢性、全身性自身免疫性疾病，以侵蚀性、对称性多关节炎为主要临床表现。虽然RA的确切发病机制尚未明确，但既往研究显示固有免疫的激活在该病的发病中起着重要作用[1]。固有免疫系统是机体抵抗病原微生物和内在损伤组织入侵的第一道防线，模式识别受体（PRR）是固有免疫系统中的重要成员，其可以识别病原相关分子模式（PAMP）和损害相关分子模式（DAMP）产生相应的对抗病原微生物及损害分子的固有免疫反应。 PRR主要包括Toll样受体（TLR）和细胞内核苷酸结合寡聚化结构域（NOD）蛋白家族中的NOD样受体（NLRs）。NOD1和NOD2是NLR中最为重要且研究较多的受体[2-3]。既往研究显示，NOD1和NOD2激活后可导致多条信号通路的激活（包括NF-κB及MAPK通路），进而释放多种促炎因子，参与自噬过程;此外其还与TLR有交叉协同作用[4]。……