CAJ | 학술논문

目的研究类风湿关节炎(RA)炎性关节中高水平脂联素(AD)表达的意义.方法酶联免疫吸附试验(ELISA)测定关节液中AD及白细胞介素(IL)-1β、IL-6、IL-8、肿瘤坏死因子(TNF)-α、单核细胞趋化因子(MCP)-1和基质金属蛋白酶(MMP)-9等促炎因子/趋化因子的含量,并分析AD与其相关性.双重免疫标记分析RA滑膜中AD在滑膜成纤维细胞(SFLs)、T淋巴细胞、单核细胞、中性粒细胞和肥大细胞的表达.ELISA测定重组AD因子刺激SFLs后培养上清液中细胞因子含量变化.实时荧光定量反转录-聚合酶链反应(real-time PCR)分析AD和IL-6共刺激对人滑膜细胞系MH7A中核因子-κB受体活化因子配体(RANKL)mRNA表达影响.3组间采用多变量方差分析,2组比较采用Student's t检验.Mann-Whitney U检验,相关性分析采用Spearman检验.结果 RA关节液中AD与IL-6含量呈正相关(r=0.27,P=0.03).AD与成纤维细胞双重免疫标记呈阳性反应.以重组AD因子30g/ml刺激SFLs 24 h后,培养上清液中IL-6和MCP-1分泌量与刺激前相比分别增加了11.4倍和8倍(P<0.05).AD和IL-6共刺激后,MH7A中RANKL mRNA表达显著增加(82±26).结论炎性关节中高水平AD可促进IL-6、MCP-1和RANKL表达,其可能在RA慢性炎症和骨侵蚀过程中发挥重要作用.
목적 연구류풍습관절염(RA)염성관절중고수평지련소(AD)표체적의의.방법 매련면역흡부시험(ELISA)측정관절액중AD급백세포개소(IL)-1β、IL-6、IL-8、종류배사인자(TNF)-α、단핵세포추화인자(MCP)-1화기질금속단백매(MMP)-9등촉염인자/추화인자적함량,병분석AD여기상관성.쌍중면역표기분석RA활막중AD재활막성섬유세포(SFLs)、T림파세포、단핵세포、중성립세포화비대세포적표체.ELISA측정중조AD인자자격SFLs후배양상청액중세포인자함량변화.실시형광정량반전록-취합매련반응(real-time PCR)분석AD화IL-6공자격대인활막세포계MH7A중핵인자-κB수체활화인자배체(RANKL)mRNA표체영향.3조간채용다변량방차분석,2조비교채용Student's t검험.Mann-Whitney U검험,상관성분석채용Spearman검험.결과 RA관절액중AD여IL-6함량정정상관(r=0.27,P=0.03).AD여성섬유세포쌍중면역표기정양성반응.이중조AD인자30g/ml자격SFLs 24 h후,배양상청액중IL-6화MCP-1분비량여자격전상비분별증가료11.4배화8배(P<0.05).AD화IL-6공자격후,MH7A중RANKL mRNA표체현저증가(82±26).결론 염성관절중고수평AD가촉진IL-6、MCP-1화RANKL표체,기가능재RA만성염증화골침식과정중발휘중요작용.
Objective To explore the potential role of high levels of adiponectin (AD) in the inflammatory joint of rheumatoid arthritis (RA). Methods ELISA was used to measure the levels of AD, IL-Iβ,IL-6, IL-8, TNF-α, MCP-1 and MMP-9 in the synovial fluids of RA and osteroarthritis (OA), the levels of these cytokines were tested after the synovial fibroblasts (SFLs) were stimulated with AD. Doublelabeling immunohistochemistry was used to analyze the expression of AD in RA synovium. Cytokines were measured by ELISA after SFLs were stimulated with AD. The expression of RANKL was detected by real-time PCR after MH7A were treated with AD and IL-6 ANOVA, Student's t-test, Mann-Whitney U-tese, Spearman's-test were used for statistical analysis. Results High levels of AD in RA synovial fluids were correlated with IL-6 levels. Double labeling immunohistochemistry showed that AD was localized in fibroblasts. MCP-1 and IL-6 were dramatically increased in human synovial fibroblasts following incubation with recombinant AD for 24 h. RANKL mRNA was significantly increased in MH7A after treated with AD and IL-6. Conclusion High levels of AD in the inflammatory joints of RA are likely to contribute to the high expression of IL-6, MCP-1 and RANKL, which may play an important role in the chronic inflammation, osteoclasts activation and bone erosion in RA.