CAJ | 학술논문

目的探讨自然杀伤(NK)p44+ NK细胞在类风湿关节炎(RA)滑膜增生及炎症中的作用及机制.方法采用流式细胞术检测50例RA患者及50名健康对照者外周血、30例RA患者滑液中NKp44+ NK细胞比例.流式细胞术分选5例RA患者滑液NKp44+ NK细胞(108/L),ELISA检测NKp44+ NK细胞培养上清白细胞介素(IL)-22浓度.NKp44+ NK细胞培养上清作用于RA成纤维样滑膜细胞(FLS)24、48及72 h,MTT法检测FLS增殖.rhIL-22作用RA FLS72h后检测单核细胞趋化蛋白(MCP)-1分泌.结果 (1)RA患者外周血NKp44+ NK细胞比例为1.270％,高于健康对照外周血NKp44+ NK细胞比例0(P ＜0.01);RA患者滑液NKp44+ NK细胞比例为15.190％,高于自身外周血NKp44+ NK细胞比例2.425％ (P ＜0.01).(2)RA滑液NKp44+ NK细胞体外培养2周,其上清IL-22浓度(1 603±332) ng/L.(3)NKp44+NK细胞上清作用于RA FLS 24、48、72 h均能刺激RA FLS增殖(P＜0.叭).IL-22抗体能明显抑制NKp44+ NK细胞上清诱导的RA FLS增殖(P＜0.01).(4)1及10 μg/LrhIL-22作用于RA FLS 72 h可促进FLS MCP-1分泌(P＜0.01).结论 NKp44+ NK细胞能分泌IL-22促进RA FLS增殖及MCP-1分泌,可能在RA滑膜增生及炎症中具有重要作用.
목적 탐토자연살상(NK)p44+ NK세포재류풍습관절염(RA)활막증생급염증중적작용급궤제.방법 채용류식세포술검측50례RA환자급50명건강대조자외주혈、30례RA환자활액중NKp44+ NK세포비례.류식세포술분선5례RA환자활액NKp44+ NK세포(108/L),ELISA검측NKp44+ NK세포배양상청백세포개소(IL)-22농도.NKp44+ NK세포배양상청작용우RA성섬유양활막세포(FLS)24、48급72 h,MTT법검측FLS증식.rhIL-22작용RA FLS72h후검측단핵세포추화단백(MCP)-1분비.결과 (1)RA환자외주혈NKp44+ NK세포비례위1.270％,고우건강대조외주혈NKp44+ NK세포비례0(P ＜0.01);RA환자활액NKp44+ NK세포비례위15.190％,고우자신외주혈NKp44+ NK세포비례2.425％ (P ＜0.01).(2)RA활액NKp44+ NK세포체외배양2주,기상청IL-22농도(1 603±332) ng/L.(3)NKp44+NK세포상청작용우RA FLS 24、48、72 h균능자격RA FLS증식(P＜0.팔).IL-22항체능명현억제NKp44+ NK세포상청유도적RA FLS증식(P＜0.01).(4)1급10 μg/LrhIL-22작용우RA FLS 72 h가촉진FLS MCP-1분비(P＜0.01).결론 NKp44+ NK세포능분비IL-22촉진RA FLS증식급MCP-1분비,가능재RA활막증생급염증중구유중요작용.
Objective To explore the effects of NKp44 + NK cells from rheumatoid arthritis (RA) patients on the proliferation and monocyte chemotactic protein 1 production of fibroblast-]ike synoviocytes (FLS).Methods The proportions of natural killer (NK) p44 N K cells in peripheral blood (PB) of 50 RA patients and 50 healthy individuals were detected by flow cytometry.Synovial fluid (SF) samples from 30 RA patients were also detected.NKp44 + NK cells in RA SF were sorted by flow cytometry for 5 times.The supernatant level of interleukin (IL)-22 was measured by enzyme-linked immunosorbent assay (ELISA).The proliferation of FLS after an addition of culture supernatant of NKp44 + NK cells was detected by methyl thiazolyl tetrazolium (MTT) at 24,48 and 72 h.Monocyte chemotactic protein (MCP)-1 production of RA FLS after an addition of rhIL-22 was detected by ELISA.Results The proportion of NKp44 + NK cells in PB of RA patients was significantly higher than that of normal controls while the proportion of NKp44 + NK cells in SF of RA patients was higher than that in PB of matched RA patients (1.270％ vs 0,15.190％ vs 2.425％,P ＜0.01).The supernatant level of IL-22 in NKp44 + NK cell culture was (1 603 ±332) ng/L.Rapid proliferation of RA FLS was observed at 24,48,72 h after an addition of culture supernatant(P ＜ 0.01).IL-22 antibody obviously inhibited the proliferation of RA FLS induced by NKp44 + NK cells.MCP-1 production of RA FLS was detected at 72 h after an addition of rhIL-22(P ＜0.01).Conclusion NKp44 + NK cells can promote the proliferation and MCP-1 production of RA FLS through the production of IL-22 so as to play an important role in the synovial proliferation and inflammation of RA.