CAJ | 학술논문

目的探讨类风湿关节炎(RA)患者外周血单核细胞高迁移率族蛋白-1(HMGBl)的释放和细胞内定位及沙立度胺对其的影响.方法抽取19例RA患者和20例健康对照者外周静脉血5 ml,Ficoll密度梯度离心法分离外周血单核细胞,分别用100 ng/ml TNFα、100 ng/ml TNFα+40 μ/md沙立度胺处理细胞后,Western blot检测外周血单核细胞HMGB1的释放;细胞免疫荧光法观察外周血单核细胞中HMGB1的胞内定位.结果 (1)在无刺激因素作用下,RA患者外周血单核细胞培养液的上清中HMGBl蛋白水平较健康对照组显著增高(P<0.05).100 ng/ml TNα并不增加RA患者外周血单核细胞中HMGBl的释放,40μg/ml沙立度胺能明显降低RA患者外周血单核细胞HMGBl的释放(P<0.05).(2)RA患者外周血单核细胞中HMGB1主要分布于胞核.100 ng/m1TNFα刺激外周血单核细胞24 h后HMGB1向胞质移位,40 μg/m1沙立度胺能明显抑制TNFα所致的HMGBl胞质移位.结论 RA患者外周血单核细胞在TNFα刺激下HMGB1从核内移位至胞质,最后释放到细胞外,沙立度胺能抑制HMGB1的胞质移位和释放.
목적 탐토류풍습관절염(RA)환자외주혈단핵세포고천이솔족단백-1(HMGBl)적석방화세포내정위급사립도알대기적영향.방법 추취19례RA환자화20례건강대조자외주정맥혈5 ml,Ficoll밀도제도리심법분리외주혈단핵세포,분별용100 ng/ml TNFα、100 ng/ml TNFα+40 μ/md사립도알처리세포후,Western blot검측외주혈단핵세포HMGB1적석방;세포면역형광법관찰외주혈단핵세포중HMGB1적포내정위.결과 (1)재무자격인소작용하,RA환자외주혈단핵세포배양액적상청중HMGBl단백수평교건강대조조현저증고(P<0.05).100 ng/ml TNα병불증가RA환자외주혈단핵세포중HMGBl적석방,40μg/ml사립도알능명현강저RA환자외주혈단핵세포HMGBl적석방(P<0.05).(2)RA환자외주혈단핵세포중HMGB1주요분포우포핵.100 ng/m1TNFα자격외주혈단핵세포24 h후HMGB1향포질이위,40 μg/m1사립도알능명현억제TNFα소치적HMGBl포질이위.결론 RA환자외주혈단핵세포재TNFα자격하HMGB1종핵내이위지포질,최후석방도세포외,사립도알능억제HMGB1적포질이위화석방.
Objective To investigate the release and intracellular localization of high mobility group box chromosomal protein 1(HMGBl)in the peripheral blood monocytes of rheumatoid arthritis(RA) patients and the inhibitive effect of thaiidomide.Methods 19 RA patients and 20 healthy controls were included in the study.Monocytes were separated from peripheral blood with Ficoll density gradient centrifugation.Monocytes were treated with 100 ng/ml tumor necrosis factor α(TNFa)or 100 ng/ml TNFα plus 40 μg/ml thalidomide and grown in an incubator at 37℃ with 5%CO,for 24 hours.The cuIture supernatants of the monocytes were collected.HMGB1 level in the culture medium was detected with Western blot.In addition,the intraceUular localization of HMGB1 in the fflonocytes was investigated with immunocytochemical analysis. Results Without stimulation. the release of HMGBl protein was significantly increased in the culture supernatants of peripheral blood monocytes from RA patients as compared with that from healthy controls(P<0.05).TNFα(100 ng/ml)did not further increase the release of HMGBl in the monocytes from the patients with RA.Thalidomide(40 μg/ml)could inhibit the release of HMGB1 in the monocytes from RA patients stimulated with TNFα(P<0.05).In the monocytes from RA patients,HMGBl was mainly localized in the nucleus.Treatment with TNFOL(100 ng/ml)for 24 hour resulted in a cytoplasmic translocation of HMGB1,which was inhibited significantly by thalidomide. Conclusion TNFα induces the release and cytoplasmic translocation of HMGBI in the peTipheral blood monocytes of RA patients and thalidomide inhibits the release and translocation of HMGB1.