中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2015年
8期
552-556
,共5页
刘俊文%刘红春%苏利沙%邓少丽%张根豪%张俊华%魏高辉
劉俊文%劉紅春%囌利沙%鄧少麗%張根豪%張俊華%魏高輝
류준문%류홍춘%소리사%산소려%장근호%장준화%위고휘
关节炎,类风湿%白细胞介素类%阻遏蛋白质类%白细胞,单核%CD40配体
關節炎,類風濕%白細胞介素類%阻遏蛋白質類%白細胞,單覈%CD40配體
관절염,류풍습%백세포개소류%조알단백질류%백세포,단핵%CD40배체
Arthritis,rheumatoid%Interleukins%Repressor proteins%Leukocytes,mononuclear%CD40 ligand
目的:研究IL-21、Blimp1 mRNA在类风湿关节炎( RA)患者体内的表达及IL-21刺激后对体外培养RA患者外周血单个核细胞( PBMCs ) Blimp1表达的影响,探讨IL-21、Blimp1参与RA发病的具体作用机制。方法病例对照研究。收集2012年10月至2013年3月郑州大学第一附属医院住院确诊的RA患者50例和同期健康体检者50名为对照组的外周静脉血,分离血浆及PBMC, ELISA检测血浆IL-21水平,同时检测患者临床指标DAS28和抗环瓜氨酸肽抗体(抗CCP )抗体将其与患者IL-21水平进行相关性分析。实时荧光定量PCR ( qPCR )检测患者PBMC Blimp1 mRNA表达量;分离RA患者PBMCs进行体外培养,经IL-21和CD40L刺激细胞72 h后,检测患者PBMC Blimp1 mRNA表达量,流式细胞术检测各组细胞CD20阳性B细胞和CD138阳性细胞所占比例。采用均值t检验、Wilcoxon秩和检验和方差分析等方法进行统计学分析。结果 RA 患者血清 IL-21水平(130.51±11.35)ng/L明显高于健康对照组(25.46±6.05)ng/L,t=5.39,P=0.007,且IL-21水平与RA患者DAS289(r=0.658)和抗CCP抗体(r=0.674)具有相关性(P分别为0.019、0.016)。 RA患者PBMCs Blimp1 mRNA表达水平(1.321±0.110)高于健康对照组(1.000±0.000),Z=-2.48,P<0.05。 IL-21及CD40L体外刺激后,IL-21组和CD40L+IL-21组Blimp1 mRNA表达量分别为(1.084±0.029)、(1.157±0.028),高于对照组(1.000±0.000),P 分别为0.002、0.001,CD40L +IL-21组Blimp1mRNA表达水平高于IL-21组(t=4.862,P=0.02);CD40L组、IL-21组及IL-21+CD40L组较阴性对照组CD20阳性B细胞比例增加[2.42±0.35、2.63±0.33、6.35(4.85,6.57),F=278.363, P<0.001],CD138阳性细胞所占比例也增加(0.474±0.110、0.668±0.120、0.955±0.170,F =49.01,P<0.001),差异均具有统计学意义。结论 IL-21可促进RA患者外周单个核细胞Blimp1 mRNA的表达;IL-21和CD40L协同作用可能通过调节Blimp1 mRNA的表达促进B细胞的分化成熟进而参与RA的发病。(中华检验医学杂志,2015,38:552-556)
目的:研究IL-21、Blimp1 mRNA在類風濕關節炎( RA)患者體內的錶達及IL-21刺激後對體外培養RA患者外週血單箇覈細胞( PBMCs ) Blimp1錶達的影響,探討IL-21、Blimp1參與RA髮病的具體作用機製。方法病例對照研究。收集2012年10月至2013年3月鄭州大學第一附屬醫院住院確診的RA患者50例和同期健康體檢者50名為對照組的外週靜脈血,分離血漿及PBMC, ELISA檢測血漿IL-21水平,同時檢測患者臨床指標DAS28和抗環瓜氨痠肽抗體(抗CCP )抗體將其與患者IL-21水平進行相關性分析。實時熒光定量PCR ( qPCR )檢測患者PBMC Blimp1 mRNA錶達量;分離RA患者PBMCs進行體外培養,經IL-21和CD40L刺激細胞72 h後,檢測患者PBMC Blimp1 mRNA錶達量,流式細胞術檢測各組細胞CD20暘性B細胞和CD138暘性細胞所佔比例。採用均值t檢驗、Wilcoxon秩和檢驗和方差分析等方法進行統計學分析。結果 RA 患者血清 IL-21水平(130.51±11.35)ng/L明顯高于健康對照組(25.46±6.05)ng/L,t=5.39,P=0.007,且IL-21水平與RA患者DAS289(r=0.658)和抗CCP抗體(r=0.674)具有相關性(P分彆為0.019、0.016)。 RA患者PBMCs Blimp1 mRNA錶達水平(1.321±0.110)高于健康對照組(1.000±0.000),Z=-2.48,P<0.05。 IL-21及CD40L體外刺激後,IL-21組和CD40L+IL-21組Blimp1 mRNA錶達量分彆為(1.084±0.029)、(1.157±0.028),高于對照組(1.000±0.000),P 分彆為0.002、0.001,CD40L +IL-21組Blimp1mRNA錶達水平高于IL-21組(t=4.862,P=0.02);CD40L組、IL-21組及IL-21+CD40L組較陰性對照組CD20暘性B細胞比例增加[2.42±0.35、2.63±0.33、6.35(4.85,6.57),F=278.363, P<0.001],CD138暘性細胞所佔比例也增加(0.474±0.110、0.668±0.120、0.955±0.170,F =49.01,P<0.001),差異均具有統計學意義。結論 IL-21可促進RA患者外週單箇覈細胞Blimp1 mRNA的錶達;IL-21和CD40L協同作用可能通過調節Blimp1 mRNA的錶達促進B細胞的分化成熟進而參與RA的髮病。(中華檢驗醫學雜誌,2015,38:552-556)
목적:연구IL-21、Blimp1 mRNA재류풍습관절염( RA)환자체내적표체급IL-21자격후대체외배양RA환자외주혈단개핵세포( PBMCs ) Blimp1표체적영향,탐토IL-21、Blimp1삼여RA발병적구체작용궤제。방법병례대조연구。수집2012년10월지2013년3월정주대학제일부속의원주원학진적RA환자50례화동기건강체검자50명위대조조적외주정맥혈,분리혈장급PBMC, ELISA검측혈장IL-21수평,동시검측환자림상지표DAS28화항배과안산태항체(항CCP )항체장기여환자IL-21수평진행상관성분석。실시형광정량PCR ( qPCR )검측환자PBMC Blimp1 mRNA표체량;분리RA환자PBMCs진행체외배양,경IL-21화CD40L자격세포72 h후,검측환자PBMC Blimp1 mRNA표체량,류식세포술검측각조세포CD20양성B세포화CD138양성세포소점비례。채용균치t검험、Wilcoxon질화검험화방차분석등방법진행통계학분석。결과 RA 환자혈청 IL-21수평(130.51±11.35)ng/L명현고우건강대조조(25.46±6.05)ng/L,t=5.39,P=0.007,차IL-21수평여RA환자DAS289(r=0.658)화항CCP항체(r=0.674)구유상관성(P분별위0.019、0.016)。 RA환자PBMCs Blimp1 mRNA표체수평(1.321±0.110)고우건강대조조(1.000±0.000),Z=-2.48,P<0.05。 IL-21급CD40L체외자격후,IL-21조화CD40L+IL-21조Blimp1 mRNA표체량분별위(1.084±0.029)、(1.157±0.028),고우대조조(1.000±0.000),P 분별위0.002、0.001,CD40L +IL-21조Blimp1mRNA표체수평고우IL-21조(t=4.862,P=0.02);CD40L조、IL-21조급IL-21+CD40L조교음성대조조CD20양성B세포비례증가[2.42±0.35、2.63±0.33、6.35(4.85,6.57),F=278.363, P<0.001],CD138양성세포소점비례야증가(0.474±0.110、0.668±0.120、0.955±0.170,F =49.01,P<0.001),차이균구유통계학의의。결론 IL-21가촉진RA환자외주단개핵세포Blimp1 mRNA적표체;IL-21화CD40L협동작용가능통과조절Blimp1 mRNA적표체촉진B세포적분화성숙진이삼여RA적발병。(중화검험의학잡지,2015,38:552-556)
Objective To Investigate the expression of IL-21 and Blimp1 mRNA in Rheumatoid arthritis ( RA) patients and the influence on the expression of Blimp 1 in peripheral blood mononuclear cells (PBMCs) of RA patients after IL-21 stimulated; To further explore the mechanism of IL-21 and blimp1 in the pathogenesis of RA.Methods Case control study.The samples of peripheral venous blood from 50 RA patients of department of rheumatology of The First Affiliated Hospital of Zhengzhou University and 50 healthy people were collected respectively , then the plasma and PBMCs was separated.IL-21 in plasma was measured by ELISA; the correlation between patients clinical index DAS 28, anti-CCP antibody and IL-21 was analyzed.Blimp1 mRNA of patients′PBMCs was detected by qPCR; PMBCs were isolated from RA patient and then cultured in vitro.Blimp1 mRNA level was measured by qPCR and the ratio of CD 20 positive B cell and the ratio of CD138 positive cells in all groups were detected by flow cytometry after 72 h stimulated by IL-21 and CD40L.Results IL-21 content in RA patient blood plasma (130.51 ±11.35)ng/L was significantly higher than that in healthy control (25.46 ±6.05)ng/L, t=5.39,P<0.05.Besides, IL-21 level also had a close relativity with patients DAS28(r=0.658) and anti-CCP antibody (r=0.674, P=0.039 and 0.035).In addition, the expression level of Blimp1 mRNA in RA patient PMBCs (1.321 ± 0.11)was higher than that in healthy control group (1.000 ±0.000), Z=-2.48, P<0.05.While after IL-21 and/or CD40L stimulation, Blimp1 mRNA of IL-21 group and CD40L+IL-21 group(1.084 ±0.029, 1.157 ±0.028)were higher than those of control (1.000 ±0.000)(P=0.002,P=0.001), moreover the expressive level of Blimp1mRNA of CD40L+IL-21 group was higher than that of control group (t=4.862, P=0.02).Compared to negative control group , the ratio of CD20 positive B cells [2.42 ±0.35, 2.63 ± 0.33, 6.35(4.85,6.57),F=278.363,P<0.001] and the ratio of CD138 positive cells(0.474 ±0.110, 0.668 ±0.120, 0.955 ±0.170,F=49.01, P<0.001) in CD40L group, IL-21 group and CD40L+IL-21 group were much higher and the differences among CD 40L+IL-21 group with CD40L group and IL-21 group were statistically significant.Conclusion IL-21 could promote the level of Blimp 1 mRNA in peripheral blood mononuclear cells in RA patient; IL-21 and CD40L could co-promote B cell maturation though regulating Blimp1 mRNA expression and eventually participate in RA pathogenesis.