类风湿关节炎患者核转录因子κB表达与B细胞凋亡相关性研究  

Associations of expression of nuclear factor kappa B and apoptosis with B cell in patients with rheumatoid arthritis

作　　者：杨晋[1] 胡文霞[1] 霍娟[1] 张欢欢[1] 梁文学[1] Yang Jin ,Hu Wenxia ,Huo Juan ,Zhang Huanhuan ,Liang Wenxue(Department of Clinical Laboratory,the First People＇s Hospital of Lianyuagang,Lianyungang 222002, Chin)

机构地区：[1]连云港市第一人民医院临床实验中心,222002

出　　处：《国际免疫学杂志》2018年第2期184-189,共6页International Journal of Immunology

基　　金：连云港市科技局社会发展项目（SH1406）

摘　　要：目的探讨类风湿关节炎（rheumatoid arthritis，RA）患者外周血中B细胞核转录因子κB（nuclear factor kappa B，NF-κB）通路相关蛋白表达情况及其与B细胞凋亡的相关性。方法免疫磁珠法分选48例初诊RA患者及30例健康对照外周血中的B细胞，采用Western印迹技术，检测B细胞中NF-κB相关蛋白NF-κBp50、NF-κBp65、IκBα、磷酸化NF-κBp65（P- NF-κBp65）、磷酸化IκBα（P-IκBα）及凋亡相关蛋白Caspase-3、Bcl-2蛋白表达。酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）检测RA患者血浆中白细胞介素（interleukin，IL）-17、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平。流式细胞术检测RA患者及健康对照外周血B细胞凋亡率。两组比较用t检验，相关性分析采用Pearson相关检验。结果RA患者B细胞中P-NF-κBp65和P-IκBα蛋白表达显著高于健康对照组[分别为（30.5±10.1）比（13.4±4.8）；（25.6±9.2）比（16.5±5.8）]，差异有统计学意义（t＝5.071、t＝4.643，均P〈0.05）。RA患者血清中细胞因子IL-17、IL-6及TNF-α表达均高于健康对照组[分别为（68.4±14.2）、（30.4±8.9）、（19.2±4.1）pg/mL比（20.5±6.8）、（9.7±2.8）、（5.5±1.4）pg/mL]，差异有统计学意义（t分别为4.752、4.436、3.871，P均〈0.05）。P-NF-κBp65和P-IκBα蛋白表达与RA临床参数及相关细胞因子比较均无相关性，差异无统计学意义（P〉0.05）。RA患者中P-NF-κBp65蛋白表达与凋亡相关蛋白Bcl-2表达呈正相关（r＝0.311，P〈0.05）。RA患者P-NF-κBp65表达与B细胞凋亡率呈负相关（r＝－0.472，P〈0.05）；P-IκBα表达与B细胞凋亡率无相关性（r＝－0.075，P＝0.104）。结论RA患者外周血中B细胞P-NF-κBp65、P-IκBα表达显著增高，且P-NF-κBp65与Bcl-2表达及B细胞凋亡密切相关，提示RA中NF-κB通路活化可能调控Bcl2表达，进而抑制B细胞凋亡。ObjectiveTo investigate the relations between the expression of nuclear factor kappa B of B cells in the peripheral blood（PB） and the apoptosis with B cell in patients with rheumatoid arthritis （RA）.MethodsB cells in the PB of 48 RA and 30 healthy individuals were isolated by MACS and Western blot was used to detect the expression of NF-κBp50, NF-κBp65, IκBα, phosphor-NF-κBp65 （P-NF-κBp65）, phosphor- IκBα（P-IκBα）, apoptosis related proteins Caspase-3 and Bcl-2.The levels of interleukin（IL）-17, IL-6, tumor necrosis factor-α（TNF-α）in patients with RA were measured by enzyme-linked immunosorbent assay（ELISA）. The proportions of apoptosis of B cells in the PB of patients with RA and healthy controls were determined by flow cytometry.T-test and Pearson correlation analysis were used to analyze the data.ResultsThe expression of P-NF-κBp65 and P-IκBα of B cells in the PB of patients with RA was significantly higher than that of control group[（30.5±10.1）vs.（13.4±4.8）; （25.6±9.2）vs.（16.5±5.8）], the difference was statistically significant（t=5.071, t=4.643, both P〈0.01）. The levels of IL-17, IL-6 and TNF-α in serum of patients with RA was significantly higher than that of control group[（68.4±14.2）、（30.4±8.9）、（19.2±4.1）pg/mL vs.（20.5±6.8）、（9.7±2.8）、（5.5±1.4）pg/mL], the difference was statistically significant（t=4.752、t=4.436、t=3.871, all P〈0.05）. There was no correlation between the level of P-NF-κBp65 and P-IκBα and clinical parameters and related cytokines in RA patients（P〉0.05）. The expression of P-NF-κBp65 and Bcl-2 were positively correlated in patients with RA （r=0.311, P〈0.05）. There was a negative correlation between the level of P-NF-κBp65 and the rate of apoptosis of B cells in the patients with RA（r=-0.472, P〈0.05）. The expression of P-IκBα has no correlation with the rate of apoptosis of B cells in the patients with RA （r=-0.075, P=0.104）.ConclusionThe expression of P

关 键 词：类风湿关节炎 B细胞 核转录因子ΚB BCL-2 细胞凋亡 

分 类 号：R593.22[医药卫生—内科学]