甲氨蝶呤联合4-氢过氧环磷酰胺对干燥综合征患者单个核细胞的作用  

Influence by Methotrexate and 4-hydroperoxycyclophosphamide upon Sjgren Syndrome Patients' Peripheral Blood Mononuclear Cell

作　　者：霍月红[1,2] 许珂[1] 赵文鹏[1] 丁志明[1,3] 孟力平[1,3] 张莉芸[1] 李小峰[1] 

机构地区：[1]山西医科大学第二医院风湿免疫科,太原030001 [2]山西同煤集团总医院风湿科,山西大同037000 [3]山西省大同市第五医院风湿科,山西大同037000

出　　处：《中华临床免疫和变态反应杂志》2011年第3期173-178,共6页Chinese Journal of Allergy & Clinical Immunology

摘　　要：目的研究甲氨蝶呤(MTX)联合4-氢过氧环磷酰胺(4HC)对体外培养的干燥综合征(SS)患者外周血单个核细胞(PBMC)的周期、凋亡及分泌白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的影响。方法 MTX、4HC、MTX联合4HC的3组药物在4个浓度水平0、1、10、100μg/ml,与体外培养的SS患者PBMC作用48h。用流式细胞术(FCM)检测细胞周期、凋亡,用酶联免疫吸附试验(ELISA法)检测IL-1β、TNF-α分泌水平。对MTX、4HC在0、1、10μg/ml浓度水平进行析因设计。结果细胞周期:与对照组相比,MTX10组、MTX100组PBMC的G0/G1期比例增加,S期及G2/M期比例减少;不同药物浓度4HC组对细胞各周期影响多数无统计学意义;联合组中MTX1+4HC1组、MTX10+4HC10组及MTX100+4HC100组G0/G1期细胞比例减少,S期及G2/M期比例增加。细胞凋亡:MTX10组、MTX100组、4HC10组、4HC100组和联合组PBMC凋亡率高于对照组。同一药物浓度水平时,MTX1+4HC1组凋亡率高于MTX1组,MTX10+4HC10组凋亡率高于MTX10组及4HC10组,MTX100+4HC100组凋亡率高于MTX100组及4HC100组。细胞因子:不同药物浓度的MTX组、4HC组和联合组IL-1β分泌水平均低于对照组;除MTX10外,各药物组TNF-α分泌水平均低于对照组。比较同一药物浓度水平时,不同组的IL-1β、TNF-α分泌水平,混合组均低于单用药组。交互作用:MTX联合4HC在0、1、10μg/ml3个浓度对G0/G1期比例、凋亡率、IL-1β及TNF-α分泌水平4个方面的F值分别为:4.68、45.19、4.44及3.392,P值均<0.05,提示MTX和4HC存在交互作用。结论 MTX和4HC联合作用具有协同效应。MTX和4HC联合作用于体外培养的SS患者的PBMC,可有效抑制其G0/G1期的细胞凋亡,降低促炎因子IL-1β、TNF-α的分泌水平。Objective To investigate the effects of 4-hydroperoxycyclophosphamide（4HC） and methotrexate（MTX） on cell cycle,apoptosis and the secretion level of interleukin-1β（IL-1β） and tumor necrosis factor-α（TNF-α） of the peripheral blood mononuclear cell（PBMC） in the patients with Sjogren syndrome（SS）.Methods SS patients＇ PBMC was cultivated with the effect of MTX and 4HC of different levels 0,1,10,100 μg/ml alone or combined in vitro for 48 hours.Both the cell cycle and the apoptosis were analyzed by flow cytometry（FCM） and the levels of IL-1β and TNF-α were detected by ELISA.Factorial design was adopted at three concentration levels of MTX and 4HC.Results Compared with the cell cycle of the control group,the percentage of G0/G1 phase in MTX10 group and MTX100 group increased while the percentage of the S phase and G2/M phase decreased;The cell cycle distribution was not affected significantly by different concentration of 4HC.The percentage of G0/G1 phase decreased and the percentage of the S phase,G2/M phase increased in MTX1＋4HC10 group,MTX10＋4HC10 group and MTX100＋4HC100 group.Compared with the control group,the apoptosis percentage of PBMC were increased in MTX10 group,MTX100 group,4HC10 group,4HC100 group and all of the combination groups.At the same drug level,the apoptosis percentage of MTX1＋4HC1 group was higher than that of MTX10 group.And the apoptosis percentage of MTX10＋4HC10 group was higher than that of MTX10 group and 4HC10 group.At the same time,MTX100＋4HC100 group＇ s apoptosis percentage was higher than that of MTX100 group and 4HC100 group.The secretion level of IL-1β was lower in all the drug groups than that of the control group.Meanwhile,the secretion levels of TNF-α was lower in all the drug groups than that of the control group,with the exception of the MTX10 group.The secretion levels of IL-1β and TNF-α in the combination group were significantly greater than that of the groups treated by MTX or 4HC alone at the same drug level.According to f

关 键 词：干燥综合征 甲氨蝶呤 4-氢过氧环磷酰胺 细胞凋亡 白细胞介素-1Β 肿瘤坏死因子-α 

分 类 号：R593.2[医药卫生—内科学]