来氟米特对EAE豚鼠外周血CD28/CTLA-4共刺激分子表达的影响  

Effect of leflunomide on peripheral blood CD28/CTLA-4 costimulatory molecules expression in guinea pig with EAE

作　　者：吕志宇[1] 袁正洲[1] 荣本兵[1] 李作孝[1] 

机构地区：[1]泸州医学院附属医院神经内科,四川泸州646000

出　　处：《中国生化药物杂志》2015年第6期39-42,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：泸州医学院自然科学科研项目(09012)

摘　　要：目的通过观察来氟米特对实验性变态反应性脑脊髓炎(experimental allergic encephalomyelitis,EAE)豚鼠外周血CD28/CTLA-4共刺激分子表达的影响,探讨来氟米特对EAE的防治作用及免疫学机制。方法将50只成年雌性豚鼠随机分为5组,每组各10只,分别为正常对照组、EAE对照组、低剂量、治疗组、中剂量治疗组和高剂量治疗组。低、中、高剂量治疗组于造模前7 d始按体质量分别予来氟米特10、20、40 mg/(kg·d)灌胃,均1次/天,至实验终止。流式细胞术检测各组外周血CD4+T细胞膜上CD28、CTLA-4共刺激分子表达情况并记录豚鼠发病情况。结果高、中剂量治疗组较EAE对照组潜伏期延长、进展期缩短、神经功能障碍评分降低,差异有统计学意义(P<0.05),以高剂量治疗组最明显(P<0.05)。EAE对照组较正常对照组发病高峰期外周血CD4+T细胞膜上CD28分子表达升高(P<0.05),而CTLA-4表达下降(P<0.05);高、中剂量治疗组较EAE对照组CD28分子表达下调(P<0.05),而CTLA-4表达升高(P<0.05);各治疗组间CD28分子的表达以高剂量治疗组下调最显著(P<0.05),CTLA-4分子的表达以高剂量治疗组上调显著(P<0.05)。相关分析显示EAE对照组、各治疗组外周血CD28/CTLA-4比值与神经功能障碍评分均呈正比(r=0.85,P=0.002;r=0.77,P=0.000)。结论来氟米特可减轻EAE豚鼠神经功能缺损症状,且剂量越高效果越好。其作用机制可能是通过下调外周血CD4+T细胞上CD28分子,上调CTLA-4分子的表达,使EAE豚鼠免疫失衡重新达到平衡,从而减轻炎性反应,调节免疫应答,减轻临床症状。Objective To investigate immunological mechanism underlying leflunomide （ LEF ） against experimental allergic encephalomyelitis （ EAE） by studying the effects of LEF on peripheral blood CD28/CTLA-4 costimulatory molecules expression in guinea pig with EAE.Methods 50 adult female guinea pigs were randomly divided into normal control group,low dose group,EAE control group,middle dose group,and high dose group, 10 guinea pigs in each group respectively.Low, medium and high dose group were treated with LEF 10 mg/kg· D, 20 mg/kg· D and 40 mg/kg· D orally, 1 times/day, to the termination of the experiment.The expression of CD28, CTLA-4 were detected by flow cytometry, incidence was recorded at the same time.Results Compared with EAE control group,high dose and middle dose group incubation period were prolonged, progress period were shorten and neurological dysfunction score decreased（P〈0.05）.Compared with normal control group, the expression of CD28 of EAE control group increased and the decreased expression of CTLA-4 （P〈0.05）.High, middle dose treatment group compared with EAE control group CD28 down regulated expression （P〈0.05）, but the expression of CTLA-4 increased（P〈0.05）.Among the treatment groups, of CD28 molecules with high dose group decreased significantly（P〈0.05）, the expression of CTLA-4 molecules with high dose treatment groups upregulated significantly （P〈0.05）.Correlation analysis showed that the EAE control group, the treatment group the CD28/CTLA-4 ratio in peripheral blood and nerve dysfunction score is proportional （ r=0.85, P=0.002; r=0.77, P=0.000）.Conclusion LEF can reduce EAE guinea pig neurological symptoms, the better and the higher dose effect.Its mechanism may be through down-regulation of CD28 molecules in peripheral blood, upregulation of the expression of CTLA-4 molecule.so as to reduce the inflammatory response, relieve the clinical symptoms.

关 键 词：来氟米特 实验性变态反应性脑脊髓炎 多发性硬化 共刺激分子 CD28 CTLA-4 

分 类 号：R556[医药卫生—血液循环系统疾病]