拉莫三嗪诱发皮肤不良反应与TCRBV基因表达的相关性研究  

Study on the correlation between lamotrigine-induced cutaneous adverse reactions and TCRBV genes expression

作　　者：晁丽娜 许贤瑞[2] 王旭[2] 李琪 刘美丽 张庆[2] CHAO Lina;Xu Xianrui;WANG Xu;LI Qi;LIU Meili;ZHANG Qing(Department of Neurology,General Hospital of Ningxia Medical University,Ningxia Brain Disease Laboratory,Yinchuan Ningxia 750004,China)

机构地区：[1]宁夏医科大学附属石嘴山市第一人民医院神经内科,753200 [2]宁夏医科大学总医院神经内科,宁夏颅脑疾病重点实验室,750004 [3]山东省立医院西院神经电生理科,250000 [4]河北省沧州中西医结合医院神经内二科,061000

出　　处：《中国神经免疫学和神经病学杂志》2018年第2期113-119,共7页Chinese Journal of Neuroimmunology and Neurology

基　　金：宁夏颅脑疾病重点实验室开放课题资助(XY201513);中国抗癫痫协会癫痫科研基金-UCB基金资助项目(2016001)

摘　　要：目的研究拉莫三嗪(LTG)激活T细胞介导免疫应答而诱发皮肤不良反应(cADRs)的可能致病机制;通过分析在LTG诱发的cADRs(LTG-cADRs)患者与LTG耐受者外周血单个核细胞(PBMCs)中TCRBV基因表达的差异,探讨LTG-cADRs与T细胞受体β家族V基因(TCRBV基因)表达的相关性。方法收集10例cADRs已治愈达6周以上的LTG-cADRs患者(病例组),依据病例对照研究的方法按1︰1匹配纳入10例LTG耐受者(对照组),分离病例组与对照组患者PBMCs,给予LTG体外刺激并培养,采用ELISA法测定病例组与对照组患者PBMCs分泌干扰素γ(IFN-γ)、白细胞介素5(IL-5)及肿瘤坏死因子β(TNF-β)的情况,运用实时荧光定量逆转录PCR技术(RT-qPCR)的SYBR GreenⅠ荧光染料法检测TCRBV基因的表达,比较两组IFN-γ、IL-5及TNF-β水平及TCRBV基因相对表达量的差异。结果病例组细胞上清液中IFN-γ水平明显高于对照组[分别为(1.06±0.09)ng/mL、(0.86±0.16)ng/mL,P<0.05]。病例组细胞上清液中IL-5和TNF-β水平与对照组差异无统计学意义(均P>0.05)。病例组TCRBV11、TCRBV15、TCRBV17、TCRBV18、TCRBV19基因的相对表达量[中位数(四分位数间距)]分别为1.78(1.07)、1.83(2.19)、2.13(3.39)、1.79(1.92)、1.44(1.62),均高于对照组[分别为1.00(0.43)、1.08(1.13)、1.05(0.71)、0.96(0.72)、1.01(0.61),均P<0.05]。结论 LTG可在体外激活LTG-cADRs患者的PBMCs分泌IFN-γ,提示T细胞介导的免疫反应可能参与了LTG-cADRs的致病机制。LTG-cADRs可能与TCRBV11、TCRBV15、TCRBV17、TCRBV18、TCRBV19基因的表达相关。Objective To investigate the possible pathogenesis of lamotrigine(LTG)-induced T cell-mediated immune response which caused cutaneous adverse reactions(cADRs).The differences in TCRBV genes expression were analyzed between LTG-cARDs patients and LTG tolerant patients in their peripheral blood mononuclear cells(PBMCs),to investigate the correlation between LTG-cADRs and TCRBV genes expression.Methods We collected 10 cases of cARDs whose CARDs had been cured for more than 6 weeks as a case group,and 10 LTG tolerant patients were included as the control group by 1∶1 match(according to the case-control study).We separating the PBMCs of these two groups,and gave LTG in vitro stimulation and culture.The levels of IFN-γ,IL-5 and TNF-βwhich secreted by PBMCs in patients of the case group and the control group were measured by ELISA.The expression of TCRBV genes were detected by SYBR GreenⅠfluorescent dye method using quantitative real-time reverse transcription(RT-qPCR).The differences in IFN-γ,IL-5 and TNF-βconcentrations and the relative expression of TCRBV genes were compared between the two groups.Results The concentration of IFN-γ[(1.06±0.09)ng/mL]in the supernatant of the case group was significantly higher than that in the control group[(0.86±0.16)ng/mL],the difference was statistically significant(P<0.05).The differences in IL-5 and TNF-βconcentrations was not statistically significant between the two groups(P>0.05).The median 2-ΔΔCt numerical value of TCRBV11,TCRBV15,TCRBV17,TCRBV18 and TCRBV19(the interquartile range)in the case group and the control group are 1.78(1.07)and 1.00(0.43),1.83(2.19)and 1.08(1.13),2.13(3.39)and 1.05(0.71),1.79(1.92)and 0.96(0.72),1.44(1.62)and 1.01(0.61).The relative expressions of TCRBV11,TCRBV15,TCRBV17,TCRBV18 and TCRBV19 in the case group were higher than those in the control group(P<0.05).Conclusions LTG can activate PBMCs in LTG-cARDs patients in vitro,which can secret IFN-γ,suggesting that immune responses mediated by T cell may be involved in the pathogenesis

关 键 词：拉莫三嗪 皮肤不良反应 TCRBV基因 芳香族抗癫痫药物 

分 类 号：R742.1[医药卫生—神经病学与精神病学]