周期联合甲氨蝶呤和环磷酰胺治疗诱导型干燥综合征小鼠及对涎腺B淋巴细胞刺激因子表达影响的研究  

Effect of combined methotrexate and cyclophosphamide therapy on the expression of B cell activation factor belonging to the TNF family in the salivary glands of mice with induced Sjogren＇s syndrome

作　　者：许珂[1] 赵文鹏[1] 霍月红[1] 魏华[1] 李芳[1] 张琳[1] 张奉春[2] 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]中国医学科学院北京协和医院风湿免疫科

出　　处：《中华风湿病学杂志》2009年第9期603-607,共5页Chinese Journal of Rheumatology

基　　金：山西医科大学青年基金（02200714）;山西医科大学第二医院博士启动基金（20080407）

摘　　要：目的通过研究B淋巴细胞刺激因子（BAFF）在诱导型干燥综合征（SS）小鼠涎腺的表达以及周期联合免疫抑制剂对其影响，探讨甲氨蝶呤（MTX）与环磷酰胺（CTX）联合治疗SS可能的协同作用机制。方法采用外分泌腺混合匀浆抗原诱导免疫雌性C57BL／6小鼠，建立SS模型，随机分为模型对照组、小剂量MTX治疗组（0．02m彰周）、大剂量MTX治疗组（0．06mg／周）、CTX间歇治疗组（每3周1．2mg）、CTX隔日治疗组（每2d0．6mg）、联合治疗组（MTX每周0．02mg＋CTX每3周1．2mg）。治疗18周后进行唾液分泌评估、观察靶器官损害情况、测定外分泌腺体内BAFF蛋白表达、BAFF mRNA水平。结果18周后小剂量MTX治疗组唾液分泌量增加最少，接近模型对照组水平；CTX隔日治疗组和联合治疗组增加趋势较明显。免疫组织化学法检测浸润的炎性细胞及导管上皮细胞高表达BAFF蛋白，模型对照组呈强阳性，小剂量MTX组、大剂量MTX组、CTX间歇治疗组呈中等强度阳性，联合治疗组和CTX隔日治疗组弱染；不同组间免疫组织化学灰度值差异有统计学意义（P〈0．01）。聚合酶链反应（PCR）法检测各治疗组间BAFF水平差异有统计学意义（P〈0．01）。MTX、CTX对BAFF的影响有交互作用（P〈0．05）。结论MTX和CTX可以恢复ss小鼠模型腺体功能、改善腺体病理改变、抑制BAFF表达，可能通过影响BAFF信号途径发挥治疗作用；MTX与CTX对SS小鼠模型治疗存在药物交互作用，且为协同效应。Objective To investigate the effect of combination therapy by observing the salivary glands function and related organ pathology after given methotrexate （MTX） and eyclophosphamide （CTX） intermittently in induced mice model of Sjogren＇s syndrome （SS）. To further explore the synergistic effect of combination therapy by detecting the immunological regulatory factor B cell activation factor belonging to the TNF family （BAFF） expression. Methods The ingredients of Lewis rat＇s exocrine glands homogenate were injected into female C57BL/6 mice to set up the mice model of SS. After established the SS mice model successfully, they were randomly divided into six SS model group, including low-dose MTX treatment group （0.02 mg/w）, high-dose MTX treatment group （0.06 mg/w）, CTX pause treatment group （1.2 mg/3 w）, CTX alternate day treatment group （0.6 mg/2 d）, MTX＋CTX combination treatment group （MTX 0.02 mg/3 w ＋ CTX 1.2 rag/3 w）. Treatment effects were assessed both clinically and histologically. Results Eighteen weeks after the first treatment, the improvement of the salivary secretion of the CTX alternate day treatment group and MTX＋CTX combination treatment group were higher than other groups, which showed statistically significant difference （P〈0.01）. Compared with the SS model control group, HE staining showed that the lymphocyticinfiltration of exocrine glands was decreased in the treatment group. In the CTX alternate day treatment group and MTX＋CTX combination treatment group, few amount of inflammatory cell infiltration were found, and the expression intensity of BAFF mRNA and protein were decreased markedly in salivary gland than others by RT-PCR and immunohistochemistry assay （P〈0.01）. Conclusion MTX and CTX can inhibit lymphocytic infiltration of the salivary glands, inhibit BAFF transcriptional level and production of BAFF protein, leading to an increase of fluid production. It suggests that modulation of signaling via BAFF pathways may be a mechanism o

关 键 词：干燥综合征 涎腺 甲氨蝶呤 环磷酰胺 

分 类 号：R378.2[医药卫生—病原生物学] R392.11[医药卫生—基础医学]