来氟米特对NOD鼠颌下腺唾液分泌功能的保护作用  

Protective effects of leflunomide on salivary function of submandibular gland in NOD mice

作　　者：杨超 郑旭 杨晓晓 汪国生 李向培 厉小梅 Yang Chao;Zheng Xu;Yang Xiaoxiao;Wang Guosheng;Li Xiangpei;Li Xiaomei(Department of Rheumatology and Immunology,Anhui Provincial Hospital Affiliated to Anhui Medical University,Hefei 230001,China;Department of Rheumatology and Immunology,the First Affiliated Hospital of University of Science and Technology of China,Hefei 230001,China)

机构地区：[1]安徽医科大学附属省立医院风湿免疫科,合肥230001 [2]中国科学技术大学附属第一医院风湿免疫科,合肥230001

出　　处：《中华微生物学和免疫学杂志》2020年第3期211-217,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目(81871271);安徽省重点研究与开发计划(1804b06020354)。

摘　　要：目的:探讨来氟米特对干燥综合征模型NOD鼠唾液腺分泌功能减退的治疗作用。方法:将NOD鼠随机分配为4组:预防用药组、预防对照组、治疗用药组、治疗对照组;检测毛果芸香碱刺激后唾液流率变化;苏木精和伊红染色后比较平均浸润灶数目及面积的变化;流式细胞术检测颌下腺、脾脏组织中CD3 +T、CD4 +T、CD8 +T、CD44 +CD62L -CD4 +T、CD19 +B、CD138 +B细胞水平的变化;CBA法检测血清炎症因子TNF-α、IL-17A、IL-6水平变化。 结果:预防用药及治疗用药组唾液流率( t=-5.81, P<0.001;t=-3.61, P<0.05)、浸润灶平均数目( t=3.95, P<0.01;t=4.94, P<0.001)、浸润灶平均面积( t=3.18, P<0.05;t=2.35, P<0.05)均具有明显改善。颌下腺组织中CD3 +CD4 +T( t=2.39, P<0.05;t=3.82, P<0.01)、CD44 +CD62L -CD4 +T( t=3.53, P<0.05;t=3.36, P<0.05)细胞明显下调。脾组织中CD3 +T( t=6.08, P<0.001;t=2.76, P<0.05)、CD3 +CD4 +T( t=3.73, P<0.05;t=2.39, P<0.05)、CD19 +B( t=5.88, P<0.001;t=4.23, P<0.01) 、CD138 +B( t=4.30, P<0.001;t=4.46, P<0.01)细胞也均明显下降。此外,用药组小鼠血清IL-17A水平均降低( t=4.15, P<0.01;t=3.36, P<0.01),预防用药组TNF-α水平下降( t=4.56, P<0.001),但IL-6水平变化无统计学意义。 结论:本研究提示来氟米特可预防、改善NOD鼠唾液腺功能减退及抑制体内免疫活化,为评估来氟米特对干燥综合征的治疗作用提供了理论依据。Objective To investigate the therapeutic effects of leflunomide on salivary gland secretion hypofunction in NOD mice with Sjogren′s syndrome.Methods NOD mice were randomly divided into four groups:prevention group,prevention control group,treatment group and treatment control group.Salivary flow rate was measured after pilocarpine stimulation.Changes in the average number and area of infiltrating lesions were compared after hematoxylin and eosin(HE)staining.The percentages of CD3+T,CD4+T,CD8+T,CD44+CD62L-CD4+T,CD19+B and CD138+B cells in submandibular gland and spleen were detected by flow cytometry.The levels of TNF-α,IL-17A and IL-6 in serum were detected by CBA method.Results The salivary flow rate(t=-5.81,P<0.001;t=-3.61,P<0.05),the number of infiltrating lesions(t=3.95,P<0.01;t=4.94,P<0.001)and the average area of infiltrating lesions(t=3.18,P<0.05;t=2.35,P<0.05)were significantly ameliorated in the prevention and treatment groups.Moreover,CD3+CD4+T cells(t=2.39,P<0.05;t=3.82,P<0.01)and CD44+CD62L-CD4+T cells(t=3.53,P<0.05;t=3.36,P<0.05)in the submandibular gland were significantly decreased.CD3+T(t=6.08,P<0.001;t=2.76,P<0.05),CD3+CD4+T(t=3.73,P<0.05;t=2.39,P<0.05),CD19+B(t=5.88,P<0.001;t=4.23,P<0.01)and CD138+B cells(t=4.30,P<0.001;t=4.46,P<0.01)in the spleen were also significantly reduced.Serum IL-17A(t=4.15,P<0.01;t=3.36,P<0.01)in the two groups and TNF-α(t=4.56,P<0.001)in the prevention group were down-regulated,but no significant difference was observed in IL-6 level.Conclusions This study suggested that leflunomide could prevent and improve salivary gland hypofunction and inhibit immune activation in NOD mice,providing reference for evaluating leflunomide in the treatment of Sjogren′s syndrome.

关 键 词：干燥综合征 来氟米特 NOD鼠 唾液腺功能 淋巴细胞 炎症因子 

分 类 号：R593[医药卫生—内科学]