B10细胞在益气升提法治疗EAMG大鼠中的作用  

Effects of B10 cells in EAMG Rats Treated by the Method of Invigorating Qi for Ascending

作　　者：许梦贤 尚卫兵 魏亚男 姬梦姣 吴颢昕[1] Xu Mengxian;Shang Weibing;Wei Yannan;Ji Mengjiao;Wu Haoxin(Basic Medical College,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学基础医学院,南京210023

出　　处：《世界科学技术-中医药现代化》2020年第1期157-163,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：江苏省中医药管理局全国名老中医药专家传承工作室建设项目(013007001009):孟景春全国名老中医药专家传承工作室,负责人:吴颢昕.

摘　　要：目的观察在实验性自身免疫性重症肌无力(experimental autoimmune myasthenia gravis,EAMG)中,用益气升提法对其治疗的效果及分析B10细胞的作用和免疫机制。方法将48只Lewis大鼠随机分成6组:阳性药物对照组、益气升提低剂量组、益气升提中剂量组、益气升提高剂量组、佐剂对照组和模型对照组。用Rα97-116免疫大鼠建立EAMG模型。在实验中对大鼠体重,临床表现,低频重复电刺激(RNS)衰减率进行观察。检测血清中乙酰胆碱受体抗体(AchR-Ab)、白介素10(IL-10)的含量,其方法是酶联免疫吸附试验(ELISA);检测组织中CD19的表达,其方法用蛋白印迹法(western-blot),以及用逆转录-聚合酶链反应(RT-PCR)检测IL-10和CD19基因的表达。结果给药后,大鼠体重回升,临床症状好转,RNS衰减率显著降低(P <0.05,P <0.01),血清中AchR-Ab含量显著下降(P <0.05,P <0.01),IL-10含量显著上升(P <0.05,P <0.01),组织中CD19蛋白表达下降(P <0.05,P <0.01)。CD19和IL-10基因表达下降(P <0.01,P <0.001)。结论 EAMG大鼠经过益气升提法治疗后,临床症状及体重下降趋势有明显改善作用,其机制可能是升高IL-10含量,下调IL-10和CD19基因表达以及下调CD19蛋白表达,降低血清中AchR-Ab含量,从而减少神经肌肉接头处AchR损害。Objective To observe the role of B10 cells in experimental autoimmune myasthenia gravis(EAMG)rats treated by the method of invigorating qi for ascending and investigate the immune mechanism.Methods 48 Lewis rats were randomly divided into 6 groups:positive control group,invigorating qi for ascending astragalus low dose group,invigorating qi for ascending astragalus middle dose group,invigorating qi for ascending astragalus high dose group,CFA group and EAMG model group.The rats were immunized with Rα97-116.Evaluate rats’weight changes and clinical symptoms and the decrement percentage of RNS.AchR-Ab and IL-10 levels was detected by ELISA in serum.WB was adopted to determine the expression of CD19 protein in tissues.RT-PCR was adopted to determine the expression il-10 and CD19 genes in tissues.Results After medication,the percentage of decrement of RNS of rats in invigorating qi for ascending and positive control groups were decreased significantly(P<0.05,P<0.01),and rats’weight and other symptoms were improved significantly(P<0.001);the titers of AChR-Ab was decreased significantly(P<0.05,P<0.01),with the titer of IL-10 increased significantly(P<0.01);The expression of CD19 protein was down-regulated(P<0.01,P<0.05).The expression of CD19 and IL-10 gene was down-regulated(P<0.01,P<0.001).Conclusion invigorating qi for ascending groups can improve body weight and symptoms of EAMG rats.The immune mechanism of treatment may be promot IL-10 content and down-regulate the expression of CD19 protein and CD19 and IL-10 gene,inhibiting the synthesis of AchR-ab so that the damage of AchR of NMJ will be reduced.

关 键 词：益气升提法 实验性自身免疫性重症肌无力 B淋巴细胞亚群 实时荧光定量-聚合酶链反应 

分 类 号：R2-031[医药卫生—中西医结合]