转染TGF-β1基因的树突状细胞对重症肌无力大鼠T细胞IFN-γ和NO分泌的调节  

Regulatory effect of dendritic cells transfected with TGF-β1 gene on secretion of IFN-γ and NO of T lymphocytic cells in rats with experimental autoimmune myasthenia gravis

作　　者：王云甫[1] 陈吉相[2] 孙圣刚[2] 曹学兵[2] 

机构地区：[1]郧阳医学院附属太和医院神经内科,湖北十堰442000 [2]华中科技大学同济医学院附属协和医院神经科,湖北武汉430022

出　　处：《第四军医大学学报》2009年第18期1662-1664,共3页Journal of the Fourth Military Medical University

基　　金：湖北省自然科学基金(2006ABA344);湖北省教育厅科学研究项目(2004Z001)

摘　　要：目的:探讨TGF-β1基因转染的树突状细胞(TGF-β1-DC)对实验性自身免疫性重症肌无力(EAMG)大鼠T细胞IFN-γ和NO分泌的调节作用.方法:近交系、8～10 wk龄、雌性Lewis大鼠25只,随机分为5组:正常组、EAMG组、DC对照组、TGF-β1-DC治疗组、生理盐水治疗对照组.除正常组外,其余各组均采用丁氏双鳍电鳐的电器官乙酰胆碱受体蛋白二次免疫的方法复制EAMG大鼠模型.初次免疫后第5日分别皮下注射2×106的DC,TGF-β1-DC及等体积的生理盐水,EAMG组不接受任何治疗.初次免疫后7 wk,分离脾脏T细胞体外培养48 h后分别应用ELISA和化学方法检测T细胞IFN-γ和NO的分泌水平.结果:正常大鼠T细胞体外培养48 h后,其上清液中IFN-γ的含量很少,而EAMG组大鼠T细胞培养上清液中IFN-γ的含量显著增加(P<0.01).TGF-β1-DC治疗组、DC对照组、生理盐水对照组IFN-γ的含量与EAMG组比较均无统计学差异;正常大鼠T细胞体外培养的上清液中NO的含量很少,EAMG组大鼠T细胞培养上清液中NO的含量明显增加(P<0.01).TGF-β1-DC治疗组NO的含量较EAMG组明显增加(P<0.05),DC对照组、生理盐水对照组与EAMG组比较无统计学差异.结论:调节EAMG大鼠体内IFN-γ和NO的分泌可能是TGF-β1-DC的治疗机制之一.AIM：To explore the regulatory effect of dendritic cells （DC） transfected with TGF-β1 gene on the secretion of IFN- γ and NO of T lymphocytic cells in the Lewis rats with experimental autoimmune myasthenia gravis （EAMG）. METHODS： Twenty-five healthy female Lewis rats were divided randomly into 5 groups： normal group, EAMG group, DC treatment group, peDNA3-TGFβ1-DC treatment group and normal saline group. All the rats except those in normal group were immunized with aeetylcholine receptor （AChR） protein extracted from electric organ of Narcine timilei and completed Freund＇s adjuvant （CFA）. 2 × 10^6 pcDNA3-TGFβ1-DCs/rat were injected subcutaneously into the hacks of the rats immunized 5 d earlier with AChR ＋ CFA. The rats in DC treatment group and normal saline group were injected with untreated DCs and normal saline respectively. Seven weeks after the first immunization, T lymphocytic cells in every rat were separated from the spleen. After a 48-h culture, the content of IFN-γ in the supernatant was detected by ELISA and the level of NO on the T lymphocytic cells was examined using chemistry method. RESULTS ： Low content of IFN-γ in the supernatant was found in the normal rats and the content increased markedly in EAMG rats （ P 〈0.01 ）. No significant difference was observed in the contents of IFN-γ between EAMG group and pcDNA3-TGF- β1-DC group. The level of NO was low in normal rats but increased significantly in EAMG rats （ P 〈 0. 001 ）. The level in pcDNA3-TGFβ1-DC group was higher than that in EAMG group （ P 〈 0.05 ）, but with no statistically difference between EAMG group, DC treatment group and normal saline group. CONCLUSION ： The regulation of the excretion of IFN-γ and NO on T lymphocytic cells may play a critical role in the treatment with DCs transfected with pcDNA3-TGFβ1 in EAMG rats.

关 键 词：重症肌无力 转化生长因子 树突状细胞 基因转染 干扰素-Γ 一氧化氮 

分 类 号：R746.1[医药卫生—神经病学与精神病学]