门静脉注射髓鞘碱性蛋白诱导免疫耐受的研究  

An experiment study on the induction of immune tolerance by the injection of myelin basic protein in hepatic portal vein

作　　者：刘勇[1] 亢建民[2] 张明超[2] 崔占群 杨伟健[2] 闫华[2,3] 

机构地区：[1]天津医科大学研究生院,300070 [2]天津市环湖医院神经外科,300060 [3]天津市脑血管与神经变性重点实验室,300060

出　　处：《中华实验外科杂志》2016年第4期975-978,共4页Chinese Journal of Experimental Surgery

基　　金：天津市自然科学基金（13JCYBJC21100）;天津市科技支撑计划（13ZCZDSY01600）

摘　　要：目的观察门静脉注射同种脑髓鞘碱性蛋白（MBP）诱导免疫耐受及肝脏Kupffer细胞在免疫耐受建立中的作用。方法将32只雄性sD大鼠随机分为4组，每组8只，A组：假手术组；B组：生理盐水对照组；C组：MBP实验组；D组：氯化钆（GdCl3）预处理组（门静脉注射前24h经鼠尾静脉注射GdCl3溶液）。造模后10d鼠尾静脉注射MBP致敏，致敏后24h和48h分别行酶联免疫吸附试验法（ELISA）检测外周血IBP抗体浓度、转化生长因子-β1（TGF-β1）浓度；流式细胞术检测外周血CD4^±／CD8^±T细胞比值；鼠尾静脉注射MBP致敏后48h处死大鼠，实时定量聚合酶链反应（Real—timePCR）检测肝组织Fas配体（FasL）表达含量。结果与A、B、D组比较，C组致敏后24．48h外周血MBP抗体浓度显著降低（P〈0．05），各组浓度分别为[（78．63±9．64）、（76．13±7．57）、（49．14±8．51）、（68．71±11．56）比（82．00±5．71）、（79．00±6．47）、（47．00±5．28）、（75．00±4．96）ng／m]]、外周血CD4^±／CD8^±T细胞比值降低（P〈0．05），24h表达量分别为（1．94±0．31、2．00±0．14、1．46±0．22、1．93±0．14）；48h表达量分别为（1．96±0．09、2．04±0．19、1．56±0．26、2．01±0．18）；C组致敏后48h肝组织FasL表达含量升高（P〈0．05），各组表达含量（×10^-4）分别为（5．72±2．78、3．77±4．76、17．44±13．92、9．38±5．14）。与A、B组比较，C组致敏后24、48h外周血TGF—β1浓度升高（P〈0．05）；D组致敏后24、48h外周血TGF-β1浓度升高（P〈0．05）；D组与C组比较，差异无统计学意义（P〉0．05），各组浓度分别为[（24．05±4．96）、（24．74±1．98）、（37．51±6．12）、（31．64±6．20）比（23．11±5．26）、（22．57±2．85）、（34．76±4．88）、（30．10±7．17）ng／L]。结论门静脉注射MBP可诱导机体免疫耐受的建立，可降低免�Objective To explore the induction of immune tolerance by the injection of myehn basic protein （MBP） into hepatic portal vein and the role of Kupffer cells in this process. Methods 32 male SD rats were randomly divided into four groups, 8 in each group： A group： sham group, B group： normal saline control group, C group： MBP experiment group, D group： GdCI3 pre treatment group （injec- ted GdCI3 into rat tail vein 24 h before the portal vein injection）. Inject MBP into rat tail vein 10 days after the portal vein injection. MBP antibody and transforming growth factor - β1 （ TGF - β1 ） were detected by enzyme linked immunosorbent assay （ ELISA）, CD4^＋CD8^＋T cells ratio in peripheral blood detected by flow cytometry （FCM） 24 h and 48 h after the rat tail vein injection. 48 h after rat tail vein injection, Fas ligand （FasL） was measured by real -time quantitative polymerase chain reaction （Real- time PCR）.Results Compared with group A, B and D, the concentration of MBP antibody were extraordinarily lower in group C （P 〈0. 05）, concentration respectively were [（78. 63 ±9. 64）, （76. 13 ±7. 57）, （49. 14 ± 8.51）, （68.71±11.56） vs. （82.00±5.71）, （79.00±6.47）, （47.00±5.28）, （75.00± 4. 96） ng/ml ], meanwhile CD4^＋/CD8^＋ T ratio were obviously lower in group C （ P 〈 0. 05 ） at 24 h and 48 h after rat tail vein injection, the ratio were （ 1.94 ± 0. 31, 2. 00 ± 0. 14, 1.46 ± 0. 22, 1.93 ± 0. 14 ; 1.96 ±0. 09, 2.04 ±0. 19, 1.56 ±0. 26, 2. Ol ±0. 18）, the gene expression of FasL were notably higher than that in group C （P 〈 0. 05 ） at 48 h after rat tail vein injection, concentration （ × 10^-4） respectively were （5.72 ± 2.78, 3.77 ± 4. 76, 17.44 ± 13.92, 9. 38 ± 5.14）. Compared with group A and B, the concentration of TGF - β1 were obviously higher than that in group C and D （ P 〈 0. 05 ） and there were no difference between group C and D at 24 h and 48 h after rat tail vein injection, concentra

关 键 词：免疫耐受 门静脉 脑髓鞘碱性蛋白 KUPFFER细胞 

分 类 号：R654.2[医药卫生—外科学]