高迁移率族蛋白B1对烫伤大鼠脾脏调节性T细胞免疫功能的影响  

Effect of high mobility group box-1 protein on immunsuppression function of splenic regulatory T cells in rats with thermal injury

作　　者：黄立锋[1] 姚咏明[1] 张立天[1] 董宁[1] 于燕[1] 盛志勇[1] 

机构地区：[1]解放军总医院第一附属医院全军烧伤研究所基础部,北京100048

出　　处：《中华创伤杂志》2009年第10期936-940,共5页Chinese Journal of Trauma

基　　金：基金项目：国家重点基础研究发展规划资助项目（2005CB522602）;国家自然科学基金资助项目（30672178,30872683）;国家杰出青年科学基金资助项目（30125020）

摘　　要：目的观察严重烫伤延迟复苏大鼠体内高迁移率族蛋白B1（HMGBI）的变化及其对调节性T细胞（Treg）免疫功能的影响。方法采用30％体表面积Ⅲ度烫伤延迟复苏模型，将136只大鼠按随机数字表法分为正常对照组（8只）、假烫伤组（32只）、烫伤组（32只）、丙酮酸乙酯（EP）干预组（32只）及晚期糖基化终末产物受体（RAGE）抗体干预组（32只）。后4组实验动物再分为4个亚组，分别于烫伤后1，3，5，7d无菌取血和脾脏，免疫磁珠法分离大鼠脾脏CD4^＋ CD25^＋Treg。采用ELISA检测血清HMGB1水平，应用流式细胞术检测Treg表面晚期糖基化终末产物受体（RAGE）、细胞毒性T淋巴相关抗原4（CTLA-4）及又头翼状螺旋转录因子P3（Foxp3）表达。结果（1）烫伤延迟复苏导致大鼠血清HMGB1水平在伤后1—7d显著升高，于第3天达峰值（P〈0．01）。EP干预后烫伤大鼠血清HMGB1水平1～7d均明显降低。RAGE抗体干预对烫伤大鼠血清HMGB1水平无明显影响（P〉0．05）。（2）与假烫伤组比较，严重烫伤后1～7d脾脏Treg表面RAGE、CTLA-4及Foxp3表达均不同程度增强。EP干预组及RAGE抗体干预组上述三项指标较烫伤组明显降低（P〈0．05或0．01）。结论严重烫伤后HMGB1可通过RAGE受体对Treg免疫抑制活性产生影响，进而参与烫伤后脓毒症的发病过程。Objective To observe the changes of high mobility group box-1 protein （ HMGB1 ） and its effects on immunosuppressin function of splenic regulatory T cells （Treg） in rats with thermal injury. Methods A total of 136 Wistar rats were subjected to 30% full-thickness scald injury followed by delayed resuscitation. All the rats were randomly divided into normal control group （ n = 8 ） , sham burn group （ n = 32）, burn group （ n ： 32）, ethyl pyrnvate （EP） treatment group （ n = 32） and anti-receptor for advanced glycation end product （ RAGE ） antibody treatment group （ n ： 32 ）. Four groups were further divided into four subgroups （eight rats per group） , which were sacrificed on post burn days 1, 3, 5 and 7, respectively, and blood as well as spleen samples were harvested aseptically. CD4^＋ CD25^＋ Treg was isolated from the spleens by immunomagnetic beads. Serum HMGB1 levels were determined by ELISA, and the expressions of RAGE, eytotoxic T lymphocyte-associated antigen 4 （CTLA-4） and fork- head/winged helix transcription factor 3 （ Foxp3 ） were detected by flow cytometry. Results （ 1 ） The expression of serum HMGB1 was significantly elevated on post burn days 1-7 and peaked on post burn day 3 （P 〈0.01 ）. The expression of serum HMGB1 was significantly inhibited by EP, but not by anti-RAGE antibody （P 〉0.05）. （2） Compared with sham burn group, the expressions of RAGE, CTLA-4 and Foxp3were increased in rats with thermal injury on post burn days 1-7. The expressions of RAGE, CTLA-4 and Foxp3 in EP treatment group and anti-RAGE antibody treatment group were significantly decreased （P 〈 0.05 or P 〈0.01 ）. Conclusions HMGBI influences post burn immunosuppression of splenic CD4^＋ CD25^＋ Treg via binding RAGE on the surface of Treg, thereby involves in the development of immune dysfunction in sepsis following severe thermal injury.

关 键 词：创伤和损伤 高迁移率族蛋白质类 T淋巴细胞 调节性 翼螺旋转录因子 类 细胞毒性T淋巴相关抗原4 

分 类 号：R644[医药卫生—外科学]