摘要
目的 探讨烫伤大鼠伤后不同时间切痂对Th1细胞因子γ干扰素 (IFN γ)和Th2细胞因子白细胞介素 4 (IL 4 )循环水平的变化及其mRNA在脾脏T淋巴细胞表达的影响。方法 16 0只健康雄性Wistar大鼠背部 30 %Ⅲ度烫伤后 ,随机分为单纯烫伤对照组和伤后 8h、2 4h、96h切痂组 ,分别在伤后 4、12、2 4、4 8、96、12 0、16 8h活杀动物 ,采集血液和脾脏标本。ELISA法检测血浆内IFN γ、IL 4浓度 ;Ficoll分离液法、贴壁法和尼龙毛柱吸附法提取纯化脾脏T淋巴细胞 ,一步法提取细胞总RNA、RT PCR法扩增目的基因、琼脂糖凝胶电泳分析IFN γ和IL 4mRNA的表达。结果 烫伤 4h大鼠循环内上述细胞因子的浓度大幅升高 ,同时其mRNA表达迅速上调。Th1型细胞因子IFN γ及其mRNA表达在烫伤后 2 4h达峰值 ,其后逐渐下降 ;Th2型细胞因子IL 4的循环浓度及mRNA表达进行性增加 ;伤后 7d出现明显偏向Th2型反应的现象。切痂组两个细胞因子浓度增加及其mRNA表达上调的变化幅度小于单纯烫伤对照组 ,其中 8h切痂组的变化最小 ,2 4h和 96h切痂组次之。结论 休克期切痂有利于抑制严重烧伤后Th2型细胞因子的过度表达。
Objective To determine the serum level and mRNA expression of type-1/type-2 cytokines of T lymphocytes in spleens of rats after thermal injury and to investigate the effects of escharectomy during burn shock stage on IFN-γand IL-4 Methods One hundred and sixty male Wistar rats were randomized into four groups In group A, animals weren't subjected to escharectomy In groups B, C and D, escharectomy and skin allograft were performed at 8, 24, 96 hours postburn(PB) respectively At 4, 12, 24, 48, 96, 120 and 168 hours PB, animals were killed and blood and spleens samples were harvested ELISA was applied to determine the concentration of IFN-γand IL-4 in serum The expression pattern of IFN-γand IL-4 were observed at mRNA level in T lymphocytes isolated from spleen by RT-PCR. Results The serum levelof IFN-γand IL-4 rose rapidly and significantly after scald injury, expression of IFN-γand IL-4 mRNA in rats' T lymphocyte were also up-regulated spontaneously The serum level of IFN-γ and its mRNA expression began to rise within 4 hours PB, peaking at 24 hours PB Whereas IL-4 and its mRNA expression showed a persistent elevation Thereby leading to a dominant tendency of Th2 cytokine response on 7 d PB In group A all above parameters revealed most obvious changes compared with controls, then ranked in group D, B and C Conclusion Escharectomy during burn shock stage is helpful to decrease the harmful over expression of Th2-type lymphocyte after severe thermal injury
出处
《中华外科杂志》
CAS
CSCD
北大核心
2004年第18期1142-1145,共4页
Chinese Journal of Surgery
