严重烫伤后大鼠外周血中性粒细胞和淋巴细胞TNF-α的表达

TNF-α mRNA expression in the peripheral blood neutrophils and lymphocytes of severely burned rats

目的 研究大鼠烫伤后外周血中性粒细胞和淋巴细胞的TNF αmRNA表达的动态变化 ,以了解严重烫伤对外周血中性粒细胞和淋巴细胞表达TNF α的影响。方法 复制 3 0 %体表面积Ⅲ度烫伤大鼠模型 ;分离外周血中性粒细胞和淋巴细胞 ;提取总RNA ,采用RT PCR分析TNF αmRNA的表达。结果 PMN中TNF αmRNA的表达在伤后的 6～ 2 4h内较伤前明显上调 ,而淋巴细胞在伤前未发现TNF αmRNA的明显表达 ,但伤后 3～ 48h表达急剧上升 ;两者均于伤后 12h达高峰 ,以后缓慢下降 ,至 48h仍维持较高水平。结论 大鼠烫伤早期外周血中性粒细胞和淋巴细胞TNF αmRNA的表达明显上调 ,表明严重烫伤可显著影响血中性粒细胞和淋巴细胞TNF

Objective To study the dynamic changes of TNF α mRNA expression in the peripheral blood neutrophils and lymphocytes in postburn rats. Methods Wistar rats inflicted with 30% total body surface area (TBSA) Ⅲ degree scald were employed as the model. The peripheral blood neutrophils and lymphocytes were isolated by density gradient centrifugation using Percoll hypaque and Ficoll hypaque, respectively. Total RNA of the cells was obtained and purified with Tripure Isolation Reagent. The TNF α mRNA expression was analyzed by Titan One Tube RT PCR Kit. Results The up regulation of TNF α mRNA expression was found in peripheral blood neutrophils and lymphocytes at 6, 12, 24, 48 h and at 3, 6, 12, 24 and 48 h after burn, respectively. TNF α mRNA expression in both cells reached the peak at 12 h after burn and decreased gradually but remained at a high level at 48 h after burn. Conclusion The up regulation of TNF α mRNA expression in peripheral blood neutrophils and lymphocytes of postburn rats suggests that severe burn injury may have significant effect on TNF αexpression in peripheral blood neutrophils and lymphocytes.