摘要
目的 :探讨烧伤血清对离体大鼠肺泡巨噬细胞 (AM )CD14的mRNA基因和膜蛋白 (mCD14 )表达变化及其对AM分泌细胞因子的影响。 方法 :分离并收集大鼠AM ,以烧伤血清及LPS刺激 ,再分别以抗CD14抗体作用后提取总RNA ,用RT PCR方法检测不同时相点CD14mRNA表达 ;用ELISA方法检测培养液中TNF α和IL 6浓度 ;免疫组化法检测AM膜CD14蛋白表达变化。 结果 :经烧伤血清和LPS刺激后 ,AM的CD14基因以及蛋白表达从 1h起就开始增加 ,2h达峰值 ,然后逐渐减弱。上述改变在伤后 12h内持续 ,细胞因子分泌相应增加 ;抗CD14抗体阻断CD14作用后 ,CD14的基因和蛋白表达显著降低 ,细胞因子分泌亦相应减少。 结论 :严重烧伤后可能随着LPS增加 ,通过激活内毒素信号传导通路 ,使AM分泌细胞因子增加。这种作用可以被抗CD14抗体所阻断 。
Objectives: To observe the AM CD14 mRNA and mCD14 protein expression and the cytokine(TNF α?IL 6) concentration of severely burned rat in early stage. Methods: SD rat were burned 20% TBSA Ⅲ injury . The AM isolated at 1,2,3,4,6,8,12 h after burn. The obtained AM to assay CD14 mRNA and mCD14 protein expression by RT PCR and immunohistochemical method after 30% burn serum effected. TNF αand IL 6 concentration of the culture liquid assay at the same time. Results: CD14 mRNA and mCD14 expression was found to increase significantly in alveolus Macrophages. These effects peaked at 2 h and lasted to 12 h.At the same time,the TNF α,IL 6 concentration of culture liquid elevated correspondingly.When CD14 antibody used,the expression of CD14 mRNA and mCD14 decreased significantly.The TNF α,IL 6 concentration decreased at the same time. Conclusions: AM CD14 mRNA and mCD14 expression was upregulated after burn. Perhaps this change related with the increased concentration of LPS after severely burn. Such Regulation may be blocked by the CD14 antibody and decrease the TNF αand IL 6 concentration.
出处
《医学研究生学报》
CAS
2002年第6期478-481,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目 (批准号 :3 9870 85 9)
