摘要
目的 :观察小鼠烫伤后不同时相腹腔巨噬细胞的细胞膜及细胞质蛋白激酶C(PKC)的变化及应用特异性调控剂H 7后腹腔巨噬细胞内核因子 κB(NF κB)活性 ,从信号转导的角度阐明巨噬细胞功能紊乱的机制。方法 :以 15 %体表面积Ⅲ度烫伤小鼠为模型 ,收集腹腔巨噬细胞 ,同位素掺入法测膜浆PKC活性 ,电泳迁移率改变分析法检测NF κB的活性。结果 :烫伤后巨噬细胞细胞膜及细胞质PKC活性升高 ,其中胞膜PKC活性与TNF αmRNA的变化呈正相关。给予H 7后NF κB活性下降。结论 :烫伤后小鼠腹腔巨噬细胞PKC、NF κB的活性明显被激活 ,PKC NF κB信号通路参与了TNF α表达的调控。
Objective:To study the changes in the activity of protein kinase C(PKC)in murine peritoneal macrophages (PMФs) at different time intervals after severe scalding and to assay the activity of the nuclear factor κB (NF-κB) of PMФs under the action of H-7, a specific PKC inhibitor. These experiments were designed in an attempt to clarify the mechanisms underlying the functional disorders of macrophages from the angle of signal transduction. Methods:The experimental model was set up with mice which had been subjected to full-thickness grade Ⅲ scalding of 15% of the TBSA(total body surface area). From these animals, PMФs were collected. Both membrane and cytoplasmic PKC activities were assayed with the isotope(γ- 32P-ATP) incorporation method, while the NF-κB activity was detected with the electrophoretic mobility shift assay (EMSA). Results:Both membrane and cytoplasmic PKC activities were upregulated after scalding and the former showed a positive correlation with the changes in TNF-α mRNA. The NF-κB activity decreased under the action of H-7. Conclusions: Activity of both PKC and NF-κB was shown to be markedly increased following severe scalding. The PKC-NF-κB signal pathway seemed to be involved in the regulation of TNF-α expression in the macrophages.
出处
《医药导报》
CAS
2003年第6期367-369,共3页
Herald of Medicine
基金
全军九五课题<战创伤感染与免疫的研究>基金资助项目 (基金编号 :96L0 42 )
关键词
烫伤
巨噬细胞
核因子—κB
蛋白激酶C
Scald
Macrophages(MФs)
Nuclear factor-κB (NF-κB)
Protein kinase C(PKC)
