维拉帕米减敏巨噬细胞机制研究  

Effects of Verapamil on expression of tyrosine phosphorylation and CD14 mRNA in peritoneal macrophages in burn mice

作　　者：贲道锋[1] 夏照帆[1] 刘志国[1] 杨勇[1] 郇京宁[1] 陈玉林[1] 

机构地区：[1]解放军第二军医大学长海医院烧伤科,上海200433

出　　处：《中国危重病急救医学》2000年第6期328-330,共3页Chinese Critical Care Medicine

基　　金：国家自然科学基金!资助项目 (39870 85 9);国家杰出青年科学基金!资助项目 (3972 5 0 2 9)

摘　　要：目的 :探讨维拉帕米对严重烧伤小鼠早期腹腔巨噬细胞 ( MΦ)内毒素 ( L PS)后信号转导途径的影响。方法 :BAL B/C小鼠随机分成烫伤组和假烫组 ,将收集到的腹腔 MΦ 加入不同浓度维拉帕米 ( 10、10 0和10 0 0 nmol/L )培养。用细胞免疫组化、逆转录聚合酶链反应 ( RT PCR)和酶联免疫方法 ,观察维拉帕米对小鼠 MΦ表达磷酸化酪氨酸 ( Tyr P)、CD14m RNA和产生肿瘤坏死因子α( TNFα)、白介素 6 ( IL 6 )的影响。结果 :与假烫组比较 ,烫伤后 2小时小鼠 MΦ Tyr P和 CD14表达明显增强 ,分泌细胞因子明显增加。维拉帕米各浓度对烫伤小鼠 MΦ CD14m RNA表达无明显影响 ;维拉帕米能使 MΦ 表达 Tyr P和分泌细胞因子TNFα、IL 6减少。结论 :维拉帕米减敏 MΦ 的机制可能与其影响 Tyr P的表达有关。Objective:To observe the effect of Verapamil on expression of tyrosine phosphorylation (TyrP) and CD14 mRNA,and secretion of cytokines in peritoneal macrophages from burn mice.Methods:Mice were randomly divided into burn group and sham group.Mice peritoneal macrophages from burn group ( n =10) or sham group ( n =10) were isolated and incubated with various doses of Verapamil (10,100,1 000 nmol/L). TyrP and CD14 mRNA expression in peritoneal macrophages were determined by immunohistochemistry and reverse transcription polymerase chain reaction.The cultured cell supernatant was collected to measure tumor necrosis factorα(TNFα) and interleukin6(IL6) levels by enzymelinked immunoassay.Results:Significant elevation of TyrP,CD14 expression and cytokine release were found at 2 hours postburn.Treatment with Verapamil in various doses didn′t markedly influence macrophage CD14 mRNA expression,while it could downregulate cytokine secretion and TyrP expression of macrophages in burn mice.Conclusions:Verapamil ( 1 000 nmol/L ) can downregulate cytokine secretion of macrophages in burn mice,which might be associated with inhibitory effect of TyrP expression.

关 键 词：维拉帕米 CD14 细胞因子 烧伤 小鼠 

分 类 号：R96[医药卫生—药理学]