摘要
目的研究酪氨酸激酶抑制剂A77-1726对IL-13介导Dami细胞STAT6磷酸化和c-fos表达的影响,为A77-1726的临床应用和IL-13的信号通路研究提供新的实验依据。方法提取Dami细胞总RNA,RT-PCR检测c-fos mRNA表达。提取Dami细胞总蛋白,Western blot检测磷酸化STAT6和c-fos蛋白表达。凝胶定量软件Quantity One检测电泳条带光密度,统计学分析。结果100μg·L-1IL-13作用Dami细胞,可见STAT6磷酸化,50μmol·L-1A77-1726可阻断IL-13诱导的Dami细胞STAT6磷酸化。IL-13作用Dami细胞,c-fos mRNA表达增高(P<0.05),50μmol·L-1A77-1726阻断了IL-13诱导的Dami细胞c-fos mRNA的表达(P<0.05)。IL-13促进Dami细胞c-fos蛋白表达(P<0.05),50μmol·L-1A77-1726作用Dami细胞,阻断了IL-13诱导的c-fos蛋白表达(P<0.05)。结论酪氨酸激酶抑制剂A77-1726阻断了IL-13介导的白血病Dami细胞STAT6磷酸化和c-fos表达,JAK/STAT6通路是IL-13信号通路之一,IL-13诱导的c-fos表达与STAT6通路相关。
Aim To investigate the effects of tyrosine kinase inhibitor A77-1726 on IL-13 induced STAT6 phosphorylation and c-fos expression in Dami cell and to provide novel experimental basis to the clinical application of A77-1726 and the study of IL-13 pathway. Methods Total RNA was extracted from Dami cells incubated with or without IL-13 and A77-1726 respectively for various time points. RT-PCR and agar gel electrophoresis were used to examine the expression of c-fos mRNA. The expression of STAT6 and c-fos proteins was detected by Western blot. A densitometric relative quantitation of PCR and Western blot products was quantitated using Image Quant software. Results STAT6 was phosphorylated by treatment of 100 μg·L^-1 IL-13 in Dami cells. Phosphorylation of STAT6 induced by IL-13 was inhibited by treatment of 50μmol· L^-1 A77-1726. The expression of c-los mRNA was significantly induced by IL-13 treatment in Dami cells （P 〈0. 05 ）. c-los mRNA expression induced by IL-13 was inhibited by A77-1726 treatment （P 〈 0. 05 ）. The expression of c-los protein was induced by IL-13 treatment in Dami cells（P 〈0. 05） ,and IL-13 induced ex-pression of c-fos was transient. The expression of c-fos protein was also inhibited by A77-1726 treatment（P 〈 0. 05）. Conclusion IL-13 induced phosphorylation of STAT6 and c-fos expression was inhibited by tyrosine kinase inhibitor A77-1726. IL-13 activated JAK/STAT pathway. IL-13 induced c-fos expression was related with STAT6 pathway.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第7期906-910,共5页
Chinese Pharmacological Bulletin
基金
江西省自然科学基金资助项目(No0140036)
