摘要
目的探讨缺氧对钙／钙调蛋白依赖性丝氨酸蛋白激酶（CASK）表达的影响及其与p38MAPK信号通路活化的关系。方法采用定量PCR方法Western blot技术，检测内皮细胞株的CASK在常氧和缺氧1、3、6、12h条件下表达情况。采用Western blot技术，检测常氧和缺氧1、3、6、12h条件下p38 MAPK 180位苏氨酸／182位酪氨酸双位点磷酸化情况，及p38信号通路特异性阻断剂SB203580预处理1h，血管内皮细胞缺氧3h后CASK蛋白的表达。采用双荧光素酶报告系统，分析常氧和缺氧1、3、6．12h条件下CASK启动子报告基因的活性。结果缺氧能够明显上调CASK mRNA和蛋白的表达，缺氧3h，CASK mRNA的表达为常氧对照组的1．8倍，CASK蛋白表达为常氧对照组的1．4倍。p38MAPK信号通路阻断剂SB203580预处理，能够以剂量依赖性方式抑制3h缺氧诱导的内皮细胞CASK蛋白表达。不同时间的缺氧能够明显增强CASK荧光素酶报告基因的活性。结论缺氧能够诱导内皮细胞CASK的表达部分依赖于p38MAPK信号的活化。
Objective To examine the expression of calcium/calmodulin-dependent serine protein kinase （CASK） triggered by hypoxia and explore the role of p38 mitogen activated protein kinase （p38 MAPK） in this induction. Methods The expression of CASK mRNA and protein under normoxia and hypoxia （1st,3rd,6th and 12th h） was detected by using real time RT-PCR and Western blot respectively, and the changes of the CASK expression under the condition of p38 MAPK inhibitor SB203580 pretreatment （ 1st h） were observed. The effects of hypoxia on CASK promoter activity were investigated by u- sing dual luciferase reporter system. Results Hypoxia could induce the CASK mRNA and protein expression. CASK mRNA expression was 1.8 folds and CASK protein expression 1.4 folds to normoxia at 3rd h after hypoxia, and p38 MAPK specific inhibitor, SB203580 could decrease the CASK protein expression in a dose-dependent manner. Hypoxia could elevate the CASK promoter activity. Conclusion Short-term hypoxia-induced the CASK expression is p38 MAPK dependent.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第1期39-41,共3页
Chinese Journal of Experimental Surgery
基金
国家重点基础研究发展计划（2005CB522601）
国家自然科学基金重点资助项目（10332060）
