CYP-EET信号通路对大鼠MCAO缺血再灌注后血管新生影响的研究  

Effects of CYP-EET signaling pathway on angiogenesis after MCAO ischemia-reperfusion in rats

作　　者：刘阳 郑小龙 喻志源[1] 王伟[1] 谢敏杰[1] Liu Yang;Zheng Xiaolong;Yu Zhiyuan;Wang Wei;Xie Minjie(Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China)

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《疑难病杂志》2020年第5期500-504,515,共6页Chinese Journal of Difficult and Complicated Cases

基　　金：国家自然科学基金资助项目(81501020,81371312);湖北省自然科学基金资助项目(2019CFB678)。

摘　　要：目的研究CYP-EET信号系统对大鼠大脑中动脉闭塞(MCAO)缺血再灌注后血管新生及VEGF/VEGFR信号通路的影响。方法2017年6月—2018年4月于华中科技大学同济医学院附属同济医院进行实验。选择SPF级健康成年SD雄性大鼠80只,建立大鼠MCAO缺血再灌注模型,采用ELISA法检测脑缺血后1、3、7 d脑组织内14,15-EET的变化;通过侧脑室注射14,15-EET或sEH选择性抑制剂AUDA增加内源性EETs的含量,应用免疫荧光技术和Western-blot检测脑缺血再灌注1、3、7 d后血管新生及相关信号通路。结果与假手术组比较,缺血组脑缺血再灌注后1 d脑组织内14,15-EET含量显著降低(t/P=11.863/0.000),脑缺血再灌注后3、7 d脑内14,15-EET含量显著增加(t/P=-10.411/0.000,-8.432/0.000);外源性加入14,15-EET或AUDA后vWF、VEGFR、VEGF表达增加(F/P=71.122/0.000、44.363/0.000、45.034/0.000),p-Akt及p-Erk蛋白表达降低(F/P=39.601/0.000、121.670/0.000)。结论脑缺血再灌注病理过程中CYP-EETs信号系统通过上调VEGF/VEGFR通路,抑制Erk及Akt磷酸化,促进血管新生。Objective To study the effect of CYP-EET signaling system on angiogenesis and VEGF/VEGFR signaling pathway after ischemia-reperfusion in rats with middle cerebral artery occlusion(MCAO).Methods From June 2017 to April 2018,the experiment was conducted in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.The MCAO ischemia-reperfusion model was established in 80 SPF healthy adult SD male rats.The changes of 14,15-EET in brain tissue were detected by ELISA on the 1st,3rd and 7th day after cerebral ischemia.The content of endogenous EETs was increased by injecting 14,15-EET or the selective inhibitor AUDA into the lateral ventricle.Neovascularization and related signal pathways were detected by immunofluorescence and Western blot after 1,3,7 days of cerebral ischemia-reperfusion.Results Compared with the sham operation group,the 14,15-EET content in the brain tissue of the ischemia group decreased significantly(t/P=11.863/0.000),and the 14,15-EET content in the brain increased significantly(t/P=-10.411/0.000,t/P=-8.432/0.000)on the 3rd and 7th day after cerebral ischemia-reperfusion.The expression of vWF,VEGFR and VEGF increased(F/P=71.122/0.000,F/P=44.363/0.000,F/P=45.034/0.000),and the expression of p Akt and p Erk decreased(F/P=39.601/0.000,F/P=121.670/0.000)after exogenous addition of 14,15-EET or AUDA.Conclusion CYP-EETs signal system can inhibit ERK and Akt phosphorylation and promote angiogenesis by up regulating VEGF/VEGFR pathway during cerebral ischemia-reperfusion.

关 键 词：CYP-EETs信号系统 脑缺血再灌注 血管新生 神经保护 大鼠 

分 类 号：R73[医药卫生—肿瘤]