EPO通过AMPK-KLF2信号通路调节脑缺血后血管新生的分子机制研究  

Molecular mechanism of EPO regulating angiogenesis after cerebral ischemia via AMPK-KLF2 signaling pathway

作　　者：倪广晓[1] 王璞[2] 段春巧 孟然[1] 周宏斌[1] Ni Guangxiao;Wang Pu;Duan Chunqiao;Meng Ran;Zhou Hongbin(First Department of Physical Medicine and Rehabilitation,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Prosthodontics,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院康复医学一科,石家庄050000 [2]河北医科大学第二医院口腔修复科,石家庄050000

出　　处：《广西医科大学学报》2020年第2期218-223,共6页Journal of Guangxi Medical University

摘　　要：目的:探讨促红细胞生成素(EPO)通过腺苷酸活化蛋白激酶-Kruppel样转录因子2(AMPK-KLF2)信号通路调节脑缺血后血管新生的机制。方法:选取60只健康雄性C57BL/6小鼠,随机分为假手术(Sham)组、大脑中动脉闭塞缺血(MCAO)组和MCAO+EPO处理组,每组20只。采用改良ZeaLonga法制备小鼠MCAO模型,MCAO+EPO处理组小鼠在再灌注即刻腹腔注入EPO(5000 IU/kg),术后隔天注射1次。记录小鼠的死亡数量及对小鼠进行神经功能评分。在脑缺血后第7天,提取脑组织蛋白,采用Western blot检测脑组织中EPO、血管内皮生长因子(VEGF)、血管内皮生长因子受体(KDR)、AMPK、低氧诱导因子-1α(HIF-1α)、KLF2和一氧化氮合酶(eNOS)蛋白表达。结果:与MCAO组相比,MCAO+EPO组小鼠生存率明显提高,神经功能明显改善(P<0.01)。与Sham组相比,MCAO组小鼠脑组织中EPO、HIF-1α蛋白表达明显增高,VEGF、KDR、AMPK、KLF2和eNOS蛋白表达明显降低(均P<0.01);与MCAO组相比,MCAO+EPO处理组小鼠脑组织中EPO、VEGF、KDR、AMPK、KLF2和eNOS蛋白含量明显增高,HIF-1α蛋白表达明显降低(均P<0.01)。结论:EPO能够通过激活AMPK-KLF2信号通路,促进VEGF及其受体KDR的表达,参与调控HIF-1α和eNOS蛋白表达,从而在脑缺血后促进血管新生。Objective:To investigate the mechanism of erythropoietin(EPO)regulating angiogenesis after cerebral ischemia via adenosine monophosphate-activated protein kinase-Krüppel-like factor 2(AMPK-KLF2)signaling pathway.Methods:Sixty healthy male C57BL/6 mice were randomly divided into sham group,middle cerebral artery occlusion(MCAO)group and MCAO+EPO treatment group,with twenty mice in each group.MCAO models of mice were prepared by the modified ZeaLonga method.In the MCAO+EPO treatment group,mice were received intraperitoneal injection of EPO(5000 IU/kg)immediately after reperfusion and once every other day afterwards.The number of dead mice and the scores in neurological function were recorded.On the 7th day after surgery,brain tissue proteins were extracted,and western blot was used to detect the protein expression of EPO,vascular endothelial growth factor(VEGF),kinase insert domain receptor(KDR),AMPK,hypoxia-inducible factor-1α(HIF-1α),KLF2,and endothelial nitric oxide synthase(eNOS).Results:Compared with the MCAO group,the survival rate and the neural function were significantly improved of the mice in the MCAO+EPO group(P<0.01).Compared with the sham group,the protein expression of EPO and HIF-1αwere significantly increased,and the protein expression of VEGF,KDR,AMPK,KLF2e,and NOS were significantly reduced(all P<0.01)in the brain tissue of MCAO mice.Compared with the MCAO group,the protein expression of EPO,VEGF,KDR,AMPK,KLF2,and eNOS were significantly increased,whereas the protein expression of HIF-1α was decreased significantly(all P<0.01)in the brain tissue of the MCAO+EPO-treated mice.Conclusion:EPO can promote the expression of VEGF and its receptor KDR by activating the AMPK-KLF2 signaling pathway,participate in the regulation of HIF-1α and eNOS protein expression,and thereby promote angiogenesis after cerebral ischemia.

关 键 词：促红细胞生成素 AMPK KLF-2 血管新生 

分 类 号：R965[医药卫生—药理学]