脑血管内皮细胞Prmt5在脑血管发育过程中的表达及其下游靶分子  

Expression of Prmt5 in Cerebrovascular Endothelial Cells and its Downstream Targets During Cerebrovascular Development

作　　者：梁爽 杨晓 王俊 LIANG Shuang;YANG Xiao;WANG Jun(State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences(Bei.jing),Beijing Institute of Lifeomics,Beijing 102206,China)

机构地区：[1]军事医学研究院生命组学研究所,北京102206

出　　处：《生物技术通讯》2019年第2期147-152,共6页Letters in Biotechnology

基　　金：国家自然科学基金重点项目(31430057)

摘　　要：目的:研究内皮细胞中蛋白精氨酸甲基转移酶5(PRMT5)在脑血管发育及血脑屏障建成的关键时期的表达变化及其潜在下游靶分子。方法:原位观察PRMT5在小鼠不同发育时期脑血管内皮上的分布;流式分选获得原代脑血管内皮,利用real-time PCR分析Prmt5的表达;体外培养小鼠内皮细胞敲降Prmt5后,利用Western印迹、real-time PCR、ChIP等方法检测其对经典下游分子的影响。结果:PRMT5在胚胎期各时间点的脑血管内皮细胞质均有表达,小鼠出生后主要表达在脑血管内皮细胞核,在胚胎期18.5 d时表达量显著升高;小鼠脑血管内皮细胞体外细胞系中基因敲降Prmt5后,其经典对称甲基化组蛋白产物H4R3me2s及H3R2me2s均明显下降,Bmp4表达显著上调;免疫共沉淀实验提示Bmp4启动子区域组蛋白具有H3R2me2s修饰,Prmt5基因敲降后,该组蛋白修饰显著减少。结论:脑血管发育过程中PRMT5在脑血管内皮细胞中表达的位置和水平均发生变化,脑血管内皮细胞中PRMT5可以调节H4R3和H3R2对称二甲基化水平,Bmp4启动子区域组蛋白具有H3R2me2s修饰,且PRMT5可以抑制Bmp4表达。Objective:To analyze the distribution of protein arginine methyltransferases 5(PRMT5)in brain endothelial cells during the cerebral vascular development,and to identify the potential target genes of Prmt5.Methods:The distribution of PRMT5 on cerebral vascular endothelium was observed in situ.Primary cerebral vascular endothelial cells were obtained by flow sorting,and real-time PCR was used to analyze the dynamic expression of Prmt5.Western blotting,real-time PCR and ChIP assay were used to identify the downstream signaling of Prmt5.Results:PRMT5 was expressed in the both cytoplasm and nucleus of cerebrovascular endothelial cells during the embryonic stage.However,the expression of Prmt5 restricted in the nucleus of brain endothelial cells after birth.The expression level of Prmt5 in E18.5 was significantly higher.Furthermore,we found that H4R3me2s and H3R2me2s were down-regulated in bEnd3 after siPrmt5 treatment.Expression of Bmp4 was inhibited by Prmt5.The histones in the Bmp4 promoter region have H3R2me2s modification.Conclusion:PRMT5 was dynamically expressed during cerebrovascular development.H4R3 and H3R2 were substrate of Prmt5 in brain endothelial cells.The Bmp4 promoter region histone has H3R2me2s modification.And expression of Bmp4 was inhibited by Prmt5.

关 键 词：蛋白精氨酸甲基转移酶5(PRMT5) 血管发育 H3R2me2s 

分 类 号：Q25[生物学—细胞生物学] Q78