血管性痴呆模型大鼠大脑皮质N6-甲基腺苷甲基化修饰水平的变化  

The changes of N6 methyladenosine methylation in cortex of rat suffered from vascular dementia

作　　者：邓汝东[1] 周贤熙 周丽亭 周奕晴 翁安莉 张赛霞[1] 伍趣京[1] Deng Rudong;Zhou Xianxi;Zhou Liting;Zhou Yiqing;Weng Anli;Zhang Saixia;Wu Qujing(Center for Experimental Teaching,School of Basic Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006;Zhanjiang Traditional Chinese Medicine Hospital,Zhanjiang 524000,China)

机构地区：[1]广州中医药大学基础医学院实验教学中心,广州510006 [2]湛江市第一中医医院,湛江524000

出　　处：《神经解剖学杂志》2022年第5期572-576,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81673770)。

摘　　要：目的:研究N6-甲基腺苷甲基化(m6A)修饰参与血管性痴呆(VD)的机制。方法:36只SD雄性大鼠按照随机原则分为假手术组(sham)和血管性痴呆模型组(BCCAO),BCCAO大鼠结扎双颈总动脉,sham组大鼠不结扎双侧颈总动脉。免疫组织化学染色、real time RT-PCR和Western Blot检测大鼠大脑皮质YTH结构域蛋白1(YTHDF1)、YTH结构域蛋白2(YTHDF2)、YTH结构域蛋白3(YTHDF3)、去甲基化酶ALKB同源蛋白3(ALKBH3)、去甲基化酶ALKB5同源蛋白5(ALKBH5)和甲基转移酶样蛋白3(METTL3)的表达变化。结果:与sham组相比,免疫组织化学染色结果显示:BCCAO大鼠大脑皮质中YTHDF1、YTHDF2、YTHDF3、ALKBH3和ALKBH5阳性表达增加,METTL3阳性表达减少;RT-qPCR结果显示:BCCAO大鼠大脑皮质中YTHDF1、YTHDF2、YTHDF3、ALKBH3和ALKBH5 mRNA表达上升,METTL3 mRNA表达下降;Western Blot结果显示:BCCAO大鼠大脑皮质中YTHDF1、YTHDF2、YTHDF3、ALKBH3和ALKBH5蛋白质表达增加,METTL3蛋白质表达减少。结论:m6A甲基化修饰系统参与VD大鼠大脑皮质中的病理过程。Objective:To explore the mechanism of N6 methyladenosine methylation(m6 A)modification in vascular dementia.Methods:Thirty six adult male SD rats were randomly divided into sham group(sham),and vascular dementia model group(BCCAO).The BCCAO rats were obtained by bilateral carotid artery ligation.The expression of YT521 B homology domain family 1(YTHDF1),YT521 B homology domain family 2(YTHDF2),YT521 B homology domain family 3(YTHDF3),demethylases AlkB homologue 3(ALKBH3),demethylases AlkB homologue 5(ALKBH5)and methyltransferase like 3(METTL3)in cerebral cortex were detected by immunohistochemistry staining,real time RT-PCR,and Western Blot.Results:Immunohistochemical staining showed that the number of YTHDF1,YTHDF2,YTHDF3,ALKBH3,and ALKBH5 positive cells increased in BCCAO rats,the number of METTL3 positive cells decreased.The mRNA expression of YTHDF1,YTHDF2,YTHDF3,ALKBH3,and ALKBH5 in cerebral cortex of BCCAO rats was up-regulated.The protein expression of YTHDF1,YTHDF2,YTHDF3,ALKBH3,and ALKBH5 in cerebral cortex of BCCAO rats was increased.Meanwhile,the mRNA expression and the protein level of METTL3 were down-regulated on the contrary.Conclusion:The level of m6 A methylation was decreased in BCCAO rats.

关 键 词：N6-甲基腺苷甲基化 血管性痴呆 RNA甲基化 颈总动脉结扎 大鼠 

分 类 号：R749.13[医药卫生—神经病学与精神病学]