锰中毒小鼠黑质中m6A甲基化及其相关因子的表达研究  

Study on the expression of m6A methylation and its related factors in substantia nigra of mice with manganese poisoning

作　　者：廖嘉家 罗曼[1] 黄夏杰 黄婉 李滋虎 覃海兰 甘玉清 肖友生[1] 王进[1] Liao Jiajia;Luo Man;Huang Xiajie;Huang Wan;Li Zihu;Qin Hailan;Gan Yuqing;Xiao Yousheng;Wang Jin(Neurology Department,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院神经内科,南宁530021

出　　处：《广西医科大学学报》2022年第4期564-568,共5页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82060248,No.81801268);广西医科大学大学生创新创业计划项目(No.202010598156)。

摘　　要：目的:研究锰中毒小鼠黑质中m6A甲基化及其相关因子表达的变化。方法:将28只成年清洁级雄性昆明小鼠按照随机数字表法分成4组:对照组、MnCl_(2)低剂量组、中剂量组、高剂量组;对照组腹腔注射等量无菌生理盐水,MnCl_(2)低、中、高剂量组分别腹腔注射10 mg/kg、20 mg/kg、30 mg/kg的MnCl_(2)溶液,1次/d,连续4周,每天观察动物一般情况并记录体重变化。各组于注射结束后3 d取黑质组织,免疫荧光法观察黑质多巴胺能神经元损伤情况,m6A甲基化试剂盒检测各组m6A甲基化修饰水平,实时荧光定量聚合酶链式反应(RT-qPCR法)实验检测m6A甲基化相关基因在对照组和MnCl_(2)高剂量组小鼠黑质中的mRNA表达水平。结果:与对照组相比,MnCl_(2)高剂量组小鼠体重降低,黑质酪氨酸羟化酶(TH)阳性细胞数减少,甲基化水平、m6A促甲基化基因METTL3、去甲基化基因ALKBH5及FTO mRNA表达水平升高(均P<0.05)。结论:m6A甲基化水平及甲基化相关因子METTL3、ALKBH5和FTO在锰中毒小鼠黑质中高表达,提示m6A甲基化可能在锰中毒中发挥作用。Objective:To study the expressions of m6A methylation and its related factors in substantia nigra of mice with manganese poisoning.Methods:A total of 28 adult clean male Kunming mice were randomly divided into control group,MnCl_(2) low dose group,MnCl_(2) middle dose group and MnCl_(2) high dose group.The control group was intraperitoneally injected with the same amount of sterile saline,and the MnCl_(2) low,middle and high dose groups were intraperitoneally injected with 10 mg/kg,20 mg/kg and 30 mg/kg of MnCl_(2) solution respectively,once a day for 4 weeks,and the general condition of the mice was observed and the body weight was recorded.Three days after injection,the nigra tissue of each group was taken,and the damage of dopaminergic neurons in substantia nigra was observed by immunofluorescence.The level of m6A methylation modification was detected by m6A methylation kit.The mRNA expressions of m6A methylation-related genes in the substantia nigra of control group and MnCl_(2) high dose group were detected by RT-qPCR.Results:Compared with the control group,the body weight and the number of tyrosine hydroxylase(TH)positive cells decreased,and the methylation levels and the expressions of m6A methylation gene METTL3,demethylation gene ALKBH5 and FTO mRNA increased in the MnCl_(2) high dose group(P<0.05).Conclusion:The levels of m6A methylation and methylation-related factors METTL3,ALKBH5 and FTO are highly expressed in the substantia nigra of mice with manganese poisoning,suggesting that m6A methylation may play a role in manganese poisoning.

关 键 词：锰中毒 m6A甲基化 黑质 基因表达 

分 类 号：R742.5[医药卫生—神经病学与精神病学]