mRNA N6⁃甲基腺苷甲基化在帕金森病患者血液中的变化

mRNA N6methyladenosine changes in the blood of patients with Parkinson’s disease

目的探究帕金森病(PD)患者血液中mRNAN6⁃甲基腺苷(m6A)甲基化水平以及相关的甲基转移酶[甲基转移酶样蛋白14(METTL14)]和去甲基化酶[脂肪与肥胖相关基因蛋白(FTO)、AlkB同源蛋白5(ALKBH5)]的变化情况。方法连续性收集2020年1月至2021年6月在该院神经内科门诊及住院就诊的PD患者(PD组,50例)以及健康对照人群(对照组,50例)。通过酶联免疫吸附测定(ELISA)、蛋白质印迹(WB)、实时定量PCR(QPCR)、m6A甲基化定量等实验,检测两组血浆及外周单个核细胞(PBMC)的METTL14、FTO、ALKBH5及m6A水平。结果与对照组比较,血浆中PD组的METTL14、FTO、ALKBH5蛋白水平较低,差异有统计学意义(P<0.05)。与对照组比较,外周单个核细胞(PBMC)中PD组的METTL14、FTO、ALKBH5蛋白条带灰度值较低,差异有统计学意义(P<0.05);与对照组比较,PBMC的mRNA中PD组的METTL14、FTO、ALKBH5表达水平较低,差异有统计学意义(P<0.05)。与对照组比较,PBMC的mRNA中PD组的m6A水平较低,差异有统计学意义(P<0.05)。结论在PD患者血浆和PBMC中MET⁃TL14、FTO、ALKBH5和m6A甲基化水平均有不同程度的下降,表明mRNAm6A甲基化与PD的发生发展存在一定的联系。

Objective To explore the changes in the levels of mRNA N6methyladenosine(m6A)and the related methyltransferase(methyltransferaselike protein 14[METTL14])and demethylases(fat mass and obesityassociated protein[FTO]and AlkB homolog 5[ALKBH5])in the blood of patients with Parkinson’s disease(PD)compared with healthy controls.Methods We consecutively enrolled 50 patients with PD at the inpatient and outpatient departments of neurology of the Affiliated Hospital of Jinan Medical University from January 2020 to June 2021,with 50 healthy participants as the control.The PD group and control group were compared with regard to the levels of METTL14,FTO,ALKBH5,and m6A in plasma and peripheral blood mononucleated cells(PBMC)by enzymelinked immunosorbent assay,Western blotting,quantitative realtime PCR,and m6A methylation quantification assay.Results Compared with the control group,the PD group showed significantly lower levels of METTL14,FTO,and ALKBH5 proteins in plasma and significantly lower gray values of METTL14,FTO,and ALKBH5 protein bands in PBMC(all P<0.05).The expression levels of METTL14,FTO,and ALKBH5 in PBMC mRNA were significantly lower in the PD group than in the control group(all P<0.05).The PD group had a significantly lower m6A level in PBMC mRNA than the control group(P<0.05).Conclusions Patients with PD have varying degrees of declines in METTL14,FTO,ALKBH5,and m6A levels in plasma and PBMC,suggesting a certain link between m6A and the development and progression of PD.