VDR启动子DNA甲基化在EAE神经损伤中的作用研究  

Role of VDR Promoter DNA Methylation in Experimental Autoimmune Encephalomyelitis Nerve Injury

作　　者：朱俊羽 楚兰[1] 徐竹 高子丹 邵冰 丹珍卓玛 ZHU Jun-yu;CHU Lan;XU Zhu;GAO Zi-dan;SHAO Bing;Danzhenzhuoma(Guizhou Medical University,Guiyang 550004,China;Department of Neurology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学,贵阳550004 [2]贵州医科大学附属医院神经内科,贵阳550004

出　　处：《神经损伤与功能重建》2022年第9期497-501,508,共6页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金(No.81960231)。

摘　　要：目的:研究维生素D受体(VDR)启动子DNA甲基化对实验性自身免疫性脑脊髓炎(EAE)病程的影响。方法:C57BL/6雌性小鼠,分为对照组(n=12)和EAE组(n=48),EAE组通过注射MOG35-55建立EAE模型,对照组不予任何处理。EAE组按Weaver’s 15分神经功能评分法分为0分亚组(n=10)、高分亚组(n=10)、低分亚组(n=10)。荧光定量PCR(QPCR)和免疫印迹(WB)检测VDR在各组小鼠脊髓腰段的表达水平;亚硫酸钠测序技术(BSP)检测各组VDR启动子DNA甲基化水平。结果:在透射电镜下观察EAE 0分亚组、高分亚组、低分亚组小鼠髓鞘结构,较对照组都有不同程度的破坏:0分亚组、低分亚组髓鞘环形结构存在,高分亚组髓鞘环形结构消失。在脊髓组织中,EAE 0分亚组、高分亚组、低分亚组VDR蛋白表达都低于对照组(P_(0分亚组)=0.0046;P_(高分亚组)=0.0088;P_(低分亚组)<0.0001);低分亚组较0分亚组进一步下降(P=0.0032),同时低分亚组较高分亚组也呈下降趋势(P=0.0134)。EAE各组VDR mRNA表达也呈下降趋势,都低于对照组(P_(0分亚组)>0.05;P_(高分亚组)=0.0439;P_(低分亚组)=0.0349),与0分亚组比较,低分亚组减少明显(P=0.0142)。各组小鼠VDR启动子DNA甲基化率都较低,且无变化趋势,差异无统计学意义(P>0.05)。结论:VDR在EAE模型的不同神经功能评分亚组之间有表达差异,VDR启动子DNA的甲基化不是导致这些差异的主要原因。Objective: To investigate effect of vitamin D receptor (VDR) promoter DNA methylation on the course of experimental autoimmune encephalomyelitis (EAE). Methods: C57BL/6 female mice were selected to the control group (n=12) or EAE group (n=48). The EAE model was established in the EAE group by injection of MOG35-55. The control group received no treatment. Using the 15-point Weaver’s neurological function scoring method, the EAE group was divided into the 0 score (n=10), high score (n=10), and low score (n=10) subgroups. Quantitative real time PCR (qPCR) and Western blot (WB) were used to detect the expression level of VDR in the lumbar spinal cord of mice in each group. Bisulfite genomic sequencing (BSP) was used to detect the DNA methylation level of the VDR promoter in each group. Results: The myelin sheath structure of the 0 score, high score, and low score subgroup mice was observed under transmission electron microscope and showed different degrees of damage when compared to the control group. The myelin sheath ring structure of the 0 score and low score subgroups was intact, and that of the high score subgroup disappeared. In the spinal cord, the expression of VDR protein in the EAE 0 score, high score, and low score subgroups was lower than that in the control group (P_(0)=0.0046, P_(high)=0.0088, P_(low)<0.0001);in the low score subgroup was further lower than in the 0 score subgroup (P=0.0032), and in the low score subgroup was lower than in the high score subgroup (P= 0.0134). The expression of VDR mRNA in each EAE subgroup also showed a downward trend compared to that in the control group (P_(0)>0.05, P_(high)=0.0439, Plow=0.0349);compared with expression in the 0 score subgroup, that in the low score subgroup decreased significantly (P=0.0142). The DNA methylation level of the VDR promoter in each group was low, there was no change in trend, and the difference was not statistically significant (P>0.05). Conclusion: VDR was expressed differently in different neural function score groups of the EAE mo

关 键 词：维生素D受体 DNA甲基化 实验性自身免疫性脑脊髓炎 脊髓 

分 类 号：R741[医药卫生—神经病学与精神病学] R744.51[医药卫生—临床医学]