叶酸对SOD1-G93A小鼠的促炎作用研究  

Proinflammatory effect of folic acid on SOD1-G93A mice

作　　者：祝岩波 严陶陶 张双 周巍 Zhu Yanbo;Yan Taotao;Zhang Shuang;Zhou Wei(Hebei Food Inspection and Research Institute,Shijiazhuang 050227,China)

机构地区：[1]河北省食品检验研究院,石家庄050227

出　　处：《脑与神经疾病杂志》2024年第11期667-674,共8页Journal of Brain and Nervous Diseases

基　　金：河北省市场监管局科技计划项目(2022ZC14)。

摘　　要：目的探讨叶酸对肌萎缩侧索硬化症(ALS)的可能作用机制,为叶酸在临床上的防治提供可靠的实验数据。方法将转基因SOD1-G93A突变型小鼠12对随机分为对照组和叶酸组,同窝分开,雌雄各半,每组12只。从60 d开始,对照组正常进水进食,叶酸组正常进食,水添加4mg·kg^(-1)叶酸,每周称体质量2次。小鼠终末期脱颈处死后留取大脑、小脑、脊髓、心、肝、脾、肺、肾、坐骨神经、肌肉、小肠和结肠,提取RNA,PCR array试剂盒检测SOD1-G93A小鼠脊髓RNA表达水平,RT-qPCR检测SOD1-G93A小鼠大脑、小脑、脊髓、心、肝、脾、肺、肾、坐骨神经、肌肉、小肠和结肠中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、分化簇68(CD68)、分化簇86(CD86)和单核细胞趋化蛋白-1(MCP1)的表达水平。结果SOD1-G93A小鼠的叶酸组生存期为(128±8.1)d,对照组生存期为(136.5±9.7)d,叶酸组比对照组生存寿命平均减少8 d,差异有统计学意义(P<0.05);上调最为显著的基因有2个,分别为Epo和Bcl2a1a,涉及到Hypoxia和NF-κB信号通路。叶酸组在小脑和脊髓中TNF-α、IL-1β、CD68和CD 86的表达是对照组的两倍以上,叶酸组在小脑中MCP1的表达是对照组的两倍以上,差异有统计学意义(P<0.05)。叶酸组在心脏中TNF-α、IL-1β和MCP1的表达与对照组差异有统计学意义,在脾脏中IL-1β、CD68、CD86和MCP1的表达与对照组有显著性差异,其余无统计学意义。叶酸组在肌肉中TNF-α、CD68、CD86和MCP1的表达与对照组有显著性差异,叶酸组在结肠中TNF-α、IL-1β、CD68、CD86和MCP1的表达与对照组有显著性差异,其余差异无统计学意义。荧光染色结果表明SOD1-G93A小鼠脊髓中,Iba1和GFAP的表达叶酸组明显高于对照组。结论叶酸在SOD1-G93A小鼠模型中的干预可能激活了NF-κB信号通路,诱发了炎症反应,从而延长了病程。Objective To predict the possible mechanism of folic acid on amyotrophic lateral sclerosis(ALS)and to provide reliable experimental data for the prevention and treatment of folic acid in clinical practice.Methods Twelve pairs of transgenic SOD1-G93A mutant mice were randomly divided into control group and folic acid group,with 12 pairs in each group.Starting from 60 d,the control group ate normal water intake,the folic acid group ate normal food,adding 4mg·kg^(-1)folic acid to the water,weighing twice a week.RNA was extracted from the cerebrum,cerebellum,spinal cord,heart,liver,spleen,lung,kidney,sciatic nerve,muscle,small intestine and colon of mice after end-stage neck-removal and death.PCR array kit was used to detect the expression level of RNA in the spinal cord of SOD1-G93A mice.Tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and differentiation clusters in the cerebrum,cerebellum,spinal cord,heart,liver,spleen,lung,kidney,sciatic nerve,muscle,small intestine and colon of SOD1-G93A mice were detected by RT-qPCR expression levels of(CD68),differentiation cluster 86(CD86)and monocyte chemotactic protein-1(MCP1).Results The survival time of SOD1-G93A mice was(128±8.1)d in folic acid group and(136.5±9.7)d in control group,and the average survival time of folic acid group was reduced by8 days compared with control group,with statistically significant difference(P<0.05).Two of the most significantly upregulated genes,Epo and Bcl2ala,are involved in the Hypoxia and NF-κB signaling pathways.The expression of TNF-α,IL-1β,CD68 and CD86 in cerebellum and spinal cord of folic acid group was more than twice that of control group,and the expression of MCP1 in cerebellum of folic acid group was more than twice that of control group,with significant difference(P<0.05).The expressions of TNF-α,IL-1βand MCP1 in the heart of folic acid group were significantly different from those in the control group,and the expressions of IL-1β,CD68,CD86 and MCP1in the spleen were significantly different from those in the c

关 键 词：PCR array 叶酸 SOD1-G93A小鼠 炎症 

分 类 号：R744.8[医药卫生—神经病学与精神病学]