结核性脑膜炎患者脑脊液外泌体中let-7d表达的研究  

Study of the expression of let-7d from cerebrospinal fluid exosomes in patients with tuberculous meningitis

作　　者：尹慧敏[1] 贾永林[1,2] 李燕飞 段冉冉[1] 景黎君[1] 滕军放[1] 贾延劼[1] 

机构地区：[1]郑州大学第一附属医院神经内科,郑州450052 [2]河南开封市中心医院神经内科,开封475000

出　　处：《中国实用神经疾病杂志》2017年第6期9-12,共4页Chinese Journal of Practical Nervous Diseases

基　　金：国家自然科学基金(编号:81371385)

摘　　要：目的探讨结核性脑膜炎(tuberculoulous meningitis,TBM)患者脑脊液外泌体的分离及外泌体中let-7d的表达水平及其意义。方法收集我院15例结核性脑膜炎患者(结脑组,TBM组),20例病毒性脑膜炎患者(病脑组,VM组),10例化脓性脑膜炎患者(化脑组,PM组)及20例正常脑脊液患者(对照组)的脑脊液,采取ExoQuick Precipitation提取法分离并纯化外泌体,通过透射电镜观察其形态,采用实时荧光定量PCR法检测其let-7d表达水平。结果 (1)脑脊液中的外泌体,电镜下呈圆形或茶托状外形,直径40~100nm,实时定量PCR法检测发现其内含有不同程度表达水平的let-7d;(2)结核性脑膜炎患者脑脊液外泌体中let-7d的表达水平显著低于对照组,差异有统计学意义(P<0.05);(3)let-7d表达水平结脑组、化脑组与对照组相比均明显下调,差异有统计学意义(P<0.05);病脑组与对照组相比也下调,但差异无统计学意义(P>0.05);(4)let-7d表达水平在化脑组与结脑组相比明显下调,差异有统计学意义(P<0.05);(5)let-7d表达水平在结脑组与病脑组相比明显下调,差异有统计学意义(P<0.05)。结论中枢神经系统感染性疾病的脑脊液中可分离出外泌体,通过比较3种脑炎外泌体中let-7d的表达水平,发现在TBM患者中let-7d的表达水平显著下调,为其诊断提供潜在的生物学标志物。Objective To investigate the exosomes isolation from cerebrospinal fluid(CSF)in patients with tuberculous meningitis(TBM)and the expression of let-7din exosomes.Methods The cerebrospinal fluid from 15 cases with tuberculous meningitis(TBM group),20 cases with viral meningitis(VM group),10 cases with purulent meningitis patients(PM group)and 20 normal cases(control group)was collected.ExoQuick Precipitation was used to isolate and purify exosomes and transmission electron microscopy was applied to observe the morphology.Eventually,the expression of let-7dwas detected by real-time fluorescence quantitative PCR.Results(1)The exosomes in cerebrospinal fluid were round or cup-shaped,and the diameter was about 40~100nm.Real-time quantitative PCR found varying expression levels of let-7din exosomes.(2)The expression level of let-7din TBM group was significantly lower than that in control group(P<0.05).(3)The TBM and PM groups showed higher down-regulation of the expression level of let-7dcompared with the control group(P<0.05).(4)The expression level of let-7din the PM group was significantly down-regulated relative to the TBM group(P<0.05).(5)Significant down-regulation of expression level of let-7d was found in TBM group compared with VM group(P<0.05).Conclusion Exosomes can be isolated from cerebrospinal fluid in infectious diseases of the central nervous system.By comparing the expression level of let-7din three sorts of encephslitis,we find that the expression of let-7din TBM patients is significantly down-regulated,providing apotential biomarker for diagnosis.

关 键 词：结核性脑膜炎 外泌体 分离 let-7d 

分 类 号：R529.3[医药卫生—内科学]