發(fā)布時(shí)間：2018-03-28 10:15

  本文選題：結(jié)核性腦膜炎　切入點(diǎn)：外泌體　出處：《鄭州大學(xué)》2017年碩士論文

【摘要】：背景與目的結(jié)核性腦膜炎(Tuberculoulous meningitis,TBM)是由結(jié)核分枝桿菌引起的中樞神經(jīng)系統(tǒng)感染性疾病,約占神經(jīng)系統(tǒng)結(jié)核病的70%左右。其目前診斷主要依靠腦脊液常規(guī)生化、細(xì)胞學(xué)檢查;然而在TBM發(fā)生的早期,腦脊液常規(guī)生化、細(xì)胞學(xué)變化多不典型,因此嚴(yán)重影響了TBM的早期診治,故探討TBM的潛在診斷標(biāo)志物具有重大的臨床意義。外泌體(exosomes)是一種由細(xì)胞主動(dòng)分泌的大小均一,直徑約30~100nm的膜性囊泡小體,具有脂質(zhì)雙分子層結(jié)構(gòu),廣泛存在于血液、尿液、唾液、腦脊液等體液中。其內(nèi)含有多種蛋白質(zhì)、mRNA、miRNA等,在不同細(xì)胞間進(jìn)行信息傳遞,發(fā)生多種生物功能。外泌體在中樞神經(jīng)系統(tǒng)中參與正常的生理?xiàng)l件以及介導(dǎo)病理狀態(tài),在中樞神經(jīng)系統(tǒng)的疾病的發(fā)生、發(fā)展及損傷修復(fù)中起著重要作用。微小RNA(mi RNA)是一類(lèi)內(nèi)源性非編碼的小分子單鏈RNA,其主要功能是從轉(zhuǎn)錄后水平調(diào)節(jié)基因的表達(dá),廣泛參與多種生物的生命過(guò)程。Let-7家族是miRNA重要的組成部分之一,其包含13個(gè)成員,其中l(wèi)et-7d在大腦中的表達(dá)水平非常高,在多種神經(jīng)系統(tǒng)疾病中起著重要作用,可能成為某些疾病的潛在生物學(xué)標(biāo)志物。本研究通過(guò)分離臨床上常見(jiàn)的病毒性、化膿性、結(jié)核性腦膜炎患者以及正常對(duì)照者腦脊液中的外泌體,來(lái)探討let-7d在四種腦脊液外泌體中的表達(dá)水平,以尋求TBM新的潛在診斷標(biāo)志物。材料與方法隨機(jī)收集我院15例結(jié)核性腦膜炎患者(結(jié)腦組,TBM組)、20例病毒性腦膜炎患者(病腦組,VM組)、10例化膿性腦膜炎患者(化腦組,PM組)以及20例正常腦脊液患者(對(duì)照組)的腦脊液。采取ExoQuick Precipitation提取法分離并純化外泌體。通過(guò)透射電鏡觀察其形態(tài),采用實(shí)時(shí)定量PCR法檢測(cè)其let-7d表達(dá)水平。結(jié)果1.三種腦炎患者及正常人腦脊液中均發(fā)現(xiàn)外泌體,電鏡下呈圓形、橢圓形或茶托狀外形,直徑約30~100nm,實(shí)時(shí)定量PCR法檢測(cè)發(fā)現(xiàn)其內(nèi)含有不同程度表達(dá)水平的let-7d。2.結(jié)核性腦膜炎患者腦脊液外泌體中l(wèi)et-7d的表達(dá)水平顯著低于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。3.let-7d表達(dá)水平在結(jié)腦組、化腦組與對(duì)照組相比均明顯下調(diào),差異具有統(tǒng)計(jì)學(xué)意義(P0.05);病腦組與對(duì)照組相比也下調(diào),但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。4.let-7d表達(dá)水平在化腦組與結(jié)腦組相比明顯下調(diào),差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。5.let-7d表達(dá)水平在結(jié)腦組與病腦組相比明顯下調(diào),差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論中樞神經(jīng)系統(tǒng)感染性疾病的腦脊液中可分離出外泌體,通過(guò)比較三種腦炎以及正常對(duì)照者外泌體中l(wèi)et-7d的表達(dá)水平,發(fā)現(xiàn)在TBM患者中l(wèi)et-7d的表達(dá)水平顯著下調(diào),為其診斷提供潛在的生物學(xué)標(biāo)志物。
[Abstract]:Background & objective: tuberculous meningitis (TBMM) is an infectious disease of the central nervous system caused by Mycobacterium tuberculosis, which accounts for about 70% of the tuberculosis of the nervous system. The diagnosis of TBMM mainly depends on routine biochemical and cytological examination of cerebrospinal fluid (CSF). However, in the early stage of TBM, the routine biochemical and cytological changes in cerebrospinal fluid (CSF) were atypical, which seriously affected the early diagnosis and treatment of TBM. Therefore, it is of great clinical significance to explore the potential diagnostic markers of TBM. Exocrine exosomes are a kind of membranous vesicles with the diameter of about 30~100nm, which is homogenous in size and active secreted by cells. It has a lipid bimolecular structure and widely exists in blood and urine. Saliva, cerebrospinal fluid and other body fluids contain a variety of proteins, such as mRNA-miRNA, which transmit information between different cells and produce a variety of biological functions. The exocrine body participates in normal physiological conditions and mediates the pathological state in the central nervous system. MicroRNAs play an important role in the development, development and repair of central nervous system diseases. MicroRNAs are a class of endogenous non-coding single-stranded RNAs, whose main function is to regulate the expression of genes at post-transcriptional level. The Let-7 family is one of the most important components of miRNA, which consists of 13 members, in which let-7d is highly expressed in the brain and plays an important role in a variety of neurological diseases. This study was conducted by isolating secretions from cerebrospinal fluid in patients with clinically common viral, suppurative, tuberculous meningitis and normal controls. To study the expression level of let-7d in four kinds of cerebrospinal fluid exocrine, Materials and methods Fifteen patients with tuberculous meningitis (tuberculous meningitis) were randomly collected from 15 patients with tuberculous meningitis (n = 20) with viral meningitis (n = 10) with suppurative meningitis (n = 10). Cerebrospinal fluid (CSF) of 20 normal cerebrospinal fluid patients (control group) were isolated and purified by ExoQuick Precipitation extraction method. Results 1. Exocrine bodies were found in cerebrospinal fluid of three kinds of encephalitis patients and normal subjects. The expression of let-7d in cerebrospinal fluid exocrine in tuberculous meningitis patients was significantly lower than that in control group, and the difference was statistically significant (P 0.05) .3.let-7d expression level of let-7d.2.The expression level of let-7d in tuberculous meningitis group was significantly lower than that in control group, and the expression level of let-7d in tuberculous meningitis group was significantly lower than that in control group. Compared with the control group, the expression of P0.05T in the control group was significantly lower than that in the control group (P 0.05), and the expression level of P0.05U. 4.let-7 d in the diseased brain group was lower than that in the control group, but there was no significant difference between the control group and the control group (P 0.05), and the expression level was significantly decreased in the control group (P < 0.05). The expression level of P0.05, 5.let-7d was significantly down-regulated in the nodal brain group compared with the diseased brain group, and the difference was statistically significant (P 0.05). Conclusion the exocrine can be separated from the cerebrospinal fluid of the infectious diseases of the central nervous system. By comparing the expression of let-7d in the exocrine of three encephalitis and normal controls, it was found that the expression of let-7d was significantly down-regulated in patients with TBM, which provided a potential biomarker for the diagnosis of encephalitis.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R529.3

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相關(guān)期刊論文 前1條

1 LI Chuan;ZHANG Jia;ZU Yu-Jiao;NIE Shu-Fang;CAO Jun;WANG Qian;NIE Shao-Ping;DENG Ze-Yuan;XIE Ming-Yong;WANG Shu;;Biocompatible and biodegradable nanoparticles for enhancement of anti-cancer activities of phytochemicals[J];Chinese Journal of Natural Medicines;2015年09期

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本文編號(hào)：1675890