本文選題：綿羊肺炎支原體　切入點(diǎn)：莢膜多糖　出處：《寧夏大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：綿羊肺炎支原體(Mycoplasma ovipneuooniae,MO)是從綿羊或山羊支氣管中分離得到的一種最常見的病原菌,主要引起羊支原體肺炎�；疾⊙蚺R床上以漸進(jìn)性消瘦、高熱、氣喘、持續(xù)性咳嗽、肺和胸膜發(fā)生漿液性和纖維素性炎癥為特征。而與其他支原體相比,目前對(duì)于綿羊肺炎支原體致病機(jī)制的研究還非常有限,但普遍認(rèn)為準(zhǔn)確的黏附作用應(yīng)當(dāng)是決定其是否能夠成功感染的關(guān)鍵。綿羊肺炎支原體無(wú)細(xì)胞壁結(jié)構(gòu),感染宿主機(jī)體時(shí)主要通過膜外的莢膜樣物質(zhì)介導(dǎo)菌體與宿主細(xì)胞間的相互作用。已有報(bào)道證實(shí)莢膜多糖(capsularpolysaccharides,CPS)是位于綿羊肺炎支原體菌體最外層的一種重要毒力因子,其在菌體抗干燥、粘附、侵入、傳播以及抵抗宿主細(xì)胞的固有免疫和特異性免疫防御機(jī)制等方面的作用正被逐漸認(rèn)識(shí)。本研究在成功提取并純化了綿羊肺炎支原體模式株Y98莢膜多糖(CPS)的基礎(chǔ)上,制備了CPS的多克隆抗體,并利用綿羊支氣管原代上皮細(xì)胞氣液相(air-liquid interface,ALI)培養(yǎng)模型,對(duì)CPS誘發(fā)免疫學(xué)炎性反應(yīng)與氧化損傷過程中的分子機(jī)制進(jìn)行了探究。研究首先發(fā)現(xiàn),綿羊肺炎支原體CPS刺激ALI支氣管原代上皮細(xì)胞后誘導(dǎo)了 TLRs級(jí)聯(lián)信號(hào)通路介導(dǎo)的免疫學(xué)炎性反應(yīng)的發(fā)生。TLRs 信號(hào)中 MyD88、IRAKs、TRAF6、p-NF-κB、p-AP-1、IRF5 與 TRAF3、TBK1、IRF3等關(guān)鍵通路蛋白與核轉(zhuǎn)錄因子的表達(dá)量在CPS刺激后均呈現(xiàn)出明顯的上調(diào)趨勢(shì),表明TLRs級(jí)聯(lián)的MyD88依賴性與MyD88非依賴性信號(hào)轉(zhuǎn)導(dǎo)通路在支氣管上皮細(xì)胞響應(yīng)CPS刺激的過程中均扮演重要角色。研究同時(shí)發(fā)現(xiàn)CPS抗體的預(yù)處理可以顯著下調(diào)由綿羊肺炎支原體感染所誘導(dǎo)的炎性相關(guān)因子(包括促炎性細(xì)胞因子IL1β、IL6、IL8和TNFα等,抑炎性細(xì)胞因子IL10和TGFβ等)的高表達(dá),提示CPS是綿羊肺炎支原體的一個(gè)重要毒力因子,其致病機(jī)制可能是通過誘導(dǎo)支氣管周圍大量炎癥相關(guān)因子的釋放,從而實(shí)現(xiàn)對(duì)宿主免疫學(xué)炎癥反應(yīng)的調(diào)控。此外,分子機(jī)制研究表明綿羊肺炎支原體CPS可以通過線粒體介導(dǎo)的內(nèi)源性途徑與死亡受體介導(dǎo)的外源性途徑同時(shí)誘導(dǎo)支氣管上皮細(xì)胞的凋亡,JNK/P38MAPK信號(hào)通路在此過程中起重要的促凋亡作用。值得注意的是,CPS誘發(fā)宿主細(xì)胞凋亡的過程伴隨ROS的高表達(dá),ROS清除劑NAC的預(yù)處理可以明顯抑制這一現(xiàn)象的發(fā)生,因此提示ROS的過量產(chǎn)生可能是誘導(dǎo)綿羊支氣管原代上皮細(xì)胞凋亡的直接因素之一。以上的研究結(jié)果揭示,TLRs信號(hào)介導(dǎo)的炎性反應(yīng)與ROS介導(dǎo)的細(xì)胞凋亡在支氣管上皮細(xì)胞應(yīng)答綿羊肺炎支原體莢膜多糖刺激的免疫調(diào)控機(jī)制中發(fā)揮了關(guān)鍵性的作用。這些發(fā)現(xiàn)為進(jìn)一步研究綿羊肺炎支原體的主要毒力因子及其致病機(jī)理、闡明宿主機(jī)體的免疫應(yīng)答調(diào)控機(jī)制提供了新的思路。
[Abstract]:Mycoplasma pneumoniae (Mycoplasma ovipneuooniae MO) is one of the most common pathogens isolated from sheep or goats in bronchial, mainly caused by mycoplasma pneumonia. The prevalence of sheep sheep clinically with progressive weight loss, fever, persistent cough, asthma, lung and pleura had serous and fibrinous inflammation is characterized. Compared with other current research on Mycoplasma pneumoniae, Mycoplasma pneumoniae pathogenesis is still very limited, but it is generally believed that adhesion should be accurate to determine whether the key to success. No infection of Mycoplasma pneumoniae infection in the host cell wall structure, when the capsule like substances mainly through the membrane dielectric guide interaction with host cell between the cells. It has been reported that the capsular polysaccharide (capsularpolysaccharides, CPS) is an important virulence factor in Mycoplasma pneumoniae bacteria in the outer. Bacterial resistance to dry, adhesion, invasion, spread and resistance of host cell innate and specific immune defence mechanism function is being increasingly recognized. This study successfully extracted and Mycoplasma pneumoniae strain Y98 purified capsular polysaccharide (CPS) on the basis of the polyclonal antibody against CPS was prepared. And the use of primary bronchial epithelial cells (air-liquid, interface, ALI phase) culture model and molecular mechanism of CPS induced immunological inflammatory reaction and oxidative damage in the process were studied. Firstly, MyD88,.TLRs signal in mycoplasma pneumonia of sheep CPS immunological inflammatory stimuli ALI primary bronchial epithelial cells after the induction of TLRs cascade signaling mediated by IRAKs, TRAF6, p-NF- and IRF5 K B, p-AP-1, TRAF3, TBK1, IRF3 protein expression in the key pathway of nuclear transcription factor were found after CPS stimulation The upward trend is obvious, that plays an important role in TLRs cascade MyD88 dependent and MyD88 dependent signal transduction pathway in bronchial epithelial cells in response to CPS stimulation in the process. The study also found that CPS antibody pretreatment can significantly down regulate inflammatory factors induced by Mycoplasma pneumoniae infection (including pro-inflammatory cytokine IL1 beta, IL6, IL8 and TNF etc, suppression of inflammatory cytokines IL10 and TGF P) high expression, suggesting that CPS is an important virulence factor of Mycoplasma pneumoniae and its pathogenic mechanism may be through the relevant factor induced bronchial inflammation around massive release, so as to realize the control of the host the immunological inflammatory reaction. In addition, the molecular mechanism study indicated that exogenous endogenous pathway pathway Mycoplasma pneumoniae CPS can be mediated by mitochondria and death receptor mediated and induced by gas The apoptosis of tubular epithelial cells, JNK/P38MAPK signaling pathway in this process apoptosis is important. It is worth noting that the process of CPS induced apoptosis of host cells with high expression of ROS, ROS scavenger NAC pretreatment can significantly inhibit the occurrence of this phenomenon, therefore excessive production of ROS may be one of the direct factors of the original generation of sheep bronchial epithelial cell apoptosis. These results revealed that the apoptosis of inflammatory reaction and ROS mediated TLRs signaling mediated by play a key role in the regulation of immune response mechanism of bronchial epithelial cells of Mycoplasma pneumoniae capsular polysaccharide stimulation. These findings as the main virulence factors and its pathogenic mechanism further Mycoplasma ovipneumoniae, provides a new way to regulate the immune response mechanism of the host organism.

【學(xué)位授予單位】：寧夏大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S852.62

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王琪;汪洋;李媛;劉素麗;邵家日;陳瑩;高麗萍;周長(zhǎng)平;李春艷;辛九慶;;絲狀支原體絲狀亞種的脂質(zhì)相關(guān)蛋白通過MAPK信號(hào)通路誘導(dǎo)EBL細(xì)胞分泌IL-1β的研究[J];中國(guó)預(yù)防獸醫(yī)學(xué)報(bào);2016年02期

2 張會(huì)敏;馬宏;;肺炎支原體致病機(jī)制的研究進(jìn)展[J];中國(guó)現(xiàn)代醫(yī)生;2015年19期

3 鄧杰;張珂;袁濤;李春陽(yáng);;動(dòng)態(tài)顯色法檢測(cè)白喉毒素突變體CRM197的細(xì)菌內(nèi)毒素含量[J];中國(guó)生物制品學(xué)雜志;2013年12期

4 方積年;丁侃;;多糖的研究開發(fā)中值得注意的一些問題[J];食品與藥品;2007年12期

5 遲秀玲;嚴(yán)奉祥;;支原體引起細(xì)胞凋亡的機(jī)制[J];國(guó)際病理科學(xué)與臨床雜志;2006年06期

6 董碧麟,湯紀(jì)路,白瑞珍,宗潤(rùn)芝,劉先洲;肺炎支原體膜脂蛋白通過TLR2介導(dǎo)mICAM-1表達(dá)的上調(diào)[J];中華微生物學(xué)和免疫學(xué)雜志;2005年02期

7 韓張,胡萍,倪道鳳;短暫激活c-jun N-terminal激酶抑制膽紅素誘導(dǎo)的SH-SY5Y細(xì)胞凋亡[J];基礎(chǔ)醫(yī)學(xué)與臨床;2002年05期

8 蔣玉紅,張忠國(guó),王成;肺炎支原體肺炎患者外周血淋巴細(xì)胞凋亡過程及臨床意義[J];中國(guó)當(dāng)代兒科雜志;2001年05期

9 張啟敬;徐偉;李繼庚;王桂敏;丁慶猷;;豬肺炎霉形體兔化弱毒株的超微結(jié)構(gòu)及致病性研究——1.兔化弱毒株的超微結(jié)構(gòu)[J];電子顯微學(xué)報(bào);1989年04期

10 胡景韶,蔣學(xué)良,胡誠(chéng)隆,張道永;一種由支原體感染的綿羊增生性間質(zhì)性肺炎的研究[J];中國(guó)獸醫(yī)雜志;1982年05期

相關(guān)碩士學(xué)位論文 前6條

1 牛健;重組CARDs TX蛋白的克隆、表達(dá)與純化及多克隆抗體制備[D];天津醫(yī)科大學(xué);2014年

2 李雪靜;肺炎支原體外毒素CARDS TX的制備及特性研究[D];浙江大學(xué);2014年

3 倪博;豬肺炎支原體LAMPs誘導(dǎo)caspase3激活介導(dǎo)豬肺泡上皮細(xì)胞凋亡[D];安徽農(nóng)業(yè)大學(xué);2013年

4 周輝;沙眼衣原體pORF5質(zhì)粒蛋白經(jīng)TLR2激活MAPK信號(hào)通路誘導(dǎo)THP-1細(xì)胞產(chǎn)生前炎癥細(xì)胞因子[D];南華大學(xué);2012年

5 嚴(yán)勝平;CP5型金黃色葡萄球菌莢膜多糖的提取純化與鑒定[D];華中農(nóng)業(yè)大學(xué);2008年

6 陳宏偉;綿羊肺炎支原體的分離、鑒定及部分生物學(xué)特性的研究[D];石河子大學(xué);2005年

，

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