本文選題：空腸彎曲菌 + GBS�。� 參考：《揚州大學(xué)》2012年碩士論文

【摘要】：空腸彎曲菌Campylobacter jejuni(CJ),是人類腸胃炎的最主要的致病菌,臨床約90%的急性細(xì)菌腸炎均與該菌相關(guān)。在歐美等國,其致病率已超過沙門氏菌、志賀氏菌、O157大腸桿菌等。CJ除了引起腸炎之外它還是是吉蘭-巴雷綜合征Guillain-Barre syndrome (GBS)的主要誘因之一,有報道指出,在CJ感染之后人類GBS的發(fā)病率會提升100倍。GBS又稱急性炎癥性脫髓鞘性多發(fā)性神經(jīng)病(AIDP),是神經(jīng)系統(tǒng)由體液和細(xì)胞共同介導(dǎo)的單向性自身免疫性周圍神經(jīng)病,死亡率約5～10%。鑒于CJ極高的感染率以及后續(xù)的潛在威脅,已受到人們的密切關(guān)注。本研究利用巴馬小型豬構(gòu)建空腸彎曲菌誘導(dǎo)GBS的動物模型,挖掘GBS模式動物中與病發(fā)相關(guān)的基因。 1空腸彎曲菌致GBS動物模型的建立 選取致人GBS源空腸彎曲菌株,采用3×1012cfu/ml、3×1011cfu/ml、3×1010cfu/ml、3×109cfu/ml等濃度對4組巴馬小型豬灌胃攻毒,每頭豬攻毒10ml,同時設(shè)立對照組。從第二周開始定期采集組織樣及血液樣本,通過免疫學(xué)檢測及組織病理切片判定病變情況。免疫學(xué)檢測發(fā)現(xiàn),在16份攻毒組樣品中有12份血清中GM1-IgG抗體為檢測陽性,陽性檢出率為81.25%。但是,并沒有顯著的攻毒濃度梯度差異。病理組織切片H/E染色顯示,抗體陽性試驗豬的大腦出現(xiàn)了噬神經(jīng)元、小血管周圍間隙增寬、神經(jīng)中央染色溶解等現(xiàn)象；切片LFB染色顯示,在發(fā)病模型動物的小腦白質(zhì)和腰膨大出現(xiàn)了輕度脫髓鞘現(xiàn)象。結(jié)果表明空腸彎曲菌誘導(dǎo)GBS的濃度在3X109cfVml-3X1012cfu/ml均可,組織樣本及血液采集在攻毒后4～5周較理想。 2空腸彎曲菌致GBS病發(fā)時宿主體內(nèi)相關(guān)基因的研究 根據(jù)病理組織切片和免疫學(xué)檢測結(jié)果,選取典型發(fā)病樣本和對照組樣本的神經(jīng)組織。通過豬全基因組表達(dá)譜芯片檢測得到差異探針2156個(P0.05),其中上調(diào)差異探針896條,下調(diào)差異探針1260條,這些探針涉及到的基因共222個,其中上調(diào)基因114個,下調(diào)基因108個�；贕ene Ontology數(shù)據(jù)庫對222個差異基因進(jìn)行分析,得到顯著基因功能(GO)130個(P0.05),其中上調(diào)基因顯著性GO55個,下調(diào)基因顯著性GO75個；基于KEGG數(shù)據(jù)庫,分析所有差異基因參與的信號通路(Pathway),得到相關(guān)Pathway共127項,其中上調(diào)基因相關(guān)的顯著性Pathway30項,下調(diào)基因相關(guān)的顯著性Pathway37項；采用顯著性GO內(nèi)所含的基因與顯著性Pathway所含的基因,對二者取交集,使用交集內(nèi)的基因分別構(gòu)建病變組和對照組的共表達(dá)網(wǎng)絡(luò),最終得到21個關(guān)鍵基因。其中HPRT1、PKC和PPARGC-1等處于網(wǎng)絡(luò)圖的核心位置,PPARGC-1、SUS2DD、AMPKA2等11個關(guān)鍵基因在病發(fā)時都表現(xiàn)出顯著性被抑制的狀態(tài)。
[Abstract]:Campylobacter jejunii CJ is the main pathogen of human gastroenteritis, and 90% of acute bacterial enteritis is associated with it. In Europe, America and other countries, the pathogenic rate of Guillain-Barre syndrome is higher than that of Salmonella. Besides causing enteritis, E. coli. CJ of Shigella spp. O157 is also one of the main causes of Guillain-Barre syndrome. After CJ infection, the incidence of GBS in human will increase 100 times. GBS is also called acute inflammatory demyelinating polyneuropathy (AIDP). It is a unidirectional autoimmune peripheral neuropathy mediated by body fluid and cells in the nervous system. The death rate is about 510g. In view of the extremely high infection rate of CJ and its potential threat, people have paid close attention to it. In this study, the animal model of GBS induced by Campylobacter jejuni was constructed from Bama miniature pig. 1 the establishment of Campylobacter jejuni animal model induced by Campylobacter jejuni caused by human Campylobacter jejuni. 3 脳 1012cfur / ml 3 脳 1011cfuml 3 脳 1010cfuml / ml 3 脳 1010cfuml / ml 3 脳 10 10cfuml / ml 3 脳 109cfu/ml were administered to 4 groups of small pigs. Each pig was treated with 10 ml of virus, and the control group was set up at the same time. Tissue samples and blood samples were collected regularly from the second week, and pathological changes were determined by immunological examination and histopathological section. Immunological examination showed that the GM1-IgG antibody was positive in 12 of the 16 samples of the group, and the positive rate was 81.25%. However, there was no significant difference in concentration gradient. Histopathological sections showed that phagocytic neurons were present in the brain of pigs with positive antibodies, the spaces around small vessels were widened, and central nerve staining was dissolving, and LFB staining showed that, Mild demyelination of cerebellar white matter and lumbar swelling was observed in the model animals. The results showed that the concentration of GBS induced by Campylobacter jejuni was at 3X109cfVml-3X1012cfur / ml. Tissue samples and blood samples were collected at 4 to 5 weeks after attack. 2 the study of genes related to GBS caused by Campylobacter jejuni according to the results of histopathological sections and immunological examination. The nerve tissue of the typical disease and the control group were selected. A total of 2156 differentially expressed probes were detected by porcine genomic expression microarray, of which 896 were up-regulated and 1260 were down-regulated. These probes involved 222 genes, including 114 up-regulated genes and 108 down-regulated genes. Based on the analysis of 222 differentially expressed genes based on Gene Ontology database, 130 significant gene functions (P0.05) were obtained, of which 55 genes were up-regulated and 75 genes were down-regulated, which were based on KEGG database. A total of 127 related Pathway genes were obtained by analyzing the signal pathways involved in all the differentially expressed genes, among which 30 were up-regulated and 37 were down-regulated, and the genes contained in significant go and those contained in significant Pathway were used. The gene in the intersection was used to construct the coexpression network of the diseased group and the control group. Finally, 21 key genes were obtained. Among them, 11 key genes such as HPRT1PKC and PPARGC-1, such as PPARGC-1, SUS2DDD, AMPKA2 and so on, showed significant inhibition at the onset of the disease.
【學(xué)位授予單位】：揚州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R-332;R379.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 趙曉燕,薛平,劉瑞春,李春巖,李艷華,賈麗梅,王桂榮,楊程;空腸彎曲菌抗體對兔周圍神經(jīng)致病作用的研究[J];中風(fēng)與神經(jīng)疾病雜志;1999年06期

2 王愛德;廣西巴馬小型豬的選育研究[J];中國比較醫(yī)學(xué)雜志;2004年03期

3 張建中,李春巖,陳晶晶,劉瑞春,蔣秀高,田文強;格林-巴利綜合征相關(guān)空腸彎曲菌脂多糖抗原初步分析[J];中華微生物學(xué)和免疫學(xué)雜志;1997年02期

，

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