本文關(guān)鍵詞：PC-12神經(jīng)元作為海分枝桿菌感染宿主的實(shí)驗(yàn)研究　出處：《中國(guó)病原生物學(xué)雜志》2015年10期 　論文類型：期刊論文

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【摘要】：目的研究海分枝桿菌感染PC-12神經(jīng)元細(xì)胞的感染潛能及其誘導(dǎo)促炎性細(xì)胞因子表達(dá)的免疫特性。方法用海分枝桿菌感染神經(jīng)營(yíng)養(yǎng)因子(nerve growth factor,NGF)誘導(dǎo)分化的PC-12神經(jīng)元,采用激光共聚焦顯微鏡觀察細(xì)菌在PC-12神經(jīng)元細(xì)胞內(nèi)的定位分布,并計(jì)數(shù)細(xì)菌的菌落形成單位(CFU)以觀察其在PC-12神經(jīng)元細(xì)胞內(nèi)的增殖情況;采用實(shí)時(shí)熒光定量PCR(qRT-PCR)、酶聯(lián)免疫吸附試驗(yàn)(ELISA)及免疫印跡法(Western blot)檢測(cè)腫瘤壞死因子(tumor necrosis factor,TNF)-α、白介素(Interleukin)-1β、誘導(dǎo)型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和環(huán)氧合酶(cyclooxygenase-2,COX-2)mRNA及蛋白的表達(dá)情況。結(jié)果海分枝桿菌感染PC-12細(xì)胞后,激光共聚焦顯微鏡觀察細(xì)菌侵入PC-12神經(jīng)元胞漿內(nèi)并進(jìn)行增殖。當(dāng)細(xì)菌以MOI=20感染神經(jīng)元細(xì)胞時(shí),感染后24h與感染后6h相比,細(xì)菌在細(xì)胞內(nèi)增殖了約25倍。同時(shí)海分枝桿菌感染后6h即可誘導(dǎo)促炎性分子TNF-α、IL-1β、iNOS和COX-2的mRNA表達(dá)明顯增加,感染24h后TNF-α、IL-1β、iNOS和COX-2的蛋白表達(dá)也明顯增加。結(jié)論 PC-12能夠作為海分枝桿菌感染的宿主細(xì)胞,并引起促炎相關(guān)因子的分泌,從而促進(jìn)炎癥反應(yīng)。
[Abstract]:Objective to study the infection potential of PC-12 neurons infected with Mycobacterium sea and its immunological characteristics of inducing inflammatory cytokines expression. Nerve growth factor. PC-12 neurons were induced by NGF, and the localization and distribution of bacteria in PC-12 neurons were observed by confocal laser microscopy. The colony forming unit (CFU) of bacteria was counted to observe its proliferation in PC-12 neurons. The real-time fluorescence quantitative PCR qRT-PCR was used. Elisa and Western blot were used to detect tumor necrosis factor necrosis factor. TNF- 偽, Interleukin-1 尾, inducible nitric oxide synthase. Results the expression of iNOS) and cyclooxygenase-2 COX-2 mRNA and protein in PC-12 cells was observed. Laser confocal microscopy was used to observe the invasion of bacteria into the cytoplasm of PC-12 neurons and their proliferation. When the cells were infected with MOI=20 24 h after infection was compared with 6 h after infection. The bacteria proliferated about 25 times in the cells, and the mRNA expression of TNF- 偽, IL-1 尾, iNOS and COX-2 increased significantly 6 hours after infection. After 24 hours of infection, the expression of TNF- 偽 COX-2 and IL-1 尾 was also significantly increased. Conclusion PC-12 can be used as host cell of Mycobacterium kainiensis infection. And causes the secretion of pro-inflammatory related factors, thus promoting inflammatory response.
【作者單位】： 南通大學(xué)醫(yī)學(xué)院病原生物學(xué)系;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(No.81201252) 江蘇高校優(yōu)勢(shì)學(xué)科建設(shè)工程資助項(xiàng)目(No.PAPD) 江蘇省自然科學(xué)研究項(xiàng)目(14KJD180002,13KJB310013) 南通市科技計(jì)劃項(xiàng)目(No.HS2012059,BK2014033)
【分類號(hào)】：R378.911
【正文快照】： *?**?中樞神經(jīng)系統(tǒng)結(jié)核(central nervous system-tu-berculosis,CNS-TB)是由結(jié)核分枝桿菌(Mycobacte-rium tuberculosis,Mtb)穿越血腦屏障并入侵腦組織后引起的腦組織炎癥。該病主要多見于原發(fā)肺結(jié)核,其中在20%的肺結(jié)核患者中結(jié)核桿菌經(jīng)血行播散后發(fā)展成肺外結(jié)核,而肺外結(jié)核的

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