PC-12神经元作为海分枝杆菌感染宿主的实验研究

发布时间：2018-01-04 04:00

本文关键词：PC-12神经元作为海分枝杆菌感染宿主的实验研究　出处：《中国病原生物学杂志》2015年10期 　论文类型：期刊论文

【摘要】：目的研究海分枝杆菌感染PC-12神经元细胞的感染潜能及其诱导促炎性细胞因子表达的免疫特性。方法用海分枝杆菌感染神经营养因子(nerve growth factor,NGF)诱导分化的PC-12神经元,采用激光共聚焦显微镜观察细菌在PC-12神经元细胞内的定位分布,并计数细菌的菌落形成单位(CFU)以观察其在PC-12神经元细胞内的增殖情况;采用实时荧光定量PCR(qRT-PCR)、酶联免疫吸附试验(ELISA)及免疫印迹法(Western blot)检测肿瘤坏死因子(tumor necrosis factor,TNF)-α、白介素(Interleukin)-1β、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和环氧合酶(cyclooxygenase-2,COX-2)mRNA及蛋白的表达情况。结果海分枝杆菌感染PC-12细胞后,激光共聚焦显微镜观察细菌侵入PC-12神经元胞浆内并进行增殖。当细菌以MOI=20感染神经元细胞时,感染后24h与感染后6h相比,细菌在细胞内增殖了约25倍。同时海分枝杆菌感染后6h即可诱导促炎性分子TNF-α、IL-1β、iNOS和COX-2的mRNA表达明显增加,感染24h后TNF-α、IL-1β、iNOS和COX-2的蛋白表达也明显增加。结论 PC-12能够作为海分枝杆菌感染的宿主细胞,并引起促炎相关因子的分泌,从而促进炎症反应。
[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- 伪, 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- 伪, IL-1 尾, iNOS and COX-2 increased significantly 6 hours after infection. After 24 hours of infection, the expression of TNF- 伪 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.
【作者单位】：南通大学医学院病原生物学系;
【基金】：国家自然科学基金项目(No.81201252) 江苏高校优势学科建设工程资助项目(No.PAPD) 江苏省自然科学研究项目(14KJD180002,13KJB310013) 南通市科技计划项目(No.HS2012059,BK2014033)
【分类号】：R378.911
【正文快照】： *?**?中枢神经系统结核(central nervous system-tu-berculosis,CNS-TB)是由结核分枝杆菌(Mycobacte-rium tuberculosis,Mtb)穿越血脑屏障并入侵脑组织后引起的脑组织炎症。该病主要多见于原发肺结核,其中在20%的肺结核患者中结核杆菌经血行播散后发展成肺外结核,而肺外结核的

【参考文献】