首页> 外文期刊>Microbial Pathogenesis >Enteroaggregative Escherichia coli induced increase in intracellular calcium concentration modulates cytoskeletal F-actin rearrangement and bacterial entry in INT-407 cells.
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Enteroaggregative Escherichia coli induced increase in intracellular calcium concentration modulates cytoskeletal F-actin rearrangement and bacterial entry in INT-407 cells.

机译:肠聚合性大肠杆菌诱导的细胞内钙浓度增加调节了INT-407细胞中细胞骨架F-肌动蛋白的重排和细菌进入。

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Background: Enteroaggregative Escherichia coli (EAEC) is an emerging enteric pathogen, associated with cases of acute and persistent diarrhoea worldwide. The pathogenesis of EAEC is yet to be understood. In intestinal epithelium, an increase in [Ca2+]i has been attributed due to the action of different enteric pathogens. EAEC was shown to increase [Ca2+]i in HEp-2 cells.The present study was undertaken to investigate the effect of EAEC induced increase in [Ca2+]i oncultured human intestinal epithelial cells. Methods: INT-407 cells were infected with EAEC (T8 strain) in the absence and presence of dantrolene (inhibitor of release of Ca2+ from intracellular stores)/verapamil (L-type Ca2+ channel blocker)/BAPTA-AM (Ca2+ chelator)/U73122 (PLC inhibitor)/Cytochalasin-D (inhibitor of actin polymerization). [Ca2+]i was estimated using Fura-2/AM. Cytoskeletal rearrangement was assessed by F-actin staining using TRITC-phalloidin. The invasiveness of EAEC-T8 to INT-407 cells was checked by electron microscopy and invasion assay. Results: A significant increase in [Ca2+]i was observed in EAEC-T8 infected INT-407 cells, which was reduced in presence of dantrolene/verapamil/U73122. EAEC-T8 could induce cytoskeletal F-actin polymerization in INT-407 cells and was found to be invasive in nature. The cytoskeletal rearrangement as well as invasion of EAEC-T8 was attenuated in presence of U73122/dantrolene/BAPTA-AM/verapamil/cytochalasin D. Conclusions: EAEC induced increase in [Ca2+]i seems to play a major role in host cytoskeletal F-actin rearrangements leading to invasion of the organism. General significance: Our study undoubtedly will lead to an improved understanding of EAEC-pathogenesis.
机译:背景:肠道聚集性大肠埃希氏菌(EAEC)是一种新兴的肠道病原体,与全球范围内的急性和持续性腹泻有关。 EAEC的发病机制还有待了解。在肠道上皮中,[Ca 2 + ] i 的增加归因于不同肠病原体的作用。 EAEC在HEp-2细胞中可增加[Ca 2 + ] i 。本研究旨在研究EAEC诱导的[Ca 2+ 2 + ] i 培养的人肠上皮细胞。方法:在不存在和存在丹特林(抑制细胞内Ca 2 + 释放的抑制剂)/维拉帕米(L型Ca )的情况下,用EAEC(T8株)感染INT-407细胞2 + 通道阻滞剂)/ BAPTA-AM(Ca 2 + 螯合剂)/ U73122(PLC抑制剂)/细胞松弛素-D(肌动蛋白聚合抑制剂)。使用Fura-2 / AM估算[Ca 2 + ] i 。使用TRITC-鬼笔环肽通过F-肌动蛋白染色评估细胞骨架重排。 EAEC-T8对INT-407细胞的侵袭性通过电子显微镜和侵袭试验进行了检查。结果:在被EAEC-T8感染的INT-407细胞中观察到[Ca 2 + ] i 的显着增加,但在丹特罗林/维拉帕米/ U73122的存在下降低了。 EAEC-T8可以诱导INT-407细胞中的细胞骨架F-肌动蛋白聚合,并被发现具有侵入性。在U73122 / dantrolene / BAPTA-AM / verapamil / cytochalasin D的存在下,细胞骨架重排以及对EAEC-T8的侵袭减弱。结论:EAEC诱导[Ca 2 + ] 的增加i 似乎在宿主细胞骨架F-肌动蛋白重排导致该生物入侵方面起着重要作用。一般意义:毫无疑问,我们的研究将使人们对EAEC发病机理有更深入的了解。

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