问号钩端螺旋体内化过程中细胞骨架变化及相关信号通路

发布时间：2018-01-25 19:29

本文关键词： 问号钩端螺旋体黏附内化细胞骨架 F-actin 磷脂酶C　出处：《浙江大学》2006年硕士论文　论文类型：学位论文

【摘要】：背景和目的钩端螺旋体(简称钩体)病是全世界流行的人兽共患病,我国也是钩体病流行的主要疫区。钩体通过皮肤黏膜,迅速进入血液,并在血液和组织内生长繁殖,造成机体损伤,引起一系列症状和体征。其临床综合症包括亚临床感染、伴或不伴有脑膜炎的无黄疸的自限性发热、潜在性致命的以出血、黄疸、肾衰竭为表现的Weil's综合症。钩体在感染动物肾脏组织内长期生存并不断随尿排出,提示钩体必然具有侵入宿主细胞的能力,但确切的致病机制至今未明。有文献报道,问号钩体可黏附并侵入细胞,进一步研究结果证实,强毒力和弱毒力问号钩体均能侵入J774A.1及Vero细胞,前者还可侵入细胞核,而无毒力的双曲钩体缺乏侵入细胞的能力。近年来报道,许多致病菌黏附宿主细胞后能引起Ca~(2+)升高,Ca~(2+)可以触发信号传导机制,诱导宿主细胞骨架重排,最终导致细菌内化入细胞。钩体内化细胞时能否触发信号传导机制,诱导宿主细胞骨架重排,尚需要研究。在以往工作基础上,本研究用微丝(F-actin)的特异性标记物,罗丹明标记的鬼笔环肽(Phalloidin-TRITC)染F-actin,用抗微管蛋白α亚单位的小鼠一抗(Anti-Bovine α-Tubulin,Mouse Monoclonal)和荧光素标记的羊抗小鼠二抗(FITC-conjugated AffniPure Goat Anti-Mouse IgG)染微管(microtubule),检测了不同毒力的钩体侵入宿主细胞时细胞骨架的改变情况,并用磷脂酶C(PLC)信号传导分子的特异性阻断剂U73122抑制磷脂酶C信号通路,观察其对细胞骨架改
[Abstract]:Background and objective Leptospira leptospirosis (Leptospirosis) is the most prevalent zoonosis in the world. Leptospirosis is also the main epidemic area of leptospirosis in China. Leptospirosis enters the blood rapidly through the skin mucosa. It also grows and multiplies in blood and tissue, causing damage to the body and causing a series of symptoms and signs. Its clinical syndrome includes subclinical infection, self-limited fever without jaundice with or without meningitis. Potentially fatal Weil's syndrome characterized by bleeding, jaundice, and renal failure. Leptospira survives in the kidneys of infected animals for a long time and is constantly excreted with urine. It is suggested that Leptospira must have the ability to invade host cells, but the exact pathogenetic mechanism is still unknown. It has been reported that Leptospira interrogans can adhere to and invade cells. Further studies have confirmed that both highly virulent and weakly virulent Leptospira can invade J774A.1 and Vero cells, and the former can also invade the nucleus. In recent years, it has been reported that the adhesion of many pathogenic bacteria to host cells can cause the increase of Ca~(2), which can trigger the signal transduction mechanism. Inducement of cytoskeleton rearrangement of host cells leads to bacterial internalization into cells. Whether Leptospira internalization can trigger signal transduction mechanism and induce cytoskeleton rearrangement of host cells needs to be studied. On the basis of previous work, F-actin was stained with Rhodamine labeled Phalloidin-TRITC, a specific marker of F-actinin. Anti-Bovine 伪 -Tubulin was used in mice with anti-tubulin 伪 subunit. Mouse Monoclonal) and fluorescein labeled sheep anti mouse second antibody. FITC-conjugated AffniPure Goat Anti-Mouse IgG (microtubule). The cytoskeleton changes of Leptospira with different virulence in host cells were detected, and the phospholipase C signaling pathway was inhibited by U73122, a specific inhibitor of phospholipase C (PLC) signal transduction molecule. Observation of cytoskeleton modification
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R377

【参考文献】