Chen Zhixiong, Suntres Zach, Palmer Jeffrey, Guzman Jorge, Javed Asad, Xue Jianjing, Yu Jun-Ge, Cooke Helen, Awad Hamdy, Hassanain Hamdy H, Cardounel Arturo J, Christofi Fievos L
Department of Anesthesiology, The Ohio State University, Columbus, OH 43210, USA.
Int J Parasitol. 2007 Jun;37(7):743-61. doi: 10.1016/j.ijpara.2006.12.018. Epub 2007 Jan 14.
Trichinella spiralis infection causes hyperexcitability in enteric after-hyperpolarising (AH) sensory neurons that is mimicked by neural, immune or inflammatory mediators known to stimulate adenylyl cyclase (AC)/cyclic 3',5'-adenosine monophosphate (cAMP) signaling. The hypothesis was tested that ongoing modulation and sustained amplification in the AC/cAMP/phosphorylated cAMP related element binding protrein (pCREB) signaling pathway contributes to hyperexcitability and neuronal plasticity in gut sensory neurons after nematode infection. Electrophysiological, immunological, molecular biological or immunochemical studies were done in T. spiralis-infected guinea-pigs (8000 larvae or saline) after acute-inflammation (7 days) or 35 days p.i., after intestinal clearance. Acute-inflammation caused AH-cell hyperexcitability and elevated mucosal and neural tissue levels of myeloperoxidase, mast cell tryptase, prostaglandin E2, leukotrine B4, lipid peroxidation, nitric oxide and gelatinase; lower level inflammation persisted 35 days p.i. Acute exposure to blockers of AC, histamine, cyclooxygenase or leukotriene pathways suppressed AH-cell hyperexcitability in a reversible manner. Basal cAMP responses or those evoked by forskolin (FSK), Ro-20-1724, histamine or substance P in isolated myenteric ganglia were augmented after T. spiralis infection; up-regulation also occurred in AC expression and AC-immunoreactivity in calbindin (AH) neurons. The cAMP-dependent slow excitatory synaptic transmission-like responses to histamine (mast cell mediator) or substance P (neurotransmitter) acting via G-protein coupled receptors (GPCR) in AH neurons were augmented by up to 2.5-fold after T. spiralis infection. FSK, histamine, substance P or T. spiralis acute infection caused a 5- to 30-fold increase in cAMP-dependent nuclear CREB phosphorylation in isolated ganglia or calbindin (AH) neurons. AC and CREB phosphorylation remained elevated 35 days p.i.. Ongoing immune activation, AC up-regulation, enhanced phosphodiesterase IV activity and facilitation of the GPCR-AC/cAMP/pCREB signaling pathway contributes to T. spiralis-induced neuronal plasticity and AH-cell hyperexcitability. This may be relevant in gut nematode infections and inflammatory bowel diseases, and is a potential therapeutic target.
旋毛虫感染会导致肠道后超极化(AH)感觉神经元出现兴奋性过高,已知能刺激腺苷酸环化酶(AC)/环磷酸腺苷(cAMP)信号传导的神经、免疫或炎症介质可模拟这种情况。本研究检验了以下假设:AC/cAMP/磷酸化环磷酸腺苷反应元件结合蛋白(pCREB)信号通路中持续的调节和持续的放大作用,有助于线虫感染后肠道感觉神经元的兴奋性过高和神经元可塑性。在旋毛虫感染的豚鼠(8000条幼虫或生理盐水)中进行了电生理学、免疫学、分子生物学或免疫化学研究,感染时间为急性炎症期(7天)或感染后35天(肠道清除后)。急性炎症导致AH细胞兴奋性过高,并使黏膜和神经组织中的髓过氧化物酶、肥大细胞类胰蛋白酶、前列腺素E2、白三烯B4、脂质过氧化、一氧化氮和明胶酶水平升高;感染后35天仍存在较低水平的炎症。急性暴露于AC、组胺、环氧化酶或白三烯途径的阻滞剂可可逆地抑制AH细胞兴奋性过高。旋毛虫感染后,离体肌间神经节中基础cAMP反应或由福斯高林(FSK)、Ro-20-1724、组胺或P物质诱发的反应增强;钙结合蛋白(AH)神经元中的AC表达和AC免疫反应性也上调。在AH神经元中,通过G蛋白偶联受体(GPCR)作用的组胺(肥大细胞介质)或P物质(神经递质)引起的cAMP依赖性慢兴奋性突触传递样反应,在旋毛虫感染后增强了2.5倍。FSK、组胺、P物质或旋毛虫急性感染导致离体神经节或钙结合蛋白(AH)神经元中cAMP依赖性核CREB磷酸化增加5至30倍。感染后35天,AC和CREB磷酸化水平仍升高。持续的免疫激活、AC上调、磷酸二酯酶IV活性增强以及GPCR-AC/cAMP/pCREB信号通路的易化作用,促成了旋毛虫诱导的神经元可塑性和AH细胞兴奋性过高。这可能与肠道线虫感染和炎症性肠病有关,并且是一个潜在的治疗靶点。