血管活性肠肽、降钙素基因相关肽及其受体在气道高反应动物肺内的时空分布(英文)  

Temporal and spatial distribution of VIP, CGRP and their receptors in the development of airway hyperresponsiveness in the lungs

作　　者：任雁宏[1] 秦晓群[1] 管茶香[1] 罗自强[1] 张长青[1] 孙秀泓[1] 

机构地区：[1]中南大学湘雅医学院生理学教研室,长沙410078

出　　处：《生理学报》2004年第2期137-146,共10页Acta Physiologica Sinica

基　　金：This work was supported by the National Natural Science Foundation of China (No. 30270586).

摘　　要：为探讨内源性神经肽在气道高反应性形成中的作用,我们以臭氧应激损伤动物气道上皮细胞,建立气道高反应性动物模型,并观察臭氧应激不同时间肺内血管活性肠肽(vasoactive intestinal peptide,VIP)、降钙素基因相关肽(calcitonin gene-related peptide,CGRP)含量变化以及VIP受体(VIPR1)、CGRP受体(GRPR1)mRNA在肺内表达、分布的改变。实验观察到,臭氧应激组动物吸入乙酰甲胆碱后气道阻力高于正常对照组,肺内呈现明显的炎症改变。随臭氧应激时间延长,肺组织匀浆中VIP、CGRP浓度呈先增高后降低的双向改变,CGRP达峰值时间早于VIP。VIPR1、CGRPR1 mRNA表达亦经历了双向过程,VIPR1峰值持续时间长于CGRPR1。在无应激对照组动物,肺间质、支气管上皮细胞、血管内皮细胞、平滑肌细胞均有VIPR1、CGRPR1 mRNA表达。随应激时间的延长,阳性细胞呈斑片状集中于气管、血管周围,染色强度增加,至臭氧应激第8天,阳性染色细胞减少。因此我们推测,臭 氧应激可以诱导动物气道高反应性的形成。在炎症的早期,以CGRP的作用为主,与肺损伤早期炎症信号的传递有关,以清除刺激 原、及时终止致病原的作用;炎症后期以保护机体、促进修复、减轻损伤为主,VIP发挥主要作用。To explore the role of intrapulmonary neuropeptides in the development of airway hyperresponsiveness, we established an animal model of airway hyperresponsiveness (AHR) in rabbits by using ozone exposure. With the model, after test of the mechanics of respiration and bronchoalveolar lavage assay, the levels of vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) in the lungs were determined by radioimmunoassay, and the expression of mRNA coding receptors of these two neuropeptides was evaluated by reverse transcriptional-polymerase chain reaction (RT-PCR). At the same time, the distribution of VIP receptor-1 (VIPR1) and CGRP receptor-1 (CGRPR1) in lung tissues and its time-course were examined by in situ hybridization. The results showed: (1) in ozone-stressing groups, airway resistance increased significantly and typical inflammatory pathological changes were observed in pulmonary tissue slides, including neutrophil and eosinophil infiltration, mucus exudation and bronchial epithelial cells (BECs) shedding; (2) with elongation of ozone exposure, the levels of VIP and CGRP in the lungs increased at first, reaching a peak on d 2 to 4, then decreased slowly, and CGRP peaked somewhat earlier than VIP; (3) mRNA expression of the two neuropeptide receptors in the lungs changed in a similar manner like VIP and CGRP, but the high level of mRNA expression of VIPR1 lasted longer than that of CGRPR1; and (4) in situ hybridization for neuropeptide receptors demonstrated that, in unstressed control, VIPR1 and CGRPR1 positive cells appeared in the airway epithelium, pulmonary interstitial and focal areas of airway and vascular smooth muscles. With the elongation of ozone exposure, hybridization stained deeper and the majority of positive cells were located around the vessels and bronchus except a few in the alveoli. At 8 d, only a small number of positive cells were seen in the lungs. From the results, it is concluded that ozone-stressing can induce the development of AHR, in which VIP and CGRP may p

关 键 词：气道高反应 动物模型 支气管上皮细胞 神经肽 

分 类 号：R56[医药卫生—呼吸系统]