水通道蛋白AQP1、3、4、5、8和9在双峰驼肺中的表达

发布时间：2018-02-28 02:12

本文关键词： 水通道蛋白双峰驼肺免疫组化　出处：《兰州大学》2015年硕士论文　论文类型：学位论文

【摘要】：众所周知,水分乃生命体的重要构成,系生命体存活的基本条件,是生命体最依赖的代谢资源。而长期生活在干旱、沙漠化以及极其缺水的恶劣环境中的双峰驼,却展现出极佳的环境适应性,研究表明,在长期的进化过程中,双峰驼形成了许多不同于其它动物的生理特性,且正因这些独特的生理特性,给予其对沙漠化缺水环境的超强适应力,在这些独特生理特性中,其中呼吸系统中的水分代谢、表面活性物质分泌以及干冷气体加温、加湿等生理过程对其生存适应尤为重要。目的：研究双峰驼呼吸道形态学特征以及水通道蛋白(AQPs)在其肺中的表达情况,探讨双峰驼适应极干旱荒漠环境的呼吸生理机制,以便为更深入地探讨双峰驼为何能如此好的适应极干旱缺水的极恶劣环境,提供相关的呼吸生理代谢机制的形态学和分子生物学基础资料。方法：利用常规形态学统计方法、扫描电子显微镜和石蜡切片HE染色法对双峰驼呼吸道上皮组织形态结构进行统计与分析,利用免疫组化对双峰驼肺组织中AQPs的表达情况进行定位分析以及运用IPP软件对AQPs表达量进行半定量分析。结果：形态学统计表明,双峰驼鼻腔长度较长且多褶皱,鼻腔粘膜富含微血管和腺体细胞,气管长且弯曲,呈大S型,相比于黄牛,肺较致密且含水量高。免疫组化检测显示,在双峰驼呼吸道上皮细胞中有六种水通道蛋白(AQP)表达。其中,AQP1主要表达于呼吸道毛细血管网、淋巴管、胸膜脏层的间皮细胞以及气管上皮细胞顶膜面；AQP3存在于呼吸道上皮表层与腺体的基底细胞膜上；AQP4分布于整个呼吸道上皮杯状细胞基底侧细胞膜和鼻腔粘膜腺体细胞顶膜面；AQP5表达于呼吸道粘膜下腺腺体细胞管腔面和Ⅰ型肺泡上皮细胞膜上；AQP8主要表达于支气管粘膜下腺腺体细胞基底层细胞膜上；AQP9则位于呼吸道上皮细胞。同时IPP软件半定量分析结果表明,双峰驼肺中各种AQP表达情况为SAQP4SAQP5SAQP9SAQP8SAQP1SAQP3,IODAQP4IODAQP5IODAQP8 IODAQP9IODAQP1IODAQP3.结论：呼吸道组织形态学特征证明双峰驼对干旱沙漠环境的适应性,AQPs在双峰驼呼吸道上皮细胞的强烈表达,与嗅觉敏感性、气道润化、气道水平衡、新生血管形成、气道表面液体层、气道反应性以及肺内液体转运等有关,AQPs在双峰驼呼吸道上皮中的高度表达,为其适应极干旱荒漠环境提供了蛋白水平依据。
[Abstract]:As we all know, water is an important component of life, a basic condition for its survival, and a metabolic resource on which it is most dependent. The Bactrian camel, which has lived for a long time in harsh environments of drought, desertification and extreme water scarcity, But showing excellent environmental adaptation, studies have shown that over the course of long evolution, Bactrian camels have developed many physiological characteristics different from those of other animals, and because of these unique physiological characteristics, In these unique physiological characteristics, including water metabolism in the respiratory system, the secretion of surface active substances and the warming of dry and cold gases, Humidification and other physiological processes are particularly important for their survival and adaptation. Objective: to study the respiratory morphological characteristics and the expression of aquaporin aquaporin in the lungs of Bactrian Camels, and to explore the respiratory physiological mechanism of Bactrian camel adapting to extremely arid desert environment. In order to further study why Bactrian camel can adapt to the extremely harsh environment of extreme drought and water shortage, and provide the basic information of morphology and molecular biology about the mechanism of respiratory physiological metabolism. Methods: using conventional morphological statistical method, The morphology and structure of respiratory epithelium of Bactrian camel were analyzed by scanning electron microscope and paraffin section HE staining. The expression of AQPs in the lung tissue of bactrian camel was analyzed by immunohistochemistry and the expression of AQPs was semi-quantitatively analyzed by IPP software. Results: morphological statistics showed that the length of nasal cavity of Bactrian camel was long and pleated. The nasal mucosa is rich in microvascular and glandular cells. The trachea is long and curved. Compared with cattle, the lungs are denser and have higher water content. Six aquaporins (aquaporins) are expressed in the epithelial cells of the respiratory tract of Bactrian camel. The mesothelial cells of pleural layer and the apical membrane of tracheal epithelial cells AQP3 exist on the basal cell membrane of respiratory epithelial surface and gland. AQP4 is distributed in the basal cell membrane of the whole respiratory epithelium goblet cell and the fine gland of nasal cavity mucosa of the whole respiratory epithelium goblet cell. AQP5 was expressed on the submucosal glandular cells of respiratory tract and on the membrane of type I alveolar epithelium. AQP9 was mainly expressed on the basal cell membrane of glandular cells in submucous membrane of the bronchus, whereas AQP9 was located in the epithelial cells of the respiratory tract. The results of semi-quantitative analysis by IPP software show that, The expression of SAQP4SAQP5SAQP9SAQP8SAQP1SAQP3ODAQP4IODAQP5IODAQP8 IODAQP9IODAQP1IODAQP3 in the lungs of bactrian camels was found to be strongly expressed in the respiratory epithelium of Bactrian camel. Conclusion: the morphological features of respiratory tract tissue indicate the adaptability of bactrian camel to arid desert environment and its strong expression, olfactory sensitivity, airway moisturization. Airway water balance, neovascularization, airway surface liquid layer, airway reactivity and lung fluid transport are related to the high expression of AQPs in respiratory epithelium of Bactrian Camel, which provides a basis for protein level in adapting to extremely arid desert environment.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S824;Q78

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