牦牛肺脏气道肺神经内分泌细胞及其相关因子的分布和表达分析

doi:10.11843/j.issn.0366-6964.2025.11.044

摘要/Abstract

摘要：

肺神经内分泌细胞(PNEC)位于气道上皮层内，可分泌活性胺和肽类物质，兼有神经细胞及内分泌细胞的特征，与陆生哺乳动物肺脏发育和适应外界环境功能密切相关。为探究PNEC在不同年龄牦牛肺脏中的分布和低氧适应性结构形成过程中的可能作用，本研究选取初生(1~6日龄)、幼年(1~3岁)、成年(4~6岁)及老年(7~10岁)牦牛各6头，利用Grimelius嗜银染色、免疫组织化学染色、免疫荧光染色对PNEC及其相关因子嗜铬粒蛋白A(CGA)、S100蛋白(S100 A1)、神经元特异性烯醇化酶(NSE)和神经源性分化因子(NEUROD1)的分布特征进行研究，同时采用Western blot和qRT-PCR检测这些相关因子在蛋白水平的表达及基因水平的转录。结果显示，牦牛肺脏气道上皮中PNEC数量以初生组最多；PNEC相关因子主要分布于气道上皮细胞、Ⅱ型肺泡上皮细胞、肺动脉内皮细胞及气管腺腺上皮细胞；CGA、S100A1和NEUROD1的表达趋势基本一致，均在初生组牦牛肺脏中表达最高，之后呈递减趋势，蛋白水平上的表达各组差异均显著(P < 0.05)。基因水平上的转录各组趋势虽然一致，但4种因子组间差异较大，NSE在成年组牦牛肺脏中表达最高，且显著高于初生组、幼年组、老年组牦牛，呈现先递增再递减的趋势。本研究表明PNEC及其相关因子通过动态表达调控可能参与了牦牛肺脏对低氧环境的适应性调节，同时为牦牛生存繁衍提供关键能量支持，也为进一步研究牦牛肺脏低氧适应机制提供了资料。

关键词: 牦牛, 肺, 神经内分泌细胞

Abstract:

Lung neuroendocrine cells (PNEC) are located in the airway epithelium, which can secrete active amines and peptides, and have the characteristics of both nerve cells and endocrine cells, which are closely related to the lung development and adaptation to the external environment of terrestrial mammals. In order to explore the distribution of PNEC in the lungs of yaks of different ages and its possible role in the formation of hypoxic adaptive structure, six yaks were selected from newborn (1-6 days old), juvenile (1-3 years old), adult (4-6 years old) and elderly (7-10 years old). Grimelius silver staining, immunohistochemical staining and immunofluorescence staining were used to study the distribution characteristics of PNEC and its related factors, such as chromogranin A(CGA), S100, neuron-specific enolase (NSE) and neurogenic differentiation factor (NEUROD1). At the same time, Western blot and qRT-PCR were used to detect the expression of these related factors at the protein level and the transcription at the gene level. The results showed that the number of PNEC in the airway epithelium of yak lung was the highest in the newborn group. PNEC-related factors are mainly distributed in airway epithelial cells, type Ⅱ alveolar epithelial cells, pulmonary artery endothelial cells and tracheal glandular epithelial cells. The expression trends of CGA, S100A1 and NEUROD1 were basically the same, with the highest expression in the lungs of newborn yaks, and then showed a decreasing trend, with significant differences in protein levels among groups (P < 0.05). Although the transcription trend of each group at the gene level is the same, there are great differences among the four factors. The expression of NSE in the lungs of adult yaks is the highest, and it is significantly higher than that of newborn yaks, young yaks and old yaks, showing a trend of increasing first and then decreasing. This study shows that PNEC and its related factors may participate in the adaptive regulation of yak lungs to hypoxia environment through dynamic expression regulation, and provide key energy support for yak survival and reproduction, which provides data for further study on the adaptive mechanism of yak lungs to hypoxia.

Key words: yak, lung, neuroendocrine cell

中图分类号:

S852.1

豆婉婉, 崔燕, 张倩, 牛悦悦, 何俊峰. 牦牛肺脏气道肺神经内分泌细胞及其相关因子的分布和表达分析[J]. 畜牧兽医学报, 2025, 56(11): 5888-5900.

DOU Wanwan, CUI Yan, ZHANG Qian, NIU Yueyue, HE Junfeng. Study on the Distribution and Expression of Lung Neuroendocrine Cells and Their Related Factors in Yak Lung Airway[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5888-5900.

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基因名称 Gene name	GenBank登录号 GenBank accession number	引物序列(5′→3′) Primer sequence	片段长度/bp Fragment size
NSE	XM_005907488.1	F: GGTCCAAGTTCACAGCCAATG R: ACCTTCAGCAGCAGACAGTT	121
S100A1	XM_014483048.1	F: AGACTCTCATCAATGTGTTCCA R: CCTCTCCATCTCCGTTCTCA	180
CGA	XM_005902885.1	F: CCAAGAACATCACCTCGGAAG R: GAGCAAGTCAGCCATCATCAG	171
NEUROD1	XM_005905941.2	F: GAGGAGCACGAGACAGACA R: CGCCTTCGTCATCTTCTTCTT	189
β-actin	XM_005887322.2	F: CCGTGACATCAAGGAGAAG R: AGGAAGGAAGGCTGGAAG	174

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