猪流行性腹泻病毒感染对小肠杯状细胞的影响及其机制的初步分析  

Preliminary Study on the Mechanism of Porcine Epidemic Diarrhea Virus Infection Affecting Small Intestinal Goblet Cells

作　　者：刘芮伶 李昱辰 汤荣锋 杨倩[1] LIU Ruiling;LI Yuchen;TANG Rongfeng;YANG Qian(Animal Mucosal Immunology Laboratory,College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,动物黏膜免疫学实验室,南京210095

出　　处：《畜牧兽医学报》2025年第2期900-911,共12页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划项目(2022YFD1800805-02)。

摘　　要：本研究旨在探明猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)对仔猪肠道黏液分泌的影响并揭示其潜在的分子机制。构建仔猪PEDV感染模型,通过阿利新蓝-过碘酸-雪夫(Alcian blue-periodic acid-Schiff,AB-PAS)染色和免疫荧光染色,揭示病毒感染对仔猪肠道杯状细胞数量和功能的影响。进一步利用肠道干细胞构建杯状细胞的体外实验模型,探明PEDV在该细胞模型的复制特点后,初步解析PEDV感染对杯状细胞黏液蛋白转录及其分泌调控相关信号通路的影响。体内试验结果显示,PEDV主要感染仔猪的空肠和回肠,其对空肠具有更高的易感性,并能显著抑制肠道黏膜中杯状细胞的数量和分泌功能。进一步利用肠道干细胞建立了杯状细胞体外培养模型,发现病毒感染显著降低了杯状细胞Muc2、TFF 3和SPDEF基因的转录水平,同时,黏液分泌调控的关键通路MAPK信号通路的活性也受到了明显抑制。PEDV感染会导致仔猪肠道杯状细胞数量减少,黏蛋白分泌水平降低。病毒对MAPK信号途径的抑制可能是导致这一现象的关键原因。The purpose of this study was to find out the specific effects of porcine epidemic diarrhea virus(PEDV)on intestinal goblet cells in piglets,and to reveal its potential mechanism.The model of PEDV infection in piglets was established,and the specific effects of virus infection on the number of goblet cells and mucus secretion in piglets′intestines were revealed by using Alcian blue-periodic acid-Schiff(AB-PAS)staining and immunofluorescence staining.In order to verify the results of in vivo experiments,an in vitro experimental model of goblet cells was further constructed by using intestinal stem cells.In this model,firstly,the replication characteristics of PEDV are explored.Then,the effects of PEDV infection on transcription of functional genes(Muc2,TFF3,SPDEF)in goblet cells were studied.Finally,the influence of virus infection on the core regulatory pathway(MAPK signaling pathway)in the biological process of goblet cells was detected.Through the observation of tissue sections,it was found that PEDV mainly infected the jejunum and ileum of piglets,and jejunum was more susceptible to virus.Infection leads to severe atrophy of intestinal villi,significant decrease of goblet cells and serious loss of mucin.In addition,the in vitro culture model of goblet cells was successfully established,and the replication characteristics of PEDV in this model were revealed.Further research based on this in vitro infection model showed that PEDV infection significantly reduced the transcription level of Muc2,TFF3 and SPDEF genes in goblet cells,and at the same time,the activity of MAPK signaling pathway was also inhibited.PEDV infection can lead to the decrease of goblet cells and mucin secretion level in piglets.The inhibition of MAPK signaling pathway by virus may be the key reason for this phenomenon.

关 键 词：PEDV 杯状细胞 MAPK信号通路 

分 类 号：S858.285.3[农业科学—临床兽医学]