Article(id=1307007644934361978, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, articleNumber=null, orderNo=null, doi=10.11843/j.issn.0366-6964.2026.01.046, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1744041600000, receivedDateStr=2025-04-08, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1789544505580, onlineDateStr=2026-09-16, pubDate=1769097600000, pubDateStr=2026-01-23, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789544505580, onlineIssueDateStr=2026-09-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789544505580, creator=13701087609, updateTime=1789544505580, updator=13701087609, issue=Issue{id=1307007396681900041, tenantId=1146029695717560320, journalId=1306274286668972124, year='2026', volume='57', issue='1', pageStart='1', pageEnd='568', issueExtLink='null', onlineDate='null', pubDate='1769097600000', pubDateStr='2026-01-23', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789544446392, creator='13701087609', updateTime=1789609890784, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1307281890638057732, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1307281890738721029, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307007396681900041, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=526, endPage=539, ext={EN=ArticleExt(id=1307007646226207613, articleId=1307007644934361978, tenantId=1146029695717560320, journalId=1306274286668972124, language=EN, title=Effect of Radix dichroa Powder on Intestinal Mechanical Barrier of Chicks Infected with Coccidia at Different Developmental Stages, columnId=1307007468660355587, journalTitle=Acta Veterinaria et Zootechnica Sinica, columnName=CLINICAL VETERINARY MEDICINE, runingTitle=null, highlight=null, articleAbstract=

The aim of this study was to investigate the effect of Radix dichroa powder (RDP) on intestinal mechanical barrier at different developmental stages of chicks infected with coccidia. The experiment was divided into 6 groups, which were administered on the day before the challenge, the day of the attack, and the 2nd, 3rd, 4th and 5th days after the attack.Each experimental group included Group I (Healthy control group), Group Ⅱ (Infection control group) and Group Ⅲ (RDP recommended dose group). At the end of the administration, the chick cecum of each group were collected to make pathological sections. The immunohistochemical analysis of intestinal tight junction proteins occludin and claudin-1 were performed by Pro PLUS 6.0 image software, and the secretion of intestinal goblet cells and mucin were determined.The results showed that compared with Group Ⅰ, the expression levels of occludin protein, claudin-1 protein, and intestinal mucin in Group Ⅱ and Group Ⅲ were significantly reduced (P<0.05), and the number of goblet cells was significantly decreased (P<0.05). Compared with Group Ⅱ, the expression of the above three proteins of Group Ⅲ were significantly improved (P<0.05), and the number of goblet cells were increased from the day before the challenge to 3 days after the challenge. However, with the continuous development of coccidia in chicks and the delay in administration time, the above three proteins in Group Ⅱ and Group Ⅲ were markedly decreased on the 4th and 5th day of challenge (P>0.05), and the goblet cells disapeared.In summary, RDP can reduce the damage of coccidia to the host intestinal tract, improve the secretion of intestinal barrier protein, and reduce the damage of coccidia to the intestinal structure; the earlier the drug is administered after the challenge, the better the improvement of intestinal damage caused by coccidia.

, authors=Zhiting GUO1, 2, Huanhuan SUN1, Shuning LIU1, Jianxi LI2, Shaobo ZHANG2, Yuan LIU2, Xiaoqin LUO2, Xiaocheng WEI1, Chengyi LI1, authorsList=Zhiting GUO, Huanhuan SUN, Shuning LIU, Jianxi LI, Shaobo ZHANG, Yuan LIU, Xiaoqin LUO, Xiaocheng WEI, Chengyi LI, authorCompany=null, correspAuthors=Chengyi LI, authorNote=null, correspAuthorsNote=
LI Chengyi, E-mail:
, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1307007656145736592, articleId=1307007644934361978, tenantId=1146029695717560320, journalId=1306274286668972124, language=CN, title=常山散对感染不同发育阶段球虫雏鸡盲肠机械屏障的影响, columnId=1307007468815544837, journalTitle=畜牧兽医学报, columnName=临床兽医, runingTitle=null, highlight=null, articleAbstract=

旨在研究常山散对感染球虫不同发育阶段雏鸡肠道机械屏障的影响。试验分为6组,分别为攻虫前1天、攻虫当天,攻虫后第2、3、4和5天进行给药,每个试验组包括Ⅰ组(健康对照组)、Ⅱ组(感染对照组)和Ⅲ组(常山散推荐剂量组)。给药结束采集各组鸡盲肠制作病理切片,应用Image Pro PLUS 6.0图像软件对肠紧密连接蛋白Occludin和Claudin-1进行免疫组化分析,测定肠道杯状细胞和黏蛋白的分泌情况。结果表明:与Ⅰ组相比,Ⅱ组和Ⅲ组Occludin蛋白、Claudin-1蛋白和肠黏蛋白表达量显著降低(P<0.05),杯状细胞数显著减少(P<0.05)。与Ⅱ组相比,攻虫前1天至攻虫后第3天给药,Ⅲ组以上三种蛋白表达量明显升高(P<0.05),杯状细胞数增多;但随着球虫在鸡体内不断发育和给药时间的推迟,攻虫后第4和5天给药Ⅱ组和Ⅲ组以上3种蛋白表达量很少(P>0.05),杯状细胞消失。综上,常山散可减轻球虫对宿主肠道的破坏,促进肠道屏障蛋白分泌,减轻球虫对肠道结构的损害;且攻虫后越早给药,对球虫造成的肠道损伤改善效果越好。

, authors=郭志廷1, 2, 孙焕焕1, 刘淑宁1, 李建喜2, 张少博2, 刘圆2, 罗晓琴2, 魏小成1, 李成义1, authorsList=郭志廷, 孙焕焕, 刘淑宁, 李建喜, 张少博, 刘圆, 罗晓琴, 魏小成, 李成义, authorCompany=null, correspAuthors=李成义, authorNote=

郭志廷,博士生,主要从事新兽药研发和中药药理学研究,E-mail:

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李成义,主要从事中药鉴定与品质评价研究,E-mail:
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Agricultural Sciences,Lanzhou 730050,China), AuthorCompanyExt(id=1307286474743894644, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, companyId=1307286474722923122, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.中国农业科学院兰州畜牧与兽药研究所,农业农村部兽用药物创制 重点实验室,甘肃省中兽医技术创新中心,兰州 730050)])], figs=[ArticleFig(id=1307286477986091693, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.1, caption=Immunohistochemical images of Occludin protein in the cecum of chicks in each experimental group

The red arrow indicates the Occludin protein at the top of the intestinal mucosal cell membrane, and the blue arrow indicates the absence of Occludin protein. Bar=100 μm

, figureFileSmall=0314V0EXL/Q++NvUwVNREg==, figureFileBig=ggK+5EW0SsxRa2QPHUmO6Q==, tableContent=null), ArticleFig(id=1307286478069977774, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图1, caption=各试验组雏鸡盲肠Occludin蛋白免疫组化图片

红色箭头指示肠黏膜细胞膜顶端Occludin蛋白部分,蓝色箭头指示无Occludin蛋白部分。标尺=100 μm

, figureFileSmall=0314V0EXL/Q++NvUwVNREg==, figureFileBig=ggK+5EW0SsxRa2QPHUmO6Q==, tableContent=null), ArticleFig(id=1307286478179029679, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.2, caption=Mean optical density values of Occludin protein immunohistochemically positive area in the cecum of chicks in each experimental group

A-F. Dosing 1 day, dosing on the day, dosing on the 2nd, 3rd, 4th, 5th day of attack. The same as below. Compared with the healthy group, *.P<0.05; Compared with the infection group, #.P<0.05

, figureFileSmall=a45I8VpSDZK1r+Id0qSByw==, figureFileBig=e6vjipD4tMXE4ENco0r9OA==, tableContent=null), ArticleFig(id=1307286478254527152, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图2, caption=各试验组雏鸡盲肠Occludin蛋白免疫组化阳性面积平均光密度值

A~F.攻虫前1天、攻虫当天、攻虫第2、3、4、5天给药。下同。与健康组相比,*.P<0.05;与感染组相比,#.P<0.05

, figureFileSmall=a45I8VpSDZK1r+Id0qSByw==, figureFileBig=e6vjipD4tMXE4ENco0r9OA==, tableContent=null), ArticleFig(id=1307286478321636017, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.3, caption=Immunohistochemical images of Claudin-1 protein in the cecum of chicks in each experimental group

The red arrow indicates the Claudin-1 protein at the top of the intestinal mucosal cell membrane, and the blue arrow indicates the absence of Claudin-1 protein. Bar=100 μm

, figureFileSmall=tJrPzJuwMmvPjn8QdidODQ==, figureFileBig=9i3U4r+mszsfKNMJjcwQuA==, tableContent=null), ArticleFig(id=1307286478397133490, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图3, caption=各试验组雏鸡盲肠Claudin-1蛋白免疫组化图片

红色箭头指示肠黏膜细胞膜顶端Claudin-1蛋白部分,蓝色箭头指示无Claudin-1蛋白部分。标尺=100 μm

, figureFileSmall=tJrPzJuwMmvPjn8QdidODQ==, figureFileBig=9i3U4r+mszsfKNMJjcwQuA==, tableContent=null), ArticleFig(id=1307286478464242355, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.4, caption=Mean optical density values of Claudin-1 protein immunohistochemically positive area in the cecum of chicks in each experimental group

Compared with the healthy group, *. P<0.05; Compared with the infection group, #. P<0.05

, figureFileSmall=DSOvtcivhHJAox/MvyvMWA==, figureFileBig=6/WPSCol2MpCjcvwNML/dw==, tableContent=null), ArticleFig(id=1307286478543934132, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图4, caption=各试验组雏鸡盲肠Claudin-1蛋白免疫组化阳性面积平均光密度值

与健康组相比,*. P<0.05;与感染组相比,#. P<0.05

, figureFileSmall=DSOvtcivhHJAox/MvyvMWA==, figureFileBig=6/WPSCol2MpCjcvwNML/dw==, tableContent=null), ArticleFig(id=1307286478627820213, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.5, caption=AB-PAS staining image of cecal mucin in each experimental group

The black arrow indicates mucin. Bar=100 μm

, figureFileSmall=KmA91ZyZv0SW+dHlrINxNQ==, figureFileBig=P/dfKwurMoa2Ccyy9msKpg==, tableContent=null), ArticleFig(id=1307286478720094902, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图5, caption=各试验组雏鸡盲肠黏蛋白AB-PAS染色图片

黑色箭头指示黏蛋白。标尺=100 μm

, figureFileSmall=KmA91ZyZv0SW+dHlrINxNQ==, figureFileBig=P/dfKwurMoa2Ccyy9msKpg==, tableContent=null), ArticleFig(id=1307286478791398071, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.6, caption=Mean optical density values of mucin-positive areas in the chickcecum of in each experimental group

Compared with the healthy group, *. P<0.05; Compared with the infection group, #. P<0.05

, figureFileSmall=BIMhwDkELtZ0ONmel6SfEQ==, figureFileBig=Els2bRXkT+8piyoSne9qxQ==, tableContent=null), ArticleFig(id=1307286478858506936, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图6, caption=各试验组雏鸡盲肠黏蛋白阳性面积平均光密度值

与健康组相比,*. P<0.05;与感染组相比,#. P<0.05;

, figureFileSmall=BIMhwDkELtZ0ONmel6SfEQ==, figureFileBig=Els2bRXkT+8piyoSne9qxQ==, tableContent=null), ArticleFig(id=1307286478938198713, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.7, caption=PAS staining images of chick cecal goblet cells in each experimental group

Red arrows indicates goblet cells. Bar=100 μm

, figureFileSmall=xtw7KXKekemX81DaRDKEDw==, figureFileBig=U404pphcfucq9fe3GcO2Hg==, tableContent=null), ArticleFig(id=1307286479009501882, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图7, caption=各试验组雏鸡盲肠杯状细胞PAS染色图片

红色箭头指示杯状细胞。标尺=100 μm

, figureFileSmall=xtw7KXKekemX81DaRDKEDw==, figureFileBig=U404pphcfucq9fe3GcO2Hg==, tableContent=null), ArticleFig(id=1307286479068222139, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Fig.8, caption=Number of goblet cells in the chick cecum in each experimental group

Compared with the healthy group, *.P<0.05; Compared with the infection group, #.P<0.05

, figureFileSmall=DCxq8Ha5QfKo+XVo8jyQuQ==, figureFileBig=dETM9pDrKgrt4JW9OobSPA==, tableContent=null), ArticleFig(id=1307286479126942396, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=图8, caption=各试验组雏鸡盲肠杯状细胞数量

与健康组相比,*.P<0.05;与感染组相比,#.P<0.05

, figureFileSmall=DCxq8Ha5QfKo+XVo8jyQuQ==, figureFileBig=dETM9pDrKgrt4JW9OobSPA==, tableContent=null), ArticleFig(id=1307286479189856957, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=EN, label=Table 1, caption=

Grouping and treatment of test chicks

, figureFileSmall=null, figureFileBig=null, tableContent=

项目

Item

组别

Group

动物数

Animal numbers

子孢子接种量

Subspore inoculum volume

饲料中药物浓度

Drug concentration in feed

给药时间/d

Days of administration

攻虫前1天给药

Dosing 1d before attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫当天给药

Dosing on the day of attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第2天给药

Dosing on the 2nd day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第3天给药

Dosing on the 3d day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第4天给药

Dosing on the 4th day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第5天给药

Dosing on the 5th day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d
), ArticleFig(id=1307286479252771518, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307007644934361978, language=CN, label=表1, caption=

试验鸡分组与处理

, figureFileSmall=null, figureFileBig=null, tableContent=

项目

Item

组别

Group

动物数

Animal numbers

子孢子接种量

Subspore inoculum volume

饲料中药物浓度

Drug concentration in feed

给药时间/d

Days of administration

攻虫前1天给药

Dosing 1d before attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫当天给药

Dosing on the day of attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第2天给药

Dosing on the 2nd day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第3天给药

Dosing on the 3d day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第4天给药

Dosing on the 4th day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d

攻虫后第5天给药

Dosing on the 5th day after the attack

Ⅰ10———
Ⅱ105×104·只-1——
Ⅲ105×104·只-1常山散(0.1 g·kg-1)连续给药4 d
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郭志廷 1, 2 , 孙焕焕 1 , 刘淑宁 1 , 李建喜 2 , 张少博 2 , 刘圆 2 , 罗晓琴 2 , 魏小成 1 , 李成义 1
畜牧兽医学报 | 临床兽医 2026,57(1): 526-539
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畜牧兽医学报 |临床兽医 2026 , 57 (1) : 526 -539
常山散对感染不同发育阶段球虫雏鸡盲肠机械屏障的影响
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郭志廷1, 2 , 孙焕焕1, 刘淑宁1, 李建喜2, 张少博2, 刘圆2, 罗晓琴2, 魏小成1, 李成义1
作者信息
  • 1.甘肃中医药大学药学院,兰州 730000
  • 2.中国农业科学院兰州畜牧与兽药研究所,农业农村部兽用药物创制 重点实验室,甘肃省中兽医技术创新中心,兰州 730050
通讯作者:
李成义,主要从事中药鉴定与品质评价研究,E-mail:
作者简介:

郭志廷,博士生,主要从事新兽药研发和中药药理学研究,E-mail:

Effect of Radix dichroa Powder on Intestinal Mechanical Barrier of Chicks Infected with Coccidia at Different Developmental Stages
Zhiting GUO1, 2 , Huanhuan SUN1, Shuning LIU1, Jianxi LI2, Shaobo ZHANG2, Yuan LIU2, Xiaoqin LUO2, Xiaocheng WEI1, Chengyi LI1
Affiliations
  • 1.College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou,Gansu Provicne 730000,China
  • 2.Key Lab of Veterinary Pharmaceutical Development,Ministry of Agriculture and Rural Affairs & Technology Innovation Center of Traditional Chinese Veterinary,Gansu Province,Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China
  • Corresponding Author:
    LI Chengyi, E-mail:
出版时间: 2026-01-23 doi: 10.11843/j.issn.0366-6964.2026.01.046
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旨在研究常山散对感染球虫不同发育阶段雏鸡肠道机械屏障的影响。试验分为6组,分别为攻虫前1天、攻虫当天,攻虫后第2、3、4和5天进行给药,每个试验组包括Ⅰ组(健康对照组)、Ⅱ组(感染对照组)和Ⅲ组(常山散推荐剂量组)。给药结束采集各组鸡盲肠制作病理切片,应用Image Pro PLUS 6.0图像软件对肠紧密连接蛋白Occludin和Claudin-1进行免疫组化分析,测定肠道杯状细胞和黏蛋白的分泌情况。结果表明:与Ⅰ组相比,Ⅱ组和Ⅲ组Occludin蛋白、Claudin-1蛋白和肠黏蛋白表达量显著降低(P<0.05),杯状细胞数显著减少(P<0.05)。与Ⅱ组相比,攻虫前1天至攻虫后第3天给药,Ⅲ组以上三种蛋白表达量明显升高(P<0.05),杯状细胞数增多;但随着球虫在鸡体内不断发育和给药时间的推迟,攻虫后第4和5天给药Ⅱ组和Ⅲ组以上3种蛋白表达量很少(P>0.05),杯状细胞消失。综上,常山散可减轻球虫对宿主肠道的破坏,促进肠道屏障蛋白分泌,减轻球虫对肠道结构的损害;且攻虫后越早给药,对球虫造成的肠道损伤改善效果越好。

常山散  /  雏鸡  /  球虫  /  肠道  /  机械屏障

The aim of this study was to investigate the effect of Radix dichroa powder (RDP) on intestinal mechanical barrier at different developmental stages of chicks infected with coccidia. The experiment was divided into 6 groups, which were administered on the day before the challenge, the day of the attack, and the 2nd, 3rd, 4th and 5th days after the attack.Each experimental group included Group I (Healthy control group), Group Ⅱ (Infection control group) and Group Ⅲ (RDP recommended dose group). At the end of the administration, the chick cecum of each group were collected to make pathological sections. The immunohistochemical analysis of intestinal tight junction proteins occludin and claudin-1 were performed by Pro PLUS 6.0 image software, and the secretion of intestinal goblet cells and mucin were determined.The results showed that compared with Group Ⅰ, the expression levels of occludin protein, claudin-1 protein, and intestinal mucin in Group Ⅱ and Group Ⅲ were significantly reduced (P<0.05), and the number of goblet cells was significantly decreased (P<0.05). Compared with Group Ⅱ, the expression of the above three proteins of Group Ⅲ were significantly improved (P<0.05), and the number of goblet cells were increased from the day before the challenge to 3 days after the challenge. However, with the continuous development of coccidia in chicks and the delay in administration time, the above three proteins in Group Ⅱ and Group Ⅲ were markedly decreased on the 4th and 5th day of challenge (P>0.05), and the goblet cells disapeared.In summary, RDP can reduce the damage of coccidia to the host intestinal tract, improve the secretion of intestinal barrier protein, and reduce the damage of coccidia to the intestinal structure; the earlier the drug is administered after the challenge, the better the improvement of intestinal damage caused by coccidia.

Radix dichroa powder  /  chicks  /  coccidia  /  intestine  /  mechanical barrier
郭志廷, 孙焕焕, 刘淑宁, 李建喜, 张少博, 刘圆, 罗晓琴, 魏小成, 李成义. 常山散对感染不同发育阶段球虫雏鸡盲肠机械屏障的影响. 畜牧兽医学报, 2026 , 57 (1) : 526 -539 . DOI: 10.11843/j.issn.0366-6964.2026.01.046
Zhiting GUO, Huanhuan SUN, Shuning LIU, Jianxi LI, Shaobo ZHANG, Yuan LIU, Xiaoqin LUO, Xiaocheng WEI, Chengyi LI. Effect of Radix dichroa Powder on Intestinal Mechanical Barrier of Chicks Infected with Coccidia at Different Developmental Stages[J]. Acta Veterinaria et Zootechnica Sinica, 2026 , 57 (1) : 526 -539 . DOI: 10.11843/j.issn.0366-6964.2026.01.046
球虫寄生于鸡肠上皮细胞,使肠上皮细胞崩解和坏死,破坏肠黏膜的完整性,而肠黏膜对于维持健康、预防组织损伤和多种疾病,以及向机体提供营养等方面至关重要[1]。柔嫩艾美耳球虫是一种严格的宿主和组织特异性寄生虫,寄生在鸡盲肠隐窝上皮细胞中,裂殖生殖时破坏肠黏膜的完整性,导致盲肠上皮细胞变性、坏死和崩解[2]。紧密连接是肠上皮细胞间的主要连接,其功能是调节肠黏膜屏障的通透性。球虫入侵宿主细胞时,紧密连接蛋白通常是球虫攻击的靶点,导致紧密连接被破坏,肠上皮细胞通透性增加,肠道产生炎症和损伤[3]。闭合蛋白(claudins)和闭锁蛋白(occludin-1)是主要的肠上皮紧密连接蛋白。研究表明,鸡感染球虫后肠黏膜屏障被破坏,Occludin的mRNA表达水平下降,Claudin-1 mRNA表达水平先下降后上升[4]。杯状细胞是肠道特化的黏液分泌细胞,主要合成和释放黏蛋白,形成保护性黏液层[5]。黏蛋白的作用包括:润滑肠壁、阻隔病原体、维持菌群平衡、支持上皮修复及免疫调节[6]。杯状细胞-黏蛋白系统是肠道屏障的核心组成部分,两者协同维护肠道正常屏障功能。
研究表明,中草药微生态制剂通过刺激肠上皮细胞表达黏蛋白,抑制肠道黏膜的炎症反应等方式改善肠道损伤[7]。中药复方“香芪汤”可以减少球虫的增殖,抑制感染鸡肠道组织的炎症和氧化应激反应,维持肠道屏障的完整性[8]。枯草芽胞杆菌复合微生态制剂可缓解坏死性肠炎所致的鸡回肠黏膜形态结构改变,减轻产气荚膜梭菌对回肠黏膜的刺激,提高机体抗氧化水平,减轻机体炎症反应[9]。金银花、紫花地丁和连翘均可保护肠道黏膜屏障功能,阻止D-乳酸、二胺氧化酶和内毒素进入血液循环,还可促进有益菌群繁殖,抑制致病菌群生长[10]。复方青蒿煎剂对雏鸡肠道黏膜免疫无明显效果,但抗炎效果明显,对鸡球虫病具有一定的预防和治疗作用[11]。
中药具有耐药风险低、安全、副作用小等优势,逐渐成为消费者追求的主流产品,部分中药及其提取物还可提高家禽感染球虫病后机体的恢复能力[12⁃14]。中药常山为虎耳草科植物常山(Dichroa febrifuga Lour.)的干燥根,主产于四川、甘肃、江西等地,民间用于治疗疟疾[15]。《神农本草经》中记载常山性寒,味苦辛,有毒,归肺、肝、心经,具有涌吐痰涎、截疟的功效。我国科学家很早就发现常山中主要有效成分为常山碱和异常山碱[16]。现代研究表明,常山碱及其衍生物具有良好的抗疟疾、抗肿瘤、抗球虫、退热、抗炎以及治疗糖尿病、硬皮病等功效,临床上具有很高的科学研究价值[17⁃18]。本试验通过常山散对感染柔嫩艾美耳球虫雏鸡盲肠的Caudin-1、Occludin蛋白免疫组化分析、肠道杯状细胞和黏蛋白分泌的检测,研究对球虫感染雏鸡肠道机械屏障的影响,为常山散科学防治鸡球虫病和药理学研究提供试验依据。
1日龄白羽肉鸡,购自酒泉市旺苗禽业养殖场,前期于SPF隔离舱内饲养至12日龄,后转移至严格消杀且无球虫污染的试验动物房,并于14日龄滴鼻接种鸡新城疫-传染性支气管炎二联苗,饲养至16日龄。
柔嫩艾美耳球虫,由中国农业大学寄生虫病实验室提供。试验开始前,将柔嫩艾美耳球虫卵囊重新进行传代,确保卵囊感染活力。将孢子化率为85%以上的球虫卵囊置于2.5%重铬酸钾溶液4 ℃保存备用。
常山散中试产品(100 g·袋-1,批号2021111501,1 g本品相当于常山原药材10 g;主要有效成分是常山生物碱,含量1.5%),石家庄正道动物药业有限公司生产。常山散主要生产工艺:利用超声波提取技术,从中药常山中提取得到常山浸膏,干燥粉碎后,加入适量淀粉混匀即得(常山浸膏和淀粉质量比9∶1)[19];鸡新城疫-传染性支气管炎二联活疫苗(LaSota株+H120株),购自乾元浩生物股份有限公司南京生物药厂。HRP山羊抗兔IɡG二抗,购自武汉赛维尔生物科技有限公司。PAS染色试剂盒和AB-PAS染色试剂盒购至北京索莱宝科技有限公司。
试验开始前连续3 d进行粪检,确认雏鸡没有感染球虫。试验分为6组:攻虫前1天(15日龄)、攻虫当天(16日龄),攻虫后第2天(17日龄)、第3天(18日龄)、第4天(19日龄)和第5天(20日龄)进行给药。每个试验组包括Ⅰ组(健康对照组)、Ⅱ组(感染性对照组)和Ⅲ组(常山散推荐剂量组)。每组10只鸡,共180只,Ⅰ组单独饲养,各试验组开始试验之前均禁食12 h,正常饮水。除Ⅰ组外,其他各组试验鸡于16日龄按照5×104·只-1鸡人工感染柔嫩艾美耳球虫孢子化卵囊。各试验组鸡连续给药4 d,先称重后捕杀,观察盲肠病理变化,采集鸡盲肠制作病理切片。具体分组见表1。
按文献报道方法进行鸡盲肠免疫组化分析[20]。主要过程:盲肠经福尔马林固定后,石蜡包埋,常规制备石蜡切片,与Occludin和Claudin-1特异性一抗在4 ℃孵育过夜,然后用PBS冲洗,用HRP山羊抗兔IɡG二抗孵育。PBS再次洗涤后,用二氨基联苯胺(DAB)处理,复染,光学显微镜下100倍放大观察。采用Image Pro PLUS 6.0图像分析软件(广州市元奥仪器有限公司)计算3个随机选取区域的平均阳性面积,分析免疫组化试验结果。
盲肠组织样品用4%多聚甲醛固定24 h,梯度乙醇溶液脱水,转入二甲苯,石蜡包埋,切成5 mm厚。切片经脱蜡、复水、放置于水中,根据试剂盒说明书进行PAS染色和AB-PAS染色。光学显微镜下观察黏蛋白和杯状细胞,随机选取三个视野,由Image Pro PLUS 6.0图像分析软件(广州市元奥仪器有限公司)测定杯状细胞和分析黏蛋白阳性面积。
由Image Pro PLUS6.0图像分析软件测量杯状细胞数量,检测免疫组化和黏蛋白阳性密度平均值,试验结果运用SPSS 26.0软件进行统计分析,试验数据比较采用单因素方差分析,使用LSD法进行多重比较,试验数据以“x±s”表示。
Occludin蛋白对维持肠道屏障的通透性具有重要意义。免疫组化法能够将Occludin蛋白染色为深棕色,如图1中红色箭头所示。Occludin蛋白主要围绕肠黏膜细胞顶端分布。在Ⅰ组的鸡盲肠中,Occludin蛋白染色分布密集且均匀;而Ⅱ组的鸡盲肠阳性颗粒染色则因攻虫而明显减少。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药的Ⅲ组中,鸡盲肠阳性颗粒染色继续呈现均匀分布的特征;但与Ⅰ组相比,其阳性颗粒染色密度有所下降。随着球虫在宿主体内的不断发育和给药时间的推迟,攻虫后第4和5天给药组中Ⅱ组和Ⅲ组Occludin蛋白表达无显著变化(P>0.05)。通过对Occludin蛋白染色结果进行定量分析,得到了一致的试验结果,如图2所示。与Ⅰ组相比,Ⅱ组和Ⅲ组Occludin蛋白表达显著下降(P<0.05)。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药显著提升了Occludin蛋白的表达水平(P<0.05),而攻虫后第4天和第5天给药则无显著改善作用。
Claudin-1蛋白反映了肠机械屏障的完整性。免疫组化结果表明,Claudin-1蛋白被染成深棕色阳性颗粒,在Ⅰ组雏鸡盲肠上皮黏膜细胞顶端呈现密集且均匀分布的态势。而在Ⅱ组中,阳性颗粒的密度则明显下降,分布不均匀。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药的Ⅲ组中,雏鸡盲肠阳性颗粒密度则明显增多,且在肠上皮中均匀分布。攻虫后第4和5天给药则不能明显改善阳性颗粒的密度(图3)。随后对免疫组化结果进行定量分析,结果表明与Ⅰ组相比,Ⅱ组和Ⅲ组Claudin-1蛋白表达显著下降(P<0.05)。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药显著提升Claudin-1蛋白的表达水平(P<0.05),而第4和5天给药则无显著改善作用(图4)。
黏蛋白具有润滑肠壁、阻隔病原体和维持菌群平衡等作用。由图5可知,AB-PAS染色雏鸡盲肠结构中黏蛋白被染成蓝色或蓝紫色阳性颗粒。在Ⅰ组雏鸡盲肠上皮黏膜中杯状细胞很多,呈线性分布且染色分布均匀。在Ⅱ组中,阳性染色颗粒的密度明显下降,分布不均匀。由图6可知,与Ⅰ组相比,攻虫前1天给药至攻虫后第5天给药Ⅱ组和Ⅲ组的黏蛋白表达显著降低(P<0.05)。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药Ⅲ组黏蛋白的表达量显著提升(P<0.05),但随着球虫在宿主体内的不断发育和给药时间的推迟,攻虫后第4和5天给药Ⅲ组的黏蛋白表达很少且差异不显著(P>0.05)。
杯状细胞主要合成和释放黏蛋白,形成保护性黏液层。由图7可知,PAS染色雏鸡盲肠结构中杯状细胞被染成紫色阳性颗粒。在Ⅰ组雏鸡盲肠上皮黏膜中杯状很多,呈线性分布且染色分布均匀。在Ⅱ组中,阳性染色颗粒的密度明显下降,分布不均匀。由图8可知,与Ⅰ组相比,攻虫前1天给药至攻虫后第5天给药Ⅱ组和Ⅲ组的杯状细胞显著减少(P<0.05)。与Ⅱ组相比,攻虫前1天给药至攻虫后第3天给药Ⅲ组的杯状细胞数量显著升高(P<0.05),但随着球虫在宿主体内的不断发育和给药时间的推迟,攻虫后4天和5天给药Ⅲ组的杯状细胞明显减少或消失。
鸡柔嫩艾美耳球虫主要寄生在鸡盲肠,导致肠道形态结构被破坏,肠黏膜损伤,肌层和浆膜等出现不同程度的肿胀[21⁃22]。完整的肠道形态结构是肠道发挥消化吸收、屏障功能的基础,也是肠道组织抵御疾病的前提条件[23]。鸡感染球虫6~7 d后,病理切片观察到盲肠组织形态遭到严重破坏,盲肠上皮细胞内发现大量球虫卵囊[24]。前期研究结果表明,在感染柔嫩艾美耳球虫不同发育阶段的雏鸡饲料中添加常山散后,在攻虫前1天、攻虫当天(子孢子阶段)和攻虫后第2天(滋养体阶段)进行给药,常山散对球虫生长发育具有显著的抑制能力,抗球虫效果良好。在攻虫后第3、4和5天给药,即裂殖体、裂殖子阶段,常山散对球虫的生长发育抑制能力降低,抗球虫效果较差[25]。球虫发育史包括孢子生殖、裂殖生殖和配子生殖。裂殖生殖期对鸡群危害最为严重,产生的大量裂殖子重新入侵未感染的肠道上皮细胞,临床上表现为鸡排出血便、消化和吸收功能下降,甚至死亡[26]。球虫从子孢子入侵到形成卵囊,整个发育史为7 d。一般攻虫后3~4 d,球虫进入裂殖生殖阶段,肠道受到严重破坏,鸡体消化和吸收功能下降,肠内有害物质进入血液,鸡体处于危重状态,此时常山散给药治疗效果较差。
肠道组织作为机体重要的器官,其上皮结构为病原体、毒素和过敏原从管腔向黏膜组织扩散提供了屏障[27⁃29]。肠道屏障主要包括机械屏障、免疫屏障、生物屏障和黏膜屏障。位于上皮细胞外膜顶端的多个蛋白组成的紧密连接是构成肠道机械屏障的重要结构,在肠道屏障功能和肠通透性调节中起关键作用[30]。紧密连接是一种多蛋白复合物,通过调节肠道内的蛋白质来调节肠道通透性。球虫感染破坏肠道紧密连接,降低紧密连接蛋白Occludin和Claudin-1表达,损害肠机械屏障功能[4,31]。本试验结果表明,与Ⅰ组相比攻虫前1天给药至攻虫后第5天给药Ⅱ组和Ⅲ组的Occludin和Claudin-1蛋白表达显著降低(P<0.05);在球虫发育前期,即攻虫前1天、攻虫当天和攻虫后第2天进行给药,与Ⅱ组相比,Ⅲ组的Occludin和Claudin-1的蛋白表达显著提升(P<0.05),但随着球虫在体内的不断发育,在球虫发育的后期,即攻虫后第3、4和5天给药,此时由于球虫对肠道的损伤已经较为严重,故药物对球虫的作用效果较差,蛋白表达明显降低。
在哺乳动物中,盲肠在消化中的作用可以忽略不计;然而在禽类中,盲肠是饲料发酵的重要场所,和禽类健康生长密切相关,盲肠组织的形态和结构可反映禽类消化和吸收等生理功能。杯状细胞分布于肠上皮细胞中,来源于肠隐窝的多能干细胞,参与促进肠上皮细胞的发育和吸收。同时杯状细胞是遍布整个黏膜上皮特化的分泌细胞,通过分泌蛋白质、黏蛋白等多种因子在维持组织稳态方面发挥重要作用。黏蛋白是一种高分子量的糖蛋白,迄今为止,已鉴定出20多个黏蛋白基因。杯状细胞分泌的糖蛋白可以冲洗外来致病因子,防止有害物质接触肠上皮细胞。本试验研究中,柔嫩艾美尔球虫繁殖破坏了盲肠的结构形态,导致大量的杯状细胞丢失和黏蛋白分泌减少,这些现象影响肠黏膜的正常免疫反应。结果表明,与Ⅱ组相比攻虫前1天给药至攻虫后第3天给药Ⅲ组的杯状细胞数和黏蛋白表达显著升高(P<0.05),但随着球虫的不断发育和给药时间的推迟,攻虫后第4天和第5天给药Ⅲ组杯状细胞数和黏蛋白表达明显减少或消失。另外,常山散能够提高感染球虫雏鸡血液中IgG、IgM和IgA的抗体水平,提高机体的体液免疫应答,促进鸡免疫器官的生长发育,增强感染雏鸡的免疫力,从而抑制球虫的发育繁殖,提高抗球虫效果[25]。综上所述,常山散可减轻球虫对宿主肠道的破坏,改善肠道的消化吸收功能,促进肠道屏障蛋白分泌,提高鸡体免疫力,从而发挥抗球虫作用。
本研究结果表明,对感染柔嫩艾美耳球虫不同发育阶段的雏鸡在饲料中添加常山散后,在攻虫前1天、攻虫当天和攻虫后第2天进行给药,常山散可促进球虫感染雏鸡肠道组织损伤的修复,保护肠道组织结构的完整性,减少机械屏障紧密连接蛋白、黏蛋白和肠道杯状细胞的损失,具有良好的抗球虫效果。在攻虫后第3、4和5天给药,此时球虫处于后期发育繁殖阶段,肠组织结构遭到严重破坏,肠紧密连接蛋白表达显著降低,杯状细胞和黏蛋白基本消失,常山散对肠道组织损伤修复作用减弱,对球虫感染雏鸡的治疗效果较差。
  • “十四五”国家重点研发计划项目(2023YFD1800803-4)
  • 国家自然科学基金地区项目(82160730)
  • 石家庄市政府与中国农科院战略合作项目(25010)
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doi: 10.11843/j.issn.0366-6964.2026.01.046
  • 接收时间:2025-04-08
  • 首发时间:2026-09-16
  • 出版时间:2026-01-23
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  • 收稿日期:2025-04-08
基金
“十四五”国家重点研发计划项目(2023YFD1800803-4)
国家自然科学基金地区项目(82160730)
石家庄市政府与中国农科院战略合作项目(25010)
作者信息
    1.甘肃中医药大学药学院,兰州 730000
    2.中国农业科学院兰州畜牧与兽药研究所,农业农村部兽用药物创制 重点实验室,甘肃省中兽医技术创新中心,兰州 730050

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李成义,主要从事中药鉴定与品质评价研究,E-mail:
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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