摘要
【目的】分析雏鸡感染禽致病性大肠埃希菌(Avian pathogenic Escherichia coli,APEC)及其PhoP/Q缺失株后β-防御素6(AvBD6)在小肠内的分布和表达规律。【方法】将14日龄雏鸡随机分为对照组、APEC攻毒组和PhoP/Q缺失株攻毒组,其中对照组雏鸡腿肌注射生理盐水0.5mL/只,APEC和PhoP/Q缺失株攻毒组雏鸡腿肌分别接种相同剂量1×106 CFU/mL的菌液,分别于攻毒后0,6,12,24,36和48h采集各组鸡小肠样品,用荧光定量PCR(FQPCR)方法对各组织内AvBD6mRNA的表达量进行检测,同时采用免疫组化(IHC)技术对AvBD6蛋白分布进行观察。【结果】FQ-PCR结果显示,感染后24~48h,APEC攻毒组雏鸡十二指肠、空肠、回肠的AvBD6mRNA表达量显著高于对照组(P<0.05),于48h达到峰值且显著高于0h(P<0.05);在同一时间下,PhoP/Q缺失株攻毒组AvBD6mRNA的表达量显著高于APEC攻毒组(P<0.05)。IHC结果显示,AvBD6主要分布于各小肠段的绒毛上皮细胞和肠腺部分。【结论】APEC能够诱导雏鸡肠道内AvBD6的表达,且PhoP/Q可能参与APEC调控AvBD6的抗感染作用。
【Objective】This study investigated the distribution and expression pattern of AvBD6 in small intestine of chicks infected with original and PhoP/Q mutant strain of avian pathogenic Escherichia coli（APEC）.【Method】The chicks at the age of 14 days were randomly divided into the blank control group,the APEC infection group and the PhoP/Q mutant infection group.The experimental group was inoculated with 0.5mL of the APEC and its PhoP/Q deletion strain suspension containing 1×10^6 CFU/mL through leg,while the control group was given 0.5mL of PBS.Small intestine samples（duodenum,jejunum,and ileum）were collected within 48h（0,6,12,24,36 and 48h）after infection.The levels of AvBD6 mRNA at each observation time point were detected with fluorescence quantitative PCR（FQ-PCR）,and the protein distribution was analyzed using immunohistochemistry（IHC）.【Result】The consequence of FQ-PCR showed that the group infected with original strain of APEC had significantly higher AvBD6mR-NA expression than that of the control group within 24-48h（P〈0.05）and it reached the maximum at48h（P〈0.05）,but distinctly lower than the group infected with PhoP/Q mutant strain at the same time（P〈0.05）.The result of IHC showed that AvBD6 mainly distributed in villi epithelial cells and glandular portion of each part of the small intestine.【Conclusion】APEC could induce the expression of AvBD6 in small intestine of chicks and PhoP/Q may be involved in the regulation of anti-infection effects of AvBD6.
作者
李少参
王惠珂
祁克宗
张煜
薛挺
涂健
周秀红
刘红梅
Shaocan WANG Huike QI Kezong ZHANG Yu XUE Ting TU Jian ZHOU Xiuhong LIU Hongmei(College of Animal Science and Technology ,Anhui Agriculture University, He f ei,Anhui 230036, Chin)
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2017年第2期44-48,55,共6页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家自然科学基金项目(31372402)
安徽省自然科学基金项目(1508085SQC206)
