海洋天然产物壳寡糖抑制细菌生长和黏附的作用评价  

Evaluation of Inhibitory Effect of Marine Natural Product Chitosan Oligosaccharide on Bacterial Growth and Adhesion

作　　者：温佳颖 牛雪婷 陈圣威 陈宗周 王新晨 吴淑婷 巨向红[1,2] 刘晓曦 雍艳红 WEN Jiaying;NIU Xueting;CHEN Shengwei;CHEN Zongzhou;WANG Xinchen;WU Shuting;JU Xianghong;LIU Xiaoxi;YONG Yanhong(Department of Veterinary Medicine,College of Coastal Agricultural Sciences,Guangdong Ocean University,Zhanjiang 524000,China;Shenzhen Institute of Guangdong Ocean University,Shenzhen 518108,China)

机构地区：[1]广东海洋大学滨海农业学院动物医学系,湛江524000 [2]广东海洋大学深圳研究院,深圳518108

出　　处：《动物营养学报》2022年第5期3297-3306,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金(31472243,31902314);广东省自然科学基金(2019A1515011142)。

摘　　要：海产品加工副产物壳寡糖(COS)具有抗菌、消炎和黏膜修复等重要生物活性。为了解COS作为替抗天然产物的功能和作用,本研究检测了COS对致病性大肠杆菌、肠炎沙门菌、金黄色葡萄球菌和福氏志贺菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC),通过羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记观察了COS对4种致病菌在猪肠道上皮细胞IPEC-J2黏附的抑制率,通过qPCR检测了细菌黏附相关基因的表达水平。结果表明:COS对这4种肠道致病菌均有良好的抑菌作用,其中对致病性大肠杆菌抑杀效果最明显;COS可抑制4种致病菌对IPEC-J2细胞的黏附作用,以对致病性大肠杆菌的黏附抑制效果最好,黏附率能下降至50%以下;COS对致病菌侵染的IPEC-J2细胞间黏附分子-1(ICAM-1)基因相对表达量的上调有极显著抑制作用(P<0.01),对黏蛋白(MUC)2、MUC4及闭锁蛋白(Occludin)基因相对表达量有显著上调作用(P<0.05)。结果说明,COS有明显的抑菌、黏附抑制和黏膜保护作用,具有作为替抗减抗功能饲料添加剂开发的潜力。Chitosan oligosaccharide(COS),a by-product of marine processing,had important biological activities such as antibacterial,anti-inflammatory and mucosal repair. In order to understand the functions of COS as a natural product of alternative antibiotic,the minimal inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)of COS to enteropathogenic Escherichia coli,Salmonella enteritis,Staphylococcus aureus and Shigella were detected. The adhesion rate of COS on pathogenic bacteria in porcine intestinal epithelial cells IPEC-J2 were detected by 5,6-carboxyfluorescein diacetate succinimidy ester(CFSE). The expression level of adhesion related genes was detected by qPCR. The results showed that COS had good bacteriostasis effect on these four intestinal pathogenic bacteria,especially on enteropathogenic Escherichia coli;and COS could inhibit the adhesion of four pathogenic bacteria to IPEC-J2 cells,especially on enteropathogenic Escherichia coli,and the adhesion rate could be reduced to less than 50%. COS significantly inhibited the upregulation of adhesion molecule-1(ICAM-1)in IPEC-J2 cells infected by pathogenic bacteria(P<0.01),and significantly up-regulated mucin(MUC)2,MUC4 and Occludin(P<0.05). The results show that COS has obvious effects of bacteriostasis,adhesion inhibition and mucosal protection. It has the function of developing substitute anti-reducing anti-feed additive.[Chinese Journal of Animal Nutrition,2022,34(5):3297-3306]

关 键 词：壳寡糖 细菌黏附 肠上皮 肠道致病菌 

分 类 号：S816[农业科学—饲料科学]