摘要
以仔猪肠上皮细胞(IPEC-J2)为研究对象,用大豆7S球蛋白作为诱导因素,通过检测细胞活力、细胞跨膜电阻(TEER)、荧光素钠渗透率、细胞膜完整性相关指标与紧密连接蛋白的mRNA表达情况,探究维生素A(V_(A))对大豆7S球蛋白致IPEC-J2细胞屏障功能损伤的影响。结果显示:5.0 mg·mL^(-1)大豆7S球蛋白导致IPEC-J2活力、TEER与ZO-1、Claudin-1、Occludin的mRNA表达量极显著(P<0.01)降低,而荧光素钠渗透率和细胞培养液中碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)、二胺氧化酶(DAO)、肠型脂肪酸结合蛋白1(IFABP1)的含量极显著(P<0.01)升高;添加0.1和1μmol·L^(-1)V_(A)极显著提高了细胞活力(P<0.01),0.1~10μmol·L^(-1)V_(A)显著(P<0.05)缓解了大豆7S球蛋白导致的TEER、荧光素钠渗透率、细胞膜完整性相关指标与紧密连接蛋白mRNA表达的变化;10000μmol·L^(-1)V_(A)反而加剧了这些影响。总体上,0.1~10μmol·L^(-1)V_(A)能通过保护细胞活力和细胞完整性、影响紧密连接蛋白的mRNA表达对大豆7S球蛋白导致的IPEC-J2细胞屏障功能损伤发挥保护作用,而10000μmol·L^(-1)V_(A)反而加重了这种损伤。
To explore effect of vitamin A(V_(A))on barrier function damage of piglet intestinal epithelial cells(IPEC-J2)induced by soybean 7S globulin,took IPEC-J2 as the research object and soybean 7S globulin as the inducing factor,cell viability,trans-epithelial electrical resistance(TEER),permeability of sodium fluorescein,cell membrane integrity related indicators and tight junction protein mRNA expression were assessed.The results showed that 5 mg·mL^(-1) soybean 7S globulin caused IPEC-J2 viability,TEER,ZO-1,Claudin-1 and Occludin mRNA expression significantly reduced(P<0.01),and permeability of fluorescein sodium,content of alkaline phosphatase(ALP),lactate dehydrogenase(LDH),diamine oxidase(DAO)and intestinal fatty acid binding protein 1(IFABP1)in the cell culture fluid significantly increased(P<0.01).0.1 and 1μmol·L^(-1)V_(A) could significantly protect cell viability(P<0.01),0.1-10μmol·L^(-1)V_(A) could significantly(P<0.05)alleviate the changes of TEER,fluorescein sodium permeability,cell membrane integrity related indicators and tight junction protein mRNA expression caused by 7S globulin;10000μmol·L^(-1)V_(A) exacerbated these effects.The results indicated that 0.1-10μmol·L^(-1)V_(A) could protect the cell barrier function of IPEC-J2 caused by soybean 7S globulin through protecting cell viability and cell integrity,and affecting expression of tight junction protein mRNA.However,10000μmol·L^(-1)V_(A) aggravated this damage.
作者
杨菊
邓俊良
夏江英
宋天浩
庞莲凤
任志华
YANG Ju;DENG Junliang;XIA Jiangying;SONG Tianhao;PANG Lianfeng;REN Zhihua(College of Veterinary Medicine/Key Laboratory of Animal Disease and Human Health of Sichuan Province/Key Laboratory of Environmental Hazards and Animal Diseases of Sichuan Province Colleges and Universities, Sichuan Agriculture University, Chengdu 611130, China)
出处
《浙江农业学报》
CSCD
北大核心
2021年第11期2026-2033,共8页
Acta Agriculturae Zhejiangensis
基金
国家“十三五”重点研发计划(2016YFD0501204)。
