维生素C对β-伴大豆球蛋白诱导的仔猪肠上皮细胞炎性损伤的保护作用  

Protective effect of vitamin C in piglet intestinal epithelial cells inflammatory injury induced byβ-conglycinin

作　　者：夏江英 杨菊 宋天浩 庞莲凤 叶婷 任志华[1] 邓俊良[1] XIA Jiangying;YANG Ju;SONG Tianhao;PANG Lianfeng;YE Ting;REN Zhihua;DENG Junliang(College of Veterinary Medicine, Sichuan Agricultural University,Chengdu 611130, China)

机构地区：[1]四川农业大学动物医学院,四川成都611130

出　　处：《浙江农业学报》2021年第11期2017-2025,共9页Acta Agriculturae Zhejiangensis

基　　金：国家“十三五”重点研发计划(2016YFD0501204)。

摘　　要：为分析不同浓度维生素C(vitamin C,V_(C))对β-伴大豆球蛋白(7S)诱导的仔猪肠上皮细胞(IPEC-J2)炎性损伤的保护作用,取对数生长期的IPEC-J2用于试验,随机分为对照组、7S模型组(5 mg·mL^(-1)7S)和V_(C)(25、50、100、200、400、600、800、1000μmol·L^(-1))保护组。细胞培养24 h后采用CCK-8法检测细胞存活率,倒置显微镜观察细胞形态学变化,ELISA法检测细胞上清液中LDH、ALP、DAO、IFABP1含量和IL-1β、IL-6、TNF-α、IL-4、IL-10的分泌水平,用qRT-PCR法检测IL-1β、IL-6、TNF-α、IL-4和IL-10 mRNA相对表达量。结果显示:7S可显著(P<0.01)降低IPEC-J2活力、破坏细胞膜完整性,上调细胞促炎性因子和下调抗炎因子的产生;与7S模型组相比,同时添加7S和V_(C)的试验组细胞活力增加,细胞上清液中LDH、ALP、DAO、IFABP1含量显著(P<0.01)降低,促炎因子IL-1β、IL-6、TNF-α分泌水平降低,抗炎因子IL-4和IL-10的分泌水平升高。因此,不同浓度V_(C)均可保护由7S诱导引起的仔猪肠上皮细胞损伤,100μmol·L^(-1) V_(C)的修复和保护作用最佳。To investigate protective effects of different concentrations of vitamin C(V_(C))on inflammatory injury of piglet intestinal epithelial cells(IPEC-J2)induced byβ-conglycinin(7S).IPEC-J2 in logarithmic growth phase were used for test.The cells were randomly divided into control group,7S model group(5 mg·mL^(-1)7S),and V_(C)(25,50,100,200,400,600,800,1000μmol·L^(-1))protected groups.After cells were cultured for 24 h,cell viability was assessed by CCK-8 assay and morphological changes of cells were observed by an inverted microscope.Contents of LDH,ALP,DAO,IFABP1,IL-1β,IL-6,TNF-α,IL-4 and IL-10 in the cell supernatant were detected by ELISA assay.The relative expression of IL-1β,IL-6,TNF-α,IL-4 and IL-10 mRNA were detected by qRT-PCR.The results showed that 7S significantly(P<0.01)reduced the viability of IPEC-J2 and destroyed the integrity of cell membranes.7S also elevated pro-inflammatory and decreased anti-inflammatory factors in cells.Compared with 7S model group,cell viability in the test group added with 7S and V_(C) at the same time was significantly increased,and contents of LDH,ALP,DAO,IFABP1 in cell supernatant were significantly(P<0.01)reduced.Compared with 7S model group,V_(C) protection groups reduced the secretion levels of cell pro-inflammatory factors IL-1β,IL-6 and TNF-α,and increased the secretion levels of anti-inflammatory factors IL-4 and IL-10.Therefore,various doses of V_(C) protected against intestinal epithelial cell injury in piglets induced by 7S,and 100μmol·L^(-1) V_(C) had the best repair and protection effect on 7S injured IPEC-J2.

关 键 词：维生素C Β-伴大豆球蛋白 仔猪肠上皮细胞 膜完整性 细胞因子 

分 类 号：S852.4[农业科学—基础兽医学]