摘要
本研究通过测定大豆抗原β-Conglycinin酶解肽(52ku)对仔猪空肠上皮细胞(IPEC)通透性及紧密连接蛋白Occludin与ZO-1基因mRNA表达的影响,为探索其对肠道损伤的相关机制提供依据。本试验分别采用不同浓度(0、0.125、0.25、0.5、1、2mg·mL-1)52ku酶解肽处理IPEC 2、4、6、8、12、24h后,通过测定细胞存活力(MTT值)、跨上皮细胞电阻值(TEER)以及细胞培养液中碱性磷酸酶(AP)的活性,检测52ku酶解肽对细胞膜通透性的影响。并选取0.5mg·mL-1 52ku酶解肽处理IPEC 24h后,用实时荧光定量PCR方法检测Occludin与ZO-1基因mRNA表达量的变化情况。结果表明,52ku酶解肽处理仔猪空肠上皮细胞后,MTT值和TEER呈时间-剂量依赖性降低(P<0.05)。24h处理后,AP活性呈显著升高趋势(P<0.05)。与对照组相比,52ku酶解肽可使Occludin和ZO-1基因mRNA相对表达量均呈显著下降趋势(P<0.05)。总之,52ku酶解肽使仔猪空肠上皮细胞通透性增加,并降低Occludin或ZO-1的mRNA表达。
The objective of this study was to evaluate the effects ofβ-Conglycinin hydrolyzed peptide(52ku)on the permeability and expression of tight junction protein Occludinand ZO-1in piglet intestinal epithelial cells(IPEC),which would provide a basis for exploring the mechanisms of intestinal damage.IPEC were treated with different concentrations(0,0.125,0.25,0.5,1and 2mg·mL-1)hydrolyzed peptide for 2,4,6,8,12,24 h.The effects of hydrolyzed peptide(52ku)on cell membrane permeability were determined by measuring the cell viability(MTT values),transepithelial resistance values(TEER)and alkaline phosphatase(AP)activity in cell culture medium.The mRNA expression of Occludin and ZO-1were detected by real-time PCR after IPEC treated with 0.5 mg·mL-1 hydrolyzed peptide for 24 h.The results showed that MTT and TEER of intestinal epithelial cells treated with hydrolyzed peptide(52ku)with 0.5 mg·mL-1were decreased by a time-dose dependent manner(P<0.05).With the increase concentration of hydrolyzed peptide(52ku),AP activity in 24 htreatment group had a significant increased trend(P<0.05)by dose dependent manner.Also,hydrolyzed peptide(52ku)could decrease the relative expression of Occludinand ZO-1mRNA in IPEC(P<0.05).In conclusion,hydrolyzed peptide(52ku)destructed the permeability and reduced Occludinor ZO-1mRNA expression of tight junction protein in IPEC.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2015年第5期768-773,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金青年资金项目(31101719)
省科技发展计划项目青年科研基金(201201098)
吉林省教育厅"十二五"科学技术研究规划项目
