为探讨丁酸钠对肠道寡肽转运载体(PepT1)及钠氢交换载体(NHE2、NHE3)mRNA表达丰度的调控,取刚出生未吮乳的仔猪小肠黏膜组织,建立猪小肠上皮细胞(IEC)分离及原代培养方法。分别用0,2,4,8 mmol/L的丁酸钠处理体外培养的猪小肠黏膜上皮细胞96 h后,提取细胞总RNA,以18S rRNA为内标基因,用实时荧光定量RT-PCR法(SYBR GreenⅠ试剂盒)检测PepT1、NHE2及NHE3 mRNA在不同浓度丁酸钠处理细胞中的表达丰度。结果表明,体外培养的猪小肠黏膜上皮细胞用丁酸钠处理96 h后,PepT1和NHE2 mRNA的表达丰度均显著增加(P〈0.05),且呈现剂量依赖效应;NHE3 mRNA表达丰度的剂量效应不明显,只有在高浓度丁酸钠(8 mmol/L)组时才显著高于对照组(P〈0.05)。
In order to research the effects of sodium butyrate on the expression of PepT1,NHE2 and NHE3 mRNA abundances in primary cultured pig intestinal epithelial cell(IEC) in vitro,newborn pig was used to establish the method for isolation and primary culture of IEC.Four levels of sodium butyrate were added to the media for 96 h,respectively.The target genes mRNA abundances were determined by real-time RT-PCR using SYBR Green I RTPCR mix Kit.Cells were collected for analyzing the PepT1 and NHE2 mRNA abundances by real-time RT-PCR method.The results showed that the expression of PepT1 and NHE2 mRNA abundances was increased in sodium butyrate treatment and exhibiting dosage-depended also.The dosage-depended effect of NHE3 mRNA in sodium butyrate treatment was not significant.