猪肠道DMT1及FP1基因转录后调控及其对铁吸收的影响及机制研究

Iron is one of the essential trace elements for animal growth and development. Nowadays, iron deficiency in piglets and growing pigs are particularly serious in the modern pig production. Because the bioavailability of iron additives are lower in the intestine, so it is a good way to resolve the pig iron deficiency by improving the efficiency of iron absorption using effective expression of genes involved in iron absorption in the intestine. In this study, first, it is widely used to predict candidate microRNAs regulating genes involving in absorption and transport of ferrous iron (DMT1 and FP1) in intestine by bioinformatics software (Targetscan and miRDB) and HeLa cells line transfected with microRNA over-expression vector and Dual-Luciferase reporter assay system;Second,function of candidate microRNAs are identified using Caco-2 cells line transfected with microRNA over-expression vector or anti-miRNA oligonucleotide; Third, function of candidate microRNAs are identified again using piglet of iron deficiency injected with adenovirus vector or anti-miRNA oligonucleotide via the superior vena cava. This study will identify the microRNAs targeted regulation of the intestinal divalent iron transporter (DMT1 and FP1),then provides a theoretical basis in order to directional regulation of intestinal ferrous iron absorption.

铁是猪生长发育必需的微量元素之一，集约化养猪生产中，仔猪及生长猪极易缺铁。由于饲料中外源添加铁制剂的吸收效率极低，因此内源性增强猪肠道铁吸收转运蛋白表达，提高铁吸收转运效率，是改善猪铁缺乏的有效途径。本研究采用生物信息学软件Targetscan及miRDB预测并结合microRNA过表达载体及荧光素酶报告基因载体转染HeLa细胞方法筛选获得靶向调控猪肠道二价铁吸收转运关键基因DMT1及FP1的候选microRNAs；通过离体Caco-2细胞模型转染候选microRNA过表达载体候选及microRNA反义单链结合缺铁仔猪前腔静脉注射候选microRNA腺病毒载体及候选microRNA反义单链anti-miR方式对靶向调控猪肠道二价铁吸收转运关键基因DMT1或FP1的microRNAs进行功能验证。该研究可明确靶向调控DMT1及FP1的microRNAs，为定向调控肠道二价铁吸收提供理论依据。