Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos

作     者：LI Ting-ting LU Na SHAO Yu-xin ZHANG Li-yang LU Lin LIU Zong-ping LUO Xu-gang LIAO Xiu-dong LI Ting-ting;LU Na;SHAO Yu-xin;ZHANG Li-yang;LU Lin;LIU Zong-ping;LUO Xu-gang;LIAO Xiu-dong

作者机构：Poultry Mineral Nutrition LaboratoryCollege of Animal Science and TechnologyYangzhou UniversityYangzhou 225009P.R.China Mineral Nutrition Research DivisionInstitute of Animal SciencesChinese Academy of Agricultural SciencesBeijing 100193P.R.China College of Veterinary MedicineYangzhou UniversityYangzhou 225009P.R.China 

出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)

年 卷 期：2022年第21卷第7期

页      面：2076-2085页

核心收录：

学科分类：0905[农学-畜牧学] 09[农学] 0901[农学-作物学] 

基　　金：supported by the Key Program of the National Natural Science Foundation of China(31630073) the National Natural Science Foundation of China(31472116) the National Key R&D Program of China(2017YFD0502200) the China Agriculture Research System of MOF and MARA(CARS-41) the Agricultural Science and Technology Innovation Program(ASTIP-IAS09)。 

主　　题：broiler phosphorus transporter phosphorus absorption primary cultured duodenal epithelial cell 

摘      要：The aim of the study was to investigate whether phosphorus(P) transporters, type IIb sodium-dependent phosphate cotransporter(NaP-IIb) and inorganic phosphate transporter 2(PiT2), were directly involved in P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos. The siRNAs against NaP-IIb or PiT2 were designed, synthesized and transfected into primary cultured duodenal epithelial cells of chick embryos. Then, the inhibitory efficiency of siRNAs against NaP-IIb or PiT2 was analyzed, and the most efficacious siRNAs were selected to be used for subsequent P absorption experiments. Briefly, primary cultured duodenal epithelial cells of chick embryos were transfected with either NaP-IIb or PiT2 siRNAs and grown in confluent monolayers on transwell plates. The untransfected or transfected cell monolayers were then incubated in an uptake medium containing 0 or 0.25 mmol L^(–1) of P as KH_(2) PO_(4) to measure the P absorption across duodenal epithelial cell monolayers. The results showed that among the siRNAs designed, si-1372 and si-890 were demonstrated to be the most effective in inhibiting the NaPIIb and PiT2 expressions, respectively. Supplemental P increased(P=0.065) the protein abundance of PiT2 and enhanced(P0.0001) P absorption in primary cultured duodenal epithelial cell of chick embryos. Furthermore, NaPIIb silencing decreased(P=0.07) P absorption across duodenal epithelial cell monolayers, while PiT2 silencing had no effect(P=0.345). It is concluded that the NaP-IIb, but not PiT2, might be directly involved in the P absorption of chick duodenal epithelial cells.