Comparative evaluation of the modulatory role of 1,25-dihydroxy-vitamin D_(3) on endoplasmic reticulum stress-induced effects in 2D and 3D cultures of the intestinal porcine epithelial cell line IPEC-J2  

作　　者：Gaiping Wen Klaus Eder Robert Ringseis 

机构地区：[1]Institute of Animal Nutrition and Nutrition Physiology,Justus Liebig University Giessen,Heinrich‑Buff‑Ring 26‑32,Giessen 35392,Germany [2]Center for Sustainable Food Systems,Justus Liebig University Giessen,Senkenbergstrase 3,Giessen 35390,Germany

出　　处：《Journal of Animal Science and Biotechnology》2025年第2期741-757,共17页畜牧与生物技术杂志(英文版)

摘　　要：Background The use of conventional two-dimensional(2D)culture of the porcine intestinal epithelial cell(IEC)line IPEC-J2 in animal nutrition research has the disadvantage that IEC function is studied under unphysiological conditions,which limits the ability of transferring knowledge to the in vivo-situation.Thus,the aim of the present study was to establish a more convincing and meaningful three-dimensional(3D)culture of IPEC-J2 cells,which allows to study cell function in a more tissue-like environment,and to compare the effect of the endoplasmic reticulum(ER)stress inducer tunicamycin(TM)on ER stress indicators and the expression of tight junction proteins(TJP),inflammatory and apoptosis-related genes and the modulatory role of 1,25-dihydroxy-vitamin D_(3)(1,25D_(3))on these parameters in 2D and 3D cultures of IPEC-J2 cells.Results A published protocol for 3D culture of Caco-2 cells was successfully adopted to IPEC-J2 cells as evident from fully differentiated 3D IPEC-J2 spheroids showing the characteristic spherical architecture with a single layer of IPEC-J2 cells surrounding a central lumen.Treatment of 2D IPEC-J2 cells and 3D IPEC-J2 spheroids with TM for 24 h markedly increased m RNA and/or protein levels of the ER stress target genes,heat shock protein family A(Hsp70)member 5(HSPA5)and DNA damage inducible transcript 3(DDIT3),whereas co-treatment with TM and 1,25D_(3) did not mitigate TM-induced ER stress in IPEC-J2 cells in the 2D and the 3D cell culture.In contrast,TM-induced expression of pro-inflammatory[interleukin-6(IL6),IL8]and pro-apoptotic genes[BCL2 associated X,apoptosis regulator(BAX),caspase 3(CASP3),CASP8]and genes encoding TJP[TJP1,claudin 1(CLDN1),CLDN3,occludin(OCLN),cadherin 1(CDH1),junctional adhesion molecule 1(JAM1)]was reduced by co-treatment with TM and 1,25D_(3) in 3D IPEC-J2 spheroids but not in the 2D cell culture.Conclusions The effect of 1,25D_(3) in the IPEC-J2 cell culture is dependent on the culture model applied.While 1,25D_(3) does not inhibit TM-induced expression of gene

关 键 词：1 25-Dihydroxy-vitaminD_(3) 3D cell culture Endoplasmic reticulum stress Intestinal barrier PIG 

分 类 号：S816[农业科学—饲料科学] S828[农业科学—畜牧学]