Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István u. 2, Budapest, 1078, Hungary,
Inflammation. 2014 Feb;37(1):247-60. doi: 10.1007/s10753-013-9735-7.
A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 epithelial cells as upper layer, was developed to study the effects of lipopolysaccharides (LPS) on the gene expression profile of pro-inflammatory cytokines (interleukin-8 (IL-8) and tumor necrosis factor-α) and CYP enzymes (CYP1A1, CYP1A2, CYP3A29). The barrier integrity of IPEC-J2 cells was investigated by transepithelial electrical resistance measurements and by fluorescein isothiocyanate-dextran-based test. Basolateral IL-8 production was significantly elevated in LPS-treated IPEC-J2 and primary hepatocyte mono-cultures as well as in the co-culture system, in a dose-independent manner. The LPS-induced changes in the expression of the CYP1A2 and CYP3A29 genes in hepatocyte mono-cultures differed from those in co-culture after LPS treatment on the apical side of the IPEC-J2 cell layer. CYP1A2 was downregulated by the LPS treatment in mono-cultures but upregulated at 10 μg/ml LPS in co-culture; gene expression of CYP3A29 showed no significant LPS-induced change in the hepatocyte mono-culture but was significantly downregulated in co-culture. The newly established co-culture system capable of mimicking enterohepatic interplay in LPS-induced inflammatory responses in vitro can be used in the future for reliable screening of potential anti-inflammatory compounds.
建立了一个猪肠肝共培养体系，以下皮细胞为底层，IPEC-J2 上皮细胞为上层，用于研究脂多糖（LPS）对促炎细胞因子（白细胞介素-8（IL-8）和肿瘤坏死因子-α）和 CYP 酶（CYP1A1、CYP1A2、CYP3A29）基因表达谱的影响。通过跨上皮电阻测量和荧光素异硫氰酸酯-葡聚糖基础试验来研究 IPEC-J2 细胞的屏障完整性。LPS 处理的 IPEC-J2 和原代肝细胞单培养以及共培养系统中，底侧 IL-8 的产生均显著增加，且呈剂量非依赖性。LPS 处理后，在 IPEC-J2 细胞层的顶侧，LPS 对 CYP1A2 和 CYP3A29 基因表达的影响在肝细胞单培养和共培养中存在差异。在单核培养中，CYP1A2 被 LPS 下调，但在共培养中，在 10μg/ml LPS 时被上调；在单核培养中，CYP3A29 的基因表达没有明显的 LPS 诱导变化，但在共培养中明显下调。该新建立的共培养系统能够模拟 LPS 诱导的炎症反应中的肠肝相互作用，将来可用于可靠筛选潜在的抗炎化合物。
