Lee So-Rok, Hwang Hye-Jeong, Yoon Ju-Gyeong, Bae Eun-Young, Goo Kyo-Suk, Cho Sang-Joon, Cho Jin Ah
Department of Food and Nutrition, Chungnam National University, 99, Daehak-ro, Yuseong-gu, Daejeon 34134, Korea.
Department of Agrofood Resources, National Institute of Agricultural Sciences, RDA, Wanju, Jeonbuk 55365, Korea.
Nutr Res Pract. 2019 Apr;13(2):95-104. doi: 10.4162/nrp.2019.13.2.95. Epub 2019 Mar 11.
BACKGROUND/OBJECTIVES: Inflammatory Bowel Disease (IBD) has rapidly escalated in Asia (including Korea) due to increasing westernized diet patterns subsequent to industrialization. Factors associated with endoplasmic reticulum (ER) stress are demonstrated to be one of the major causes of IBD. This study was conducted to investigate the effect of () on ER stress.
MATERIALS/METHODS: Mouse embryonic fibroblast (MEF) cell line and polarized Caco-2 human intestinal epithelial cells were treated with crude extract of the fruit (LF). Paracellular permeability was measured to examine the effect of tight junction (TJ) integrity. The regulatory pathways of ER stress were evaluated in MEF knockout (KO) cell lines by qPCR for interleukin (IL) 6, IL8 and XBP1 spliced form (XBP1s). Immunoglobulin binding protein (BiP), XBP1s and CCAAT/enhancer-binding homologous protein (CHOP) expressions were measured by RT-PCR. Scanning Ion Conductance Microscopy (SICM) at high resolution was applied to observe morphological changes after treatments.
Exposure to LF extract strengthened the TJ, both in the presence and absence of inflammation. In polarized Caco-2 pretreated with LF, induction in the expression of proinflammatory marker IL8 was not significant, whereas ER stress marker XBP1s expression was significantly increased. In wild type (wt) MEF cells, IL6, CHOP and XBP1 spliced form were dose-dependently induced when exposed to 12.5-50 µg/mL extract. However, absence of XBP1 or IRE1α in MEF cells abolished this effect.
Results of this study show that LF treatment enhances the barrier function and reduces inflammation and ER stress in an IRE1α-XBP1-dependent manner. These results suggest the preventive effect of LF on healthy intestine, and the possibility of reducing the degree of inflammatory symptoms in IBD patients.
背景/目的:由于工业化后西方化饮食模式的增加,炎症性肠病(IBD)在亚洲(包括韩国)迅速增加。内质网(ER)应激相关因素被证明是IBD的主要原因之一。本研究旨在探讨()对ER应激的影响。
材料/方法:用该果实(LF)的粗提物处理小鼠胚胎成纤维细胞(MEF)系和极化的Caco-2人肠上皮细胞。测量跨细胞通透性以检查紧密连接(TJ)完整性的影响。通过qPCR检测白细胞介素(IL)6、IL8和XBP1剪接形式(XBP1s),评估MEF基因敲除(KO)细胞系中ER应激的调节途径。通过RT-PCR测量免疫球蛋白结合蛋白(BiP)、XBP1s和CCAAT/增强子结合同源蛋白(CHOP)的表达。应用高分辨率扫描离子电导显微镜(SICM)观察处理后的形态变化。
暴露于LF提取物可增强TJ,无论有无炎症。在用LF预处理的极化Caco-2中,促炎标志物IL8的表达诱导不显著,而ER应激标志物XBP1s的表达显著增加。在野生型(wt)MEF细胞中,当暴露于12.5-50μg/mL提取物时,IL6、CHOP和XBP1剪接形式呈剂量依赖性诱导。然而,MEF细胞中XBP1或IRE1α的缺失消除了这种作用。
本研究结果表明,LF处理以IRE1α-XBP1依赖性方式增强屏障功能并减轻炎症和ER应激。这些结果提示LF对健康肠道的预防作用,以及降低IBD患者炎症症状程度的可能性。