Ca^(2+)、Mg^(2+)对Caco-2肠道细胞屏障损伤模型的修复作用

REPAIR EFFECT OF Ca^(2+) AND Mg^(2+) ON A CACO-2 INTESTINAL CELL BARRIER INJURY MODEL

目的 建立Caco-2肠道细胞屏障损伤模型,并研究钙、镁对其的保护作用。方法 建立无钙环境诱导的Caco-2肠细胞屏障损伤模型。CCK-8法筛选钙、镁浓度;钙、镁单独或联合暴露后,采用跨膜电阻实验检测细胞单层电阻值;细胞通透性实验检测荧光素的通透性;免疫荧光法测定各组ZO-1、Occludin、Claudin-1蛋白形态;蛋白芯片法测定IL-1α、IL-6、TNF-α表达水平。结果 无钙环境可以构建Caco-2肠细胞屏障损伤模型。与模型组相比,钙组、钙-低镁组和钙-高镁组都能够显著增加跨膜电阻(TEER)值(P<0.05),显著降低荧光黄通透量(P<0.05),并恢复细胞紧密连接蛋白ZO-1、Occludin、Claudin-1的蛋白形态。钙-高镁组下调炎性因子的效果相较更好。结论 钙-镁能够对Caco-2肠细胞屏障损伤起到一定程度的保护作用。[营养学报,2022,44(5):484-490]

Objective To establish Caco-2 intestinal cell barrier injury model and study the protective effect of calcium and magnesium. Methods The model of Caco-2 intestinal cell barrier injury induced by calcium-free environment was established. Calcium and magnesium concentrations were screened by CCK-8 method. Monolayer resistance was measured by transmembrane resistance test after calcium and magnesium exposure alone or in combination. The permeability of fluorescein was detected by cell permeability assay. The distribution of ZO-1, Occludin and Claudin-1 was determined by immunofluorescence method. The expression levels of IL-1α, IL-6 and TNF-α were determined by protein chip assay. Results Caco-2 intestinal cell barrier injury model was established successfully in a calcium-free environment. Compared with the model group, calcium group, calcium-low magnesium group and calcium-high magnesium group could significantly increase the value of transmembrane resistance(TEER)(P<0.05), decreased the permeability of fluorescent yellow(P<0.05) and restored the distribution of tight junction proteins ZO-1, Occludin and Claudin-1. The calcium-high magnesium group was better in down-regulating inflammatory factors.Conclusion Calcium-magnesium can protect Caco-2 intestinal cell barrier to a certain extent. [ACTA NUTRIMENTA SINICA, 2022,44(5):484-490]