Protective Effect of Total Polyphenolic Extracts from Pomelo Levels on LPS-induced Intercellular Hyperpermeability in Caco-2 Cells
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Abstract:
To investigate the protective effect of total polyphenolic extracts from pomelo levels (PLTPE) on lipopolysaccharide (LPS, 2 g/mL) induced hypepermeability in Caco-2 cells in this paper. The cell survival rate was measured by MTT assay. The cellular level of lactate dehydrogenase (LDH) was determined by kit. Enzyme linked immunosorbent assay (ELISA) was used to determine the level of Interlukin (IL)-1β, IL-8, and tumor necrosis factor (TNF)-α. The transepithelial cell resistance (TEER) and fluorescein dextran (FD40) permeability were used to evaluate the permeability of cells. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of IL-1β, IL-8, TNF-α, occludin, claudin-1, ZO-1, and myosin light chain kinase (MLCK). PLTPE treatment significantly increased the cell survival rate and inhibited the spillover of LDH in Caco-2 cells. PLTPE treatment effectively inhibited the secretion of inflammatory cytokines (IL-1β, IL-8, TNF-α) and reduced their mRNA transcription in Caco-2 cells. At the same time, PLTPE also significantly increased the mRNA levels of occludin, claudin-1, ZO-1, and inhibit the MLCK mRNA transcription to improve the intercellular permeability in LPS treated Caco-2 cells. These results suggest that the PLTPE showed a strong anti-inflammatory activity, and can improve the LPS induced high permeability of Caco-2 cells may associate with regulating the mRNA transcription of tight junction related factors.
