Figure 5.

Chlorogenic acid (CGA) attenuates tumor necrosis factor-α (TNF-α) induced inflammation and injury in IPEC-J2 cells. (A) Analysis of cell apoptosis by flow cytometric assays. In each diagram, Q1 represents the percentage of nonviable and necrotic cells, Q2 represents the percentage of late apoptotic cells, Q3 represents the percentages of early apoptotic cells, and Q4 represents the percentage of live cells. IPEC-J2 cells were plated in 24-well plates at a density of 1 × 10⁵ cells/well and pretreated by CGA (50 µM) for 6 h, and the cells were then challenged by TNF-α (50 ng/mL) for 3 h (n = 6). (B) Assays of cell viability. (C) qPCR analysis of the expressions of claudin-1. (D) Expressions of critical inflammation-related molecules. (E) Expressions of critical apoptosis-related proteins. (F) Western-blot analysis of critical signaling proteins involved in the NF-kappa-B inhibitor alpha (IκBα)/ nuclear factor-κB (NF-κB) pathway. CON, cells without being treated; TNF, cells were only treated by 50 ng/mL TNF-α; TCGA, cells were pretreated by 50 µM CGA for 6 h and were then treated by 50 ng/mL TNF-α for 3 h; CGA, cells were only treated by 50 µM CGA. * p < 0.05, ** p < 0.01.