Journal: The Kaohsiung Journal of Medical Sciences

Article Title: In Vitro Evaluation of the Inhibitory Effects of Linarin on Histamine‐Induced Expression of Proinflammatory Cytokines, Mucin 5AC , and Aquaporin 5 in Human Nasal and Bronchial Epithelial Cells

doi: 10.1002/kjm2.70114

Figure Lengend Snippet: Roles of NF‐κB and MAPK pathways in regulating inflammatory cytokine secretion in linarin‐treated cells. (A, B) hNECs and BEAS‐2B cells were treated with dexamethasone (100 nM) or the inhibitors of NF‐κB (PDTC, 10 μM), ERK (U0126, 10 μM), JNK (SP600125, 10 μM), and p38 (SB203580, 30 μM) for 24 h. The cytotoxicity effects of dexamethasone and these inhibitors were evaluated by CCK‐8 assay. (C–H) hNECs and BEAS‐2B cells were pretreated with linarin or/and the inhibitors of NF‐κB (PDTC, 10 μM), ERK (U0126, 10 μM), JNK (SP600125, 10 μM), and p38 (SB203580, 30 μM) for 1 h, and then stimulated with histamine (100 μM) for 24 h. The secretion levels of IL‐6, IL‐8, and MCP‐1 in the medium were detected by ELISA. Cells pretreated with dexamethasone (100 nM) for 1 h and then stimulated with histamine (100 μM) for 24 h were used as a positive control. * p < 0.05, ** p < 0.01, and *** p < 0.001 versus relative group. SP, SP600125. SB, SB203580. Dex, dexamethasone.

Article Snippet: The levels of IL‐6, IL‐8, and MCP‐1 were determined using a Human IL‐6 Quantikine ELISA Kit (QK206; R&D Systems, Minneapolis, MN), Human IL‐8/CXCL8 Quantikine ELISA Kit (D8000C; R&D Systems), and Human CCL2/MCP‐1 Quantikine ELISA Kit (DCP00; R&D Systems) following the manufacturer's protocol.

Techniques: CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Positive Control