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Effects of MCL on oxidative stress and inflammatory cytokine expression in BEAS-2B cells co-induced by IL-4 and TNF-α. (A) Cell viability of BEAS-2B cells treated with different concentrations of MCL (1, 5, 10, and 20 μM) for 24 h. (B – E) Protein levels of CCL11 (B) , CCL24 (C) , IL-6 (D) , and IL-8 (E) in the supernatant of BEAS-2B cells across different groups. (F – G) Intracellular ROS levels of BEAS-2B cells across different groups, measured using a DCFH-DA probe by Multi-Mode Microplate Reader and fluorescence microscopy. In panel A, the Control group received DMSO. The MCL1, MCL5, MCL10, and MCL20 groups were treated with 1, 5, 10, and 20 μM of MCL, respectively. In panels B–G, the Control group received DMSO. The IL-4+TNF-α group was treated with 20 ng/mL each of IL-4 and TNF-α. The MCL1, MCL5, MCL10, and MCL20 groups were co-treated with IL-4 and TNF-α plus 1, 5, 10, and 20 μM of MCL, respectively. Data in panels A–E are presented as mean ± standard deviation from 3 independent experiments. Statistical analysis was performed using one-way ANOVA. Compared with the Control group: ## p < 0.01; Compared with the IL-4+TNF-α group: ∗ p < 0.05, ∗∗ p < 0.01.

Journal: The World Allergy Organization Journal

Article Title: Protective effect of micheliolide against inflammation and oxidative stress in asthma through the MAPK/NF-κB signalling pathway ☆

doi: 10.1016/j.waojou.2025.101091

Figure Lengend Snippet: Effects of MCL on oxidative stress and inflammatory cytokine expression in BEAS-2B cells co-induced by IL-4 and TNF-α. (A) Cell viability of BEAS-2B cells treated with different concentrations of MCL (1, 5, 10, and 20 μM) for 24 h. (B – E) Protein levels of CCL11 (B) , CCL24 (C) , IL-6 (D) , and IL-8 (E) in the supernatant of BEAS-2B cells across different groups. (F – G) Intracellular ROS levels of BEAS-2B cells across different groups, measured using a DCFH-DA probe by Multi-Mode Microplate Reader and fluorescence microscopy. In panel A, the Control group received DMSO. The MCL1, MCL5, MCL10, and MCL20 groups were treated with 1, 5, 10, and 20 μM of MCL, respectively. In panels B–G, the Control group received DMSO. The IL-4+TNF-α group was treated with 20 ng/mL each of IL-4 and TNF-α. The MCL1, MCL5, MCL10, and MCL20 groups were co-treated with IL-4 and TNF-α plus 1, 5, 10, and 20 μM of MCL, respectively. Data in panels A–E are presented as mean ± standard deviation from 3 independent experiments. Statistical analysis was performed using one-way ANOVA. Compared with the Control group: ## p < 0.01; Compared with the IL-4+TNF-α group: ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: ELISA kits were employed to determine the levels of OVA-specific IgE, IL-4 (M4000B, R&D Systems, Minneapolis, MN, USA), IL-5 (M5000, R&D Systems), IL-13 (mIC50278-1, mlbio, Shanghai, China), IFN-γ (MIF00, R&D Systems), IL-1β (ml098416, Shanghai Enzyme-linked Biotechnology, Shanghai, China), and IL-18 (CK-E20324, Shanghai Enzyme-linked Biotechnology) in mouse samples (serum or BALF), as well as the levels of IL-1β (ml058059, Shanghai Enzyme-linked Biotechnology), IL-18 (ml058055, Shanghai Enzyme-linked Biotechnology), IL-6 (D6050, R&D Systems), and IL-8 (D8000C, R&D Systems) and chemokines CCL11 (DTX00, R&D Systems) and CCL24 (DCC240B, R&D Systems) in the supernatant of BEAS-2B cells based on the manufacturer's protocols.

Techniques: Expressing, Fluorescence, Microscopy, Control, Standard Deviation