Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Measurement of pro-inflammatory cytokine secretion levels in the co-culture system. Analyses were conducted evaluating ( A ) IL-8 levels in the apical medium (differentiated Caco-2 cells) and ( B ) IL-6 and ( C ) TNF-α levels in the basolateral medium (RAW264.7 cells). Caco-2 cells were pre-treated for 90 min with BD, BFM (1/20, v / v ), and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. Data are expressed as mean ± SD of three experiments conducted in triplicate. Different letters (a–e) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Co-Culture Assay, Positive Control, Concentration Assay

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Evaluation of iNOS protein levels in differentiated Caco-2 cells and NOx release into the apical medium. Caco-2 cells were pre-treated for 90 min with BFM (1/20, v / v ) and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. ( A ) Representative immunoblot (IB) images and ( B ) densitometric analysis of iNOS levels normalized to β-Actin. ( C ) NOx concentration. Data are expressed as mean ± SD ( n = 3). Different letters (a–d) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Positive Control, Concentration Assay, Western Blot

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Measurement of intracellular ROS production in differentiated Caco-2 cells. Caco-2 cells were pre-treated for 90 min with BFM (1/20, v / v ) and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. ( A ) Representative fluorescence histograms and ( B ) quantitative analysis of ROS generation. Data are expressed as mean ± SD of three experiments conducted in triplicate. Different letters (a–e) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Positive Control, Concentration Assay, Fluorescence

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Assessment of occludin protein levels in differentiated Caco-2 cells. Caco-2 cells were pre-treated for 90 min with BFM (1/20, v / v ) and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. ( A ) Representative immunoblot (IB) images and ( B ) densitometric analysis of occludin levels normalized to β-Actin. Data are expressed as mean ± SD ( n = 3). Different letters (a–d) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Positive Control, Concentration Assay, Western Blot

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Evaluation of NF-κB p65 nuclear translocation levels in RAW264.7 cells. Caco-2 cells were pre-treated for 90 min with BFM (1/20, v / v ) and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. ( A ) Representative immunoblot (IB) images and ( B , C ) densitometric analysis of NF-κB p65 levels in cytosolic and nuclear fractions, normalized to β-Actin and Lamin B1, respectively. Data are expressed as mean ± SD ( n = 3). Different letters (a–d) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Translocation Assay, Positive Control, Concentration Assay, Western Blot

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Assessment of COX-2 protein levels in RAW264.7 cells and PGE 2 secretion into the basolateral medium. Caco-2 cells were pre-treated for 90 min with BFM (1/20, v / v ) and budesonide (1 µM, positive control), as indicated by (+), while absence of treatment is indicated by (−). Subsequently, RAW264.7 macrophages were stimulated with LPS at a concentration of 1 µg/mL for 24 h. ( A ) Representatives immunoblot (IB) images and ( B ) densitometric analysis of COX-2 levels normalized to β-Actin. ( C ) PGE 2 concentration. Data are expressed as mean ± SD ( n = 3). Different letters (a–d) indicate statistically significant differences among all groups within each panel ( p < 0.05) (one-way ANOVA followed by Tukey’s test).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Positive Control, Concentration Assay, Western Blot

Journal: Antioxidants

Article Title: Anti-Inflammatory Activity of In Vitro Digested Manna in a Caco-2 and RAW264.7 Cells Co-Culture Model of Inflammatory Bowel Disease

doi: 10.3390/antiox15050601

Figure Lengend Snippet: Evaluation of pharmacological interactions between budesonide and BFM using CI analysis on different inflammatory markers. Measurements were performed in the apical medium (differentiated Caco-2 cells: IL-8, NOx; RAW264.7 macrophages: IL-6, TNF-α, PGE 2 ) and intracellularly (differentiated Caco-2 cells: iNOS, ROS, occludin; RAW264.7 macrophages: NF-κB p65, COX-2). A CI ratio less than 1 indicates antagonism (–), equal to 1 represents additivity (+), and greater than 1 demonstrates synergy (++). Data are expressed as mean ± SD ( n = 3).

Article Snippet: Human colonic epithelial adenocarcinoma (Caco-2) cells and murine macrophage/monocyte (RAW264.7) cell lines were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: