Journal: Pharmaceuticals

Article Title: The Potential of Epigallocatechin-3-gallate (EGCG) as Complementary Medicine for the Treatment of Inflammatory Bowel Disease

doi: 10.3390/ph16050748

Figure Lengend Snippet: Effect of LPS stimulation, 2 µg/mL, and 200 µg/mL EGCG treatment on the cytokines released by the MDM in the co-culture setup in comparison to the healthy state (medium control). The stimulation with LPS led to an increase in the tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-6, and IL-8 release. No significant effect of the treatment with 2 µg/mL EGCG was observed. After the treatment with 200 µg/mL EGCG, no IL-6 release was detectable, and a significant decrease in the IL-8 release was achieved. Results are displayed as mean ± standard deviation for in summary n = 9 wells for each group (except for LPS (TNF-α) n = 6 wells). Samples out of three biological replicates were utilized. * p < 0.05, ** p < 0.01, and *** p < 0.001 imply a significant difference.

Article Snippet: For performing ELISA to measure the cytokine release by the MDM in the co-culture system, the Human Uncoated ELISA Kits for the tested cytokines (TNF-α, IL-6, IL-8, and IL-10) from Thermo Fisher Scientific were used and carried out according to the provided protocol.

Techniques: Co-Culture Assay, Standard Deviation

Journal: BMC Medicine

Article Title: Gut taste receptor type 1 member 3 is an intrinsic regulator of Western diet-induced intestinal inflammation

doi: 10.1186/s12916-023-02848-0

Figure Lengend Snippet: A combination of glucose, fructose, and palmitate ligands induces production and secretion of pro-inflammatory cytokines and chemokine in EECs via TAS1R3 activation. a qRT-PCR of relative mRNA expression of intestinal TAS1R3 in NCI-H716 cells after 12 h of treatment with individual or combined glucose, fructose, and palmitate to activate TAS1R3 ( n = 3/group). Time course of ( b ) GLP1 mRNA expression, c active GLP-1 levels, d IL1B mRNA expression, e IL6 mRNA expression, f TNFɑ mRNA expression, and g MCP1 mRNA expression in NCI-H716 cells after stimulation with individual or combined glucose, fructose, and palmitate ( n = 3/group). h–l NCI-H716 cells were transfected with TAS1R3 (10 nM) or scrambled control (10 nM) siRNA for 48 h and then stimulated with or without fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h ( n = 3/group). Evaluation of TAS1R3 siRNA h mRNA and i protein knockdown in EECs. Transfection with TAS1R3 siRNA inhibited TAS1R3 mRNA and protein expression in differentiated NCI-H716 cells. The collected supernatants were assayed for active j GLP-1, k IL-8, and l TNF-α levels. m–o NCI-H716 cells were pretreated with the TAS1R3 antagonist, lactisole (2.5 mM), for 30 min and then stimulated with fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h ( n = 3/group). The collected supernatants were assayed by active m GLP-1, n IL-8, and o TNF-α enzyme-linked immunosorbent assay. Data represent means ± standard errors of the mean of three to five independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 (one-way analysis of variance followed by Bonferroni post-hoc test). EEC, enteroendocrine cell; GLP-1, glucagon-like peptide 1; IL, interleukin; MCP-1, monocyte chemoattractant protein-1; qRT-PCR, quantitative reverse transcription PCR; TAS1R3, taste receptor type 1 member 3; TNFα, tumor necrosis factor-alpha; WD, western diet

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Kits specific for human TNF-α (Cat# ab181421) and IL-8 (Cat# ab46032) were purchased from Abcam.

Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: BMC Medicine

Article Title: Gut taste receptor type 1 member 3 is an intrinsic regulator of Western diet-induced intestinal inflammation

doi: 10.1186/s12916-023-02848-0

Figure Lengend Snippet: TAS1R3 controls pro-inflammatory cytokine production and secretion by regulating the mTOR–PPARγ axis. Western blotting of ( a ) phospho-mTOR, mTOR, and PPARγ expression in ND- and WD-fed Tas1r3 −/− and Tas1r3 +/+ mice and ( b ) and ( c ) densitometric data ( n = 6 mice/group). Representative images of at least three different blots and α-tubulin. d–f NCI-H716 cells were transfected with TAS1R3 (10 nM) or scrambled control (10 nM) siRNA for 48 h and stimulated with or without fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h. qRT-PCR analysis of ( d ) MTOR and ( e ) PPARG mRNA expression relative to GAPDH ( n = 5/group). g–i NCI-H716 cells were pretreated with the TAS1R3 antagonist, lactisole (2.5 mM), for 30 min and stimulated with fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h. qRT-PCR analysis of ( g ) MTOR and ( h ) PPARG mRNA expression relative to GAPDH ( n = 5/group). f and i PPARγ protein abundance was assessed by western blotting ( n = 6/group). Alpha-tubulin was used as the loading control. j–m NCI-H716 cells were transfected with TAS1R3 (10 nM) or scrambled control (10 nM) siRNA for 48 h and stimulated with the PPARγ antagonist, GW9662 (10 μM), in the presence of fructose (10 mM), glucose (10 mM) and palmitate (10 μM) for 12 h. qRT-PCR analysis of ( j ) TNFA and ( k ) IL8 mRNA expression relative to GAPDH ( n = 5/group) and enzyme-linked immunosorbent assay of ( l ) TNF-α and ( m ) IL-8 protein abundance ( n = 3/group). Data represent means ± standard errors of the mean of three to five independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 (one-way analysis of variance followed by Bonferroni post-hoc test). ND, normal diet; qRT-PCR, quantitative reverse transcription PCR; TAS1R3, taste receptor type 1 member 3; WD, Western diet

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Kits specific for human TNF-α (Cat# ab181421) and IL-8 (Cat# ab46032) were purchased from Abcam.

Techniques: Western Blot, Expressing, Transfection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay