Journal: International Journal of Molecular Sciences

Article Title: Novel Benzoxazoles Containing 4-Amino-Butanamide Moiety Inhibited LPS-Induced Inflammation by Modulating IL-6 or IL-1β mRNA Expression

doi: 10.3390/ijms23105331

Figure Lengend Snippet: Compounds 5f and 4d attenuate inflammation in vitro. Effects of compounds (10 µM) on the protein levels of STAT3, IκB, and NF-κB in AML-12 cells. ( a ) Expression levels of inflammation related-proteins in vitro. ( b – d ) Protein expression levels were normal-ized against the indicated protein. * p < 0.05 compared with the group treated with the PBS (vehicle). + p < 0.05 compared with the group treated with the LPS. Data are pre-sented as the mean ± SD.

Article Snippet: The human keratinocytes HaCaT or the alpha mouse liver 12 (AML-12) cells were obtained from Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: In Vitro, Expressing

Journal: Cells

Article Title: Ursodeoxycholic Acid Regulates Hepatic Energy Homeostasis and White Adipose Tissue Macrophages Polarization in Leptin-Deficiency Obese Mice

doi: 10.3390/cells8030253

Figure Lengend Snippet: Ursodeoxycholic acid (UDCA) alleviates high free fatty acid (HFFA)-induced hepatocyte lipogenesis, reactive oxygen species (ROS) production, and mitochondrial dysfunction in AML12 cells. AML12 cells were treated with 1 mM HFFA with 10, 30, 100 μM UDCA. ( A ) Lipid accumulation display using Oil Red O stain (red). ROS levels were measured using DCFH-DA (green) stain. Images of AML12 cells stained with Mito Tracker for mitochondria (red). qRT-PCR analysis of ( B ) Complex I, II, III, IV, and V mRNA expression in AML12 cells. Relative mRNA expression was normalized to Gapdh and then normalized to the controls. ( C ) Immunofluorescence analysis of SREBP1c (green), CD36 (red), NF-κB (green), and FXR (green) expression, and DAPI (blue) for nuclear. Scale bar, 25 μm. qRT-PCR analysis of ( D ) Srebp-1c, Fas , and Scd-1 mRNA expression in AML12 cells. In all panels, results are expressed as the mean ± S.E.M. of five independent experiments, and statistical significance of differences between means was assessed using an unpaired Student’s t -test (* p ≤ 0.05; 0 mM HFFA vs. 1 mM HFFA. # p ≤ 0.05; 1 mM HFFA vs. 1 mM HFFA+ 100 μM UDCA). UDCA, ursodeoxycholic acid; HFFA, high free fatty acid; ROS, reactive oxygen species; SREBP-1c, sterol regulatory element-binding protein-1c; CD36, cluster of differentiation 36; NF-κB, nuclear factor kappa-light-chain-enhancer of activated B cells; FXR, farnesoid X receptor; Fas , fatty acid synthase; Scd-1, stearoyl-CoA desaturase-1; qRT-PCR, quantitative real-time polymerase chain reaction; Gapdh , glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Alpha mouse liver 12 (AML12) cell line (passages 7–10, obtained from Bioresource Collection and Research Centre, Taiwan) was grown at 37 °C in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 (DMEM/F12), 10% fetal bovine serum (FBS), 2 mmol/L l -Glutamine and 100 μg/mL penicillin and streptomycin (both from Gibco, AntiSel, Greece) at 5% CO 2 .

Techniques: Staining, Quantitative RT-PCR, Expressing, Immunofluorescence, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Heliyon

Article Title: Oral administration of kynurenic acid delays the onset of type 2 diabetes in Goto-Kakizaki rats

doi: 10.1016/j.heliyon.2023.e17733

Figure Lengend Snippet: KYNA enhances mRNA and protein expression of hepatic UCP genes. (A, B) Real-time quantitative PCR analysis (n = 6) of gene expression in HepG2 cells after 48 h of 10 μmol/L (K10) or 100 μmol/L (K100) KYNA treatment compared with control group (white punctuated line). (C, D) Western blot analysis of UCP2 expression in HepG2 cells after 48 h of 100 μmol/L KYNA treatment (n = 3), (E, F) Real-time quantitative PCR analysis of gene expression in HepG2 cells after 2 h treatment with 10 μmol/L (K10) or 100 μmol/L (K100) KYNA (n = 6), (G) Quantitative PCR analysis of gene expression in AML-12 cells after 48 h treatment with 10 μmol/L (K10) or 100 μmol/L (K100) KYNA (n = 6), (H) Expression of hepatic energy metabolism genes in GK rats after oral administration 25 mg/kg KYNA (KYNA) or NaCl (Ctrl) (I) Comprehensive analysis of transcriptomic pathways in KYNA-treated HepG2 cells after 48 h KYNA, 100 μmol/L (n = 3). Statistical analysis using t -test, * p < 0.05.

Article Snippet: HepG2 human liver cells and AML-12 mouse liver cells (Procell, China) were cultured in MEM basal medium (Gibco, China) containing 10% fetal bovine plasma (Gibco, Australia).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot