Journal: iMeta

Article Title: A gut microbiome‐lipid axis in early pregnancy is associated with metabolic dysregulation and diabetes risk

doi: 10.1002/imt2.70166

Figure Lengend Snippet: Experimental evaluation of a prioritized Ruminococcus bicirculans ‐glycosphingolipid axis. (A) Production of LacCer 24:1 after anaerobic incubation of R. bicirculans under different substrate conditions. (B) Dose‐dependent production of LacCer 24:1 by R. bicirculans in the presence of increasing concentrations of glucosylceramide (GlcCer) 24:1, with or without d ‐galactose supplementation. (C) Schematic overview of the in vivo intervention experiment. Mice were subjected to high‐fat diet feeding, streptozotocin (STZ) treatment, antibiotic‐mediated microbiota depletion, and subsequent intervention with vehicle, R. bicirculans or LacCer 24:1. (D) Insulin tolerance test (ITT) curves and corresponding area under the curve (AUC) in mice following LacCer 24:1 administration. (E) ITT curves and corresponding AUC in mice following R. bicirculans colonization. AUCs were calculated using the trapezoidal rule. (F) Representative Western blots showing the time course of insulin‐stimulated AKT phosphorylation in primary hepatocytes pretreated with LacCer 24:1 or vehicle. (G) Schematic model of the proposed R. bicirculans ‐glycosphingolipid‐host interaction axis. R. bicirculans may contribute to LacCer‐related metabolite production, which is associated with modulation of insulin‐stimulated AKT signaling in hepatocytes. Data are presented as mean ± SEM where applicable. Statistical significance was assessed using two‐sided tests as indicated * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. The schematic was created with BioRender.com. ABX, antibiotics; d ‐Gal, d ‐galactose; DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; GlcCer, glucosylceramide; HFD, high‐fat diet; ITT, insulin tolerance test; pAKT, phosphorylated AKT; PBS, phosphate‐buffered saline; SEM, standard error of the mean; STZ, streptozotocin.

Article Snippet: Mice were fed a high‐fat diet (HFD; XTHF60‐1, Jiangsu Xietong Pharmaceutical Bioengineering Co., Ltd.; 60% kcal from fat) for 8 weeks, followed by intraperitoneal injection of STZ (MedChemExpress, Cat# HY‐13753; 40 mg/kg) for 5 consecutive days to induce metabolic dysfunction.

Techniques: Incubation, In Vivo, Western Blot, Phospho-proteomics, Saline

Journal: Diabetes & Metabolism Journal

Article Title: Abnormally Elevated PKCδ Delays Diabetic Wound Healing by Inhibiting the GAD1-GABA Pathway

doi: 10.4093/dmj.2024.0450

Figure Lengend Snippet: Decreased gamma-aminobutyric acid (GABA) levels and glutamate decarboxylase 1 (GAD1) expression in high glucose (HG)-treated human umbilical vein endothelial cells and skin tissues of diabetic mice. (A) Measurement of GABA concentration in culture medium 72 hours after HG treatment. (B) Quantitative polymerase chain reaction assay of GAD1 mRNA level 72 hours after HG treatment. (C, D) Western blot assay and quantification of GAD1 protein level 72 hours after HG treatment. (E, F) Immunofluorescence assay and quantification of GAD1 protein level 72 hours after HG treatment. The scale bar is 20 μm. (G) Measurement of GABA concentration in the serum of streptozotocin (STZ)-induced type 2 diabetes mellitus (T2DM) mice. (H) Measurement of GABA concentration in the skin of STZ-induced T2DM mice. (I, J) mRNA and protein levels of GAD1 in the skin of STZ-induced T2DM mice. Each experiment was replicated for thrice and representative images are shown. Quantification was performed using a two-tailed unpaired Student’s t -test. The results are presented as the mean±standard error of the mean. NG, normal glucose; DAPI, 4′,6-diamidino-2-phenylindole; Veh, vehicle. a P <0.01, b P <0.001 vs. controls.

Article Snippet: The chemicals used in this study included: γ-aminobutyric acid (#HY-N0067, MedChemExpress, Monmouth Junction, NJ, USA), 3-mercaptopropionic acid (3-MPA; #M5801, Sigma, St. Louis, MO, USA), and streptozocin (#HY-13753, MedChemExpress).

Techniques: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Two Tailed Test