Streptozotocin (stz)

Six-week-old C57BL/6 background ApoE^−/− deficit mice purchased from Vital River (Beijing, China) were employed to establish the T2DM AS model according to previous descriptions with several modifications [13 (link), 14 (link)]. Animals were maintained in independent polypropylene cages in controlled ambient conditions and were free to water and standard chow. Mice were weaned for 2 weeks. Intraperitoneal injections of streptozotocin (STZ, 55 mg/kg bodyweight, Sigma-Aldrich) were administrated to mice for 5 consecutive days. Then, the animals were fed Western diet (21% milk fat and 0.15% total cholesterol, TD.88137, Harlan Teklad) for 16 weeks. Control rats received intraperitoneal injections of vehicle buffer and fed Western diet. An automatic blood glucose analyzer (Roche) was used to determine fasting blood glucose (FBP) in blood sampled from the tail vein 6 weeks after STZ injections. The TLR4 inhibitor was administrated to animals according to previous descriptions with modification [15 (link)]. DM AS animals were pretreated with TAK-242 (MedChemExpress, Shanghai, China) and twice a week for 16 weeks after modeling by intraperitoneal injection (dissolved in DMSO, 3 mg/kg bodyweight) prior to STZ injections. Animal experimental protocols were reviewed and approved by Medical Research Ethics Committee of Shaanxi Provincial People's Hospital.

Animal experiments were approved by the Institutional Animal Care and Use Committee of Kyungpook National University. C57BL/6J (male, 8- to 9-week-old) mice were obtained from Central Lab Animal Inc. Mice were injected intraperitoneally with STZ (50 mg/kg; Sigma-Aldrich, dissolved in 0.1 M citrate buffer, pH 4.5). Fasting blood glucose levels were monitored on day 6 after STZ injection, and mice with fasting blood glucose levels of >300 mg/dl were included in this study. Excisional dorsal full-thickness skin wounds (6 mm in diameter) were induced in the center of each dorsal skin on day 7 after STZ injection, and mice were administered either vehicle or bempedoic acid (10 mg/kg; MedChemExpress, dissolved in corn oil) on every 2 d. The wound area ratio was quantified by calculating the percentage of wound area compared to initial wound area.

Streptozotocin (STZ, purity > 98.0%, No. 18883–66-4), Celastrol (purity > 99.9%, No. 34157–83-0), and LY294002 (purity > 99.9%, No. 154447–36-6) were purchased from MedChem Express (Shanghai, China). The anesthetic was sodium pentobarbital (Sigma-Aldrich, St. Louis, MO, USA). LC3B (No. 2775 s) and β-actin (No. YM3028) antibodies were obtained from Auragene Cell Signaling Technology Company (Hunan, China). p-AKT (No. AM1009), AKT antibody (No. AM2058) were purchased from ABZOOM (China). Antibody against PI3K (No. ab151549) and GAPDH (No. YM3029) were respectively purchased from Abcam (UK), Immunoway (China). The commercial assay 93 kit (MeiKang, Ningbo, China) used to measure blood urea nitrogen (BUN), serum creatinine (Scr), and urine protein was purchased from the Hunan SJA Laboratory Animal Co., Ltd. (Hunan, China). TGF-β1 ELISA kits were bought from the Nanjing Jiancheng Institute of Biotechnology (Nanjing, China).