ApoE-/- Mice Model of T2DM and Atherosclerosis

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.