High-Fat Diet and Metabolic Syndrome Mitigation

C57BL/6 J mice were housed in a vivarium in accordance with the guidelines of the animal facility center of Kumamoto University. The light/dark cycle was set at 12 h, and the temperature was kept at a room temperature of 20 °C. To prepare diabetes model with HFD loading, an HFD purchased from Oriental Yeast Co., Ltd. (Tokyo, Japan) was used as previously described^{23 (link)}. HFD was composed of 14% lard, 14% beef tallow, 25% casein, 20% sucrose, 15% cornstarch, 5% cellulose.
Five-week-old mice were randomly divided into 4 groups and given the following diet for 10 weeks: control diet for 10 weeks (n = 6); HFD for 10 weeks (n = 6); HFD for first 6 weeks and HFD supplemented with MSE powder 500 mg/kg/day for last 4 weeks (n = 6); and HFD for the first 6 weeks and HFD supplemented with MSE powder 1,000 mg/kg/day for the last 4 weeks (n = 6). HFD was orally administered to mice. Adipose tissue, muscle and liver samples were collected from mice that completed the diet. Since MSE powder has been reported to be well tolerated at an oral administration of 5,000 mg/day in humans^{5 (link)}, the dose of 1,000 mg/kg/day for mice was calculated using a human equivalent dose (5,000 mg/60 kg/day in human)²⁴ .
All experiments on mice were approved by the Animal Welfare Committee of Kumamoto University and were conducted in adherence to institutional regulations and guidelines.