Metabolic characteristics of mice under HFHSD feeding.

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The characteristics of mice are summarized in Table 1. Four weeks feeding of C57Bl/6 mice with the HFHSD resulted in significant increase in body weight (20%, p<0.001) and epidydimal adipose tissue weight (336%, p<0.001), as compared to the standard diet (SD)-fed mice. Plasma glucose, FFA and triglyceride levels were similar in both groups of mice, whereas plasma insulin (72%, p<0.05) and leptin (79%, p<0.05) levels were increased in HFHSD mice compared to SD mice. After 16 weeks of diet, body weight and epidydimal fat weight gains in HFHSD animals were more marked. In addition, 16 week HFHSD mice were clearly hyperglycaemic (p<0.001) and hyperinsulinaemic (p<0.001), compared to 16 week SD mice. Plasma glucose levels of 16 week HFHSD mice were significantly higher (p<0.001) than 4 week HFHSD mice. Plasma leptin (p<0.001), FFA (p<0.05) and triglyceride levels (p<0.001) were all increased after 16 weeks of HFHSD feeding compared to SD mice.

Glucose and insulin tolerance tests showed that 4 week HFHSD mice were glucose intolerant, while their response to insulin injection remained unaltered compared to SD mice (Figures S1A and S1B). In contrast, after 16 weeks of HFHSD feeding, mice presented an altered response to both glucose and insulin injection compared to SD mice, indicating that 16 week HFHSD mice are insulin resistant (Figures S1A and S1B). Decreased insulin responsiveness in 16 week HFHSD mice was associated with intramyocellular lipid accumulation (Figure S2A), increased basal IRS-1 serine phosphorylation at Ser632 (45%, p<0.05, Figure S2B) and with a decrease of ex vivo insulin-stimulated Akt phosphorylation at Ser473 (80%, p<0.01, Figure S2C) in gastrocnemius muscle. In contrast, insulin-stimulated Ser473 phosphorylation of Akt was not modified in 4 week HFHSD mice compared to SD mice (Figure S2C).