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Rats were placed individually in sterile metabolic cages, and feces (≥500 mg per rat) were gathered into sterile, nuclease-free tubes, rapidly frozen in liquid nitrogen, and stored at −80 °C for 16S rRNA gene sequencing to analyze gut microbiota

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Furthermore, the compensatory hyperinsulinemia resulting from adipose dysfunction stimulates de novo lipogenesis in hepatocytes while simultaneously impairing muscle glucose uptake, creating a pathological metabolic state that propagates systemic insulin resistance (157)