Boiogito Ameliorates Hydrogen Peroxide-Induced Impairment of Glucose Uptake in 3T3-L1 Adipocytes

Chronic inflammation and oxidative stress in obese adipose tissue are pivotal drivers of insulin resistance and metabolic dysfunction. We previously demonstrated that the Kampo formula Boiogito (BOT) ameliorates impaired adipocyte differentiation and inflammation by inhibiting the nuclear factor kappa B (NF-κB) pathway in a tumor necrosis factor–alpha (TNF-α)-induced model. However, the specific effects of BOT on oxidative stress and glucose uptake remain to be elucidated. This study aimed to elucidate whether BOT could ameliorate hydrogen peroxide (H2O2)-induced oxidative stress, inflammation, and cellular dysfunction in 3T3-L1 adipocytes, thereby restoring impaired glucose uptake capacity. Differentiating and mature 3T3-L1 adipocytes were treated with 200 µM H2O2 alone or in combination with BOT (0.25 and 0.5 mg/mL). Oxidative stress markers (reactive oxygen species [ROS] and superoxide dismutase [SOD]), inflammatory cytokines (interleukin-6 [IL-6], monocyte chemoattractant protein-1 [MCP-1], and TNF-α), differentiation/functional markers (peroxisome proliferator-activated receptor gamma [Pparγ] and CCAAT/enhancer-binding protein alpha [C/ebpα] mRNA expression, adiponectin secretion, and intracellular lipid content), and glucose uptake (via a 2-deoxy-D-glucose [2-DG] assay) were evaluated. In both stages of adipocytes, H2O2 exposure significantly increased ROS levels and decreased SOD activity, and elevated extracellular IL-6, MCP-1, and TNF-α concentrations. These alterations were significantly attenuated by BOT co-treatment. Furthermore, BOT prevented the H2O2-induced downregulation of Pparγ and C/ebpα mRNA expression, thereby restoring adiponectin secretion and intracellular lipid content. Crucially, the marked impairment of 2-DG uptake caused by H2O2-mediated metabolic stress was significantly reversed by BOT co-treatment. These findings suggest that BOT exerts multi-targeted protective effects in 3T3-L1 adipocytes by mitigating oxidative stress, suppressing inflammation, and preserving adipocyte differentiation and function, thereby restoring impaired glucose uptake. Our study provides new insights into the therapeutic potential of BOT against insulin resistance and associated metabolic disorders.