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For mitochondrial ROS detection, the cells were stained with 5 g/mL MitoSox dye (Thermo Scientific, Waltham, MA, USA) at 37 C for 15 min, followed by three PBS washes

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2) Under conditions of sustained systemic inflammation (as seen in PEM), high levels of circulating cytokines may degrade tight junction proteins (e.g., claudin-5 and occludin), leading to BBB disruption and increased paracellular permeability

Height, weight, and body mass index (BMI) were measured at baseline and the end of the study