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Immunofluorescence intensity for GFAP, AQP4, and Cx43 was quantified in manually selected regions of interest (ventromedial hypothalamic nucleus) using mean brightness values in 256 grayscale levels of 8-bit images

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MASLD is diagnosed when hepatic steatosis occurs in combination with cardiometabolic risk factors, including overweight/obesity, elevated blood pressure, atherogenic dyslipidemia, prediabetes, or T2DM (Targher et al., 2021[75], 2025[79]