Study on the Effect of Koumiss Extract in Alleviating Non‐Alcoholic Fatty Liver Disease in Zebrafish Model by Improving Mitochondrial Function and Inhibiting Fat Deposition

Non-alcoholic fatty liver disease (NAFLD) has emerged as a significant health issue due to the pathological accumulation of fat in the liver in the absence of excessive alcohol intake, with mitochondrial dysfunction being a critical underlying mechanism. This study aimed to evaluate the therapeutic potential of koumiss extract, along with 2-furanic acid and α, α-trehalose, in modulating mitochondrial function and mitigating fat deposition in NAFLD. Utilizing molecular docking techniques, we assessed the binding affinities of these compounds to mitochondrial complex I assembly (MCIA) proteins, while establishing both in vitro (HepG2 cell line) and in vivo (zebrafish model) NAFLD models to measure lipid accumulation and related biochemical parameters, including triglyceride (TG), total cholesterol (TC), and lactate dehydrogenase (LDH) levels, alongside the expression profiles of MCIA proteins. Our results demonstrated that koumiss extract, 2-furanic acid, and α, α-trehalose significantly decreased TG and LDH levels indicative of steatosis in HepG2 cells, while also reducing the expression of MCIA-related proteins. In vivo experiments using a zebrafish NAFLD model demonstrated pronounced liver steatosis in the model group. Treatment with koumiss extract, 2-furanic acid, and α, α-trehalose significantly alleviated liver steatosis and reduced TG and TC levels. Furthermore, mRNA expression levels of ACAD9, ECSIT, NDUFAF1, and NDUFAF2 were significantly downregulated in the treatment groups. Koumiss extract, 2-furanic acid, and α, α-trehalose exhibit significant effects in reducing MCIA-related proteins and steatosis in NAFLD models. Consequently, these results suggest that koumiss extract and its analogs hold promise as therapeutic agents for NAFLD, potentially enhancing liver lipid homeostasis.