Anti-obesity effect of unsaponifiable matter from hemp seed in 3T3-L1 adipocytes and high-fat diet-induced obese mice

The favorable lipid profile of hemp seed could be a potential source of unsaponifiable matter rich in fat-soluble phytochemicals such as phytosterols, vitamin E, and cannabidiol (CBD). Despite its nutritional value, studies investigating the functional properties of hemp seed, particularly its anti-obesity potential, remain limited. The aim of this study was to obtain unsaponifiable matter from hemp seed (HUSM), analyze its fat-soluble phytochemicals and evaluate its anti-obesity activity using both <i>in vitro</i> and <i>in vivo</i> experimental models. The results showed that HUSM contained abundant carotenoids, vitamin E, phytosterols, policosanols, and CBD, with trace amounts of THC (0.06%), Furthermore, HUSM inhibited adipocyte differentiation and lipid accumulation in a dose-dependent manner, significantly reducing lipid accumulation by up to 79% without cytotoxicity in 3T3-L1 adipocytes. HUSM treatment led to reduced abdominal size and body weight gain, decreased adipose tissue and liver size, and lower plasma triglycerides, total cholesterol, and LDL cholesterol levels. These effects were mediated through the AMPK signalling pathway, which plays a pivotal role in regulating adipogenesis and lipogenesis. Additionally, HUSM improved adipokine balance, reducing leptin and increasing adiponectin levels, indicating recovery of dysfunctional adipose tissues. These findings highlight the potential of HUSM as a natural anti-obesity therapeutic, offering new avenues for the treatment and prevention of obesity and related metabolic disorders through the AMPK signalling pathway.