Abstract

Hepatic organoids successfully modeled native liver complexity by maintaining core hepatocyte functions and upregulating key metabolic and repair genes. • C3G exerted the weakest lipid-lowering effect, primarily by alleviating oxidative stress. • P3G showed moderate efficacy, mainly through suppressing lipogenesis. • D3G showed the most potent lipid-lowering activity, largely attributed to its anti-inflammatory effects and regulation of the PPAR pathway. Organoids represent promising models for studying lipid deposition due to their physiological relevance to native organs. Although studies indicate that anthocyanin (ACNs) mixtures containing Cyanidin-3-O-glucoside (C3G), Pelargonidin-3-glucoside (P3G), and Delphinidin-3-O-glucoside (D3G) alleviate hepatic lipid deposition, comparative analyses of their mechanistic underpinnings remain limited. In this study, 3D hepatic organoids were cultured, exhibiting key functional characteristics and up-regulated expression of lipid metabolism-associated genes. Oil Red O staining revealed that P3G and D3G exhibit comparable efficacy in reducing lipid deposition, and both were more effective than C3G. Mechanistically, D3G significantly reduced IL-6 and TNF-α levels while elevating IL-10, whereas C3G substantially elevated glutathione levels. Gene and protein expression profiling demonstrated that P3G exhibited the most significant suppression of SREBP-1 expression, whereas D3G induced activation of the AMPK signaling pathway. Moreover, D3G promoted lipid oxidation by specifically up-regulating CPT-1 A and ACOX1 protein expression. Furthermore, compared to C3G and P3G, D3G significantly up-regulated the expressions of PPARα and PPARγ. These findings suggest that D3G, among the three anthocyanins, exhibited the most potent hypolipidemic effect, which was attributable to its anti-inflammatory, antioxidant, and PPAR pathway-regulating activities. Comparative analysis of three anthocyanins for mitigating lipid accumulation in hepa