# Anti-Obesity Effects of Acid-Processed Citrus reticulata Blanco Peel Extract Enriched in Highly Bioactive Polymethoxyflavones: Inhibition of 3T3-L1 Adipocyte Differentiation and Therapeutic Efficacy in ob/ob Mice

## TL;DR

This study shows that an acid-treated citrus peel extract can reduce fat cell formation and obesity in mice, suggesting it could be a natural treatment for obesity.

## Contribution

The novel contribution is the acid-processing method that enhances the bioactivity of polymethoxyflavones in citrus peel extract for anti-obesity effects.

## Key findings

- Acid treatment increased levels of bioactive polymethoxyflavones like 5-demethylnobiletin and 5-demethyltangeretin in citrus peel extract.
- CRBE inhibited 3T3-L1 adipocyte differentiation and reduced body weight gain in ob/ob mice without affecting food intake.
- The extract improved liver function and lipid profiles in obese mice.

## Abstract

Background/Objectives: Chronic diseases linked to obesity represent a major global health challenge. Although pharmaceutical treatments show efficacy, their use is often limited by side effects. Methods: This study investigated the anti-obesity effects of acid-processed Citrus reticulata Blanco peels extract (CRBE) prepared through reflux extraction with 50% ethanol, followed by acid treatment using 3 M hydrogen chloride and neutralization. Results: Following acid treatment, the composition of the extract showed a marked increase in the 5-demethylated forms of polymethoxyflavones, particularly 5-demethylnobiletin (31.86 mg/g) and 5-demethyltangeretin (34.68 mg/g), whereas the concentrations of the typical citrus polymethoxyflavones, nobiletin (14.82 mg/g) and tangeretin (10.61 mg/g), decreased. Using 3T3-L1 preadipocytes, CRBE inhibited adipogenesis concentration dependently, substantially decreasing the expression of adipogenic transcription factors and lipid metabolism-related proteins. In ob/ob mice, oral CRBE substantially suppressed body weight gain without affecting food intake, while normalizing liver function indicators and improving serum lipid profiles by reducing total cholesterol, triglycerides, and low-density lipoprotein. Conclusions: Acid-processed CRBE effectively inhibits adipocyte differentiation and exhibits anti-obesity effects in vivo, offering potential as a natural agent for obesity management with minimal side effects.