Two natural products from the seeds of Citrus reticulata Blanco (Rutaceae) inhibit estrogen biosynthesis by regulating the PI3K-aromatase pathway

Xing Yang; Xi Chen; Chunqiao Shi; Qian Zhang; Qian Liu; Chunyan Zhou; Fawu Dong; Jinsong Su; Deming Liu; Yi Zhang

doi:10.3389/fphar.2025.1583409

Two natural products from the seeds of Citrus reticulata Blanco (Rutaceae) inhibit estrogen biosynthesis by regulating the PI3K-aromatase pathway

Front Pharmacol. 2025 Jun 12:16:1583409. doi: 10.3389/fphar.2025.1583409. eCollection 2025.

Authors

Xing Yang^#¹, Xi Chen^#², Chunqiao Shi¹, Qian Zhang¹, Qian Liu¹, Chunyan Zhou³, Fawu Dong⁴, Jinsong Su¹, Deming Liu², Yi Zhang¹

Affiliations

¹ Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, China.
² Chongqing Clinical Research Center for Dermatology, Chongqing Key Laboratory of Integrative Dermatology Research, Key Laboratory of External Therapies of Traditional Chinese Medicine in Eczema, Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital/the First Affiliated Hospital of Chongqing College of Traditional Chinese Medicine, Chongqing, China.
³ Department of General Surgery, Chongqing Traditional Chinese Medicine Hospital/the First Affiliated Hospital of Chongqing College of Traditional Chinese Medicine, Chongqing, China.
⁴ Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan University of Traditional Chinese Medicine, Kunming, China.

^# Contributed equally.

Abstract

Introduction: The seeds of Citrus reticulata Blanco (Rutaceae) (SCR), a traditional Chinese medicine derived from Citrus, is known for its diverse bioactivities, including potential anti-breast cancer effects, but the mechanism of action remains unclear.

Methods: This study aims to elucidate the active ingredients of SCR and their mechanisms of action on estrogen biosynthesis. A comprehensive phytochemical analysis, employing various chromatographic techniques, led to the isolation of 26 compounds from SCR.

Results: The effects of these compounds on estrogen biosynthesis were evaluated in human ovarian granulosa-like KGN cells, which play a crucial role in the progression of hormone-dependent breast cancers. Network pharmacology analysis revealed that SCR may influence breast cancer development by modulating phosphorylation-related biological processes and the PI3K/AKT pathway. Among the isolated compounds, Callyspongidipeptide A (Calp) and Hesperetin 7-O-β-D-glucopyranoside (Hesp) exhibited significant inhibitory effects on estrogen biosynthesis. Calp and Hesp selectively regulated the expression of aromatase (Aro) PI.3 and P2 promoters via the PI3K/AKT pathway, inhibiting Aro mRNA and protein expression.

Discussion: These findings provide novel insights into the chemopreventive potential of SCR and support its role in the development of therapies aimed at reducing the risk of hormone-related cancers.

Keywords: Hesperetin 7-O-β-D-glucopyranoside; aromatase; aromatase inhibitors; callyspongidipeptide A; the seeds of Citrus reticulata Blanco (Rutaceae).