Integration of network pharmacology, deep learning, and molecular biology reveals the efficacy of Citrus aurantium L. var. amara Engl. blossom extract in ameliorating diabetic osteoporosis

  • J Ethnopharmacol. 2026 Oct 28:369:121939. doi: 10.1016/j.jep.2026.121939.
Qian Ni  1 ,  Yan-Ling Zhang  1 ,  Wen-Xin Li  1 ,  Wei-Feng Cai  1 ,  Qi-Cong Chen  1 ,  Wei-Le Liu  2 ,  Cui-Ping Jiang  3 ,  Shao-Wei Zheng  4 ,  Chun-Yan Shen  5
Affiliations
  • 1. School of Traditional Chinese Medicine, Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Southern Medical University, Guangzhou, 510515, China.
  • 2. Department of Orthopaedics, Huizhou First Hospital, Guangdong Medical University, Huizhou, Guangdong, 516003, China.
  • 3. School of Traditional Chinese Medicine, Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Southern Medical University, Guangzhou, 510515, China. Electronic address: [email protected].
  • 4. Department of Sports Medicine and Rehabilitation, Shenzhen Xinhua Hospital, Peking University Shenzhen Hospital, Shenzhen, 518131, China; Department of Orthopaedics, Huizhou First Hospital, Guangdong Medical University, Huizhou, Guangdong, 516003, China. Electronic address: [email protected].
  • 5. School of Traditional Chinese Medicine, Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Southern Medical University, Guangzhou, 510515, China. Electronic address: [email protected].
PMID: 42217588 DOI: 10.1016/j.jep.2026.121939
Abstract

Ethnopharmacological relevance: Diabetic osteoporosis (DOP) is a type of metabolic bone disease. Blossom of Citrus aurantium L. var. amara Engl. (CAVA), a medicinal and edible herb widely employed in China which can alleviate indigestion and replenish qi, has been demonstrated to be effective in treating metabolic disorders such as obesity and diabetes. However, its effects on DOP remains uncertain.

Aim of the study: To explore the protective effects and mechanisms of blossom of CAVA against DOP.

Materials and methods: Ethanol extract of blossom of CAVA (CAVAE) was characterized using HPLC/MS. Network pharmacology, deep learning, molecular docking and molecular dynamics simulations were employed to predict key constituents and possible pathways. High-fat diet and streptozocin-induced type 2 diabetes mellitus (T2DM) mice were established and Micro-CT, X-ray imaging, morphological analysis and Molecular Biology assays were performed to investigate the effects and underlying mechanisms of CAVAE on DOP.

Results: The 44 compounds identified by HPLC/MS analysis suggest that CAVAE may prevent DOP via PI3K/Akt and Wnt/β-catenin pathways. Experimental validation demonstrated that CAVAE significantly improved diabetes and osteoporosis in T2DM mice, as evidenced by the decreased blood glucose content, increased bone mineral density, enhanced trabecular bone number, improved femoral bone microstructure, restored femur stability, bone metabolism and reduced serum calcium ion levels, etc. Mechanistic investigations further confirmed that CAVAE probably alleviated DOP in T2DM mice by modulating PI3K/Akt and Wnt-3a/β-catenin pathways.

Conclusion: CAVAE might exert anti-osteoporotic effects on T2DM mice through concurrent activation of PI3K/Akt and Wnt-3a/β-catenin pathways.

Keywords
Citrus aurantium L. var. amara Engl.; Diabetic osteoporosis; Type 2 diabetes mellitus.