Identification of CA2 as a Potential Candidate Gene Involved in the Therapeutic Effect of Huatan Huoxue Decoction on Polycystic Ovary Syndrome: An Integrated Study of Network Pharmacology, Mendelian Randomization, and Experimental Validation

Chan Zhu, Department of Traditional Chinese Medicine, West China Second University Hospital, Sichuan University, Chengdu, China
Yijin Zhang, Department of Gynaecology, Shanghai municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Panwei Hu, Department of Gynecology and Obstetrics, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China
Cong Qi, Department of Gynecology and Obstetrics, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China
Qiu Chen, Hospital of Chengdu University of Traditional Chinese Medicine, Department of Endocrinology, Chengdu, Sichuan, China.
Hong Yang, Department of Gynaecology, Shanghai municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaFollow
Jing Li, Integrated Chinese and Western Medicine Department of Gynecology, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China

Keywords

CA2, Huatan Huoxue Decoction, Network Pharmacology, Polycystic Ovary Syndrome, Rat Model

Abstract

Purpose: Huatan Huoxue decoction (HTHXD) is effective in treating polycystic ovary syndrome (PCOS); however, its underlying mechanisms of action remain unclear. Methods: Gene targets were obtained through Mendelian randomization (MR) analysis using druggable genes from public databases and genome-wide association study data for PCOS from the FinnGen R12 database. These targets were integrated with the active ingredients and protein targets of HTHXD retrieved from the TCMSP and BATMAN-TCM databases for network pharmacology analysis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted. Core targets were validated using molecular docking. Experimental validation was performed using a letrozole-induced PCOS rat model treated with HTHXD, with key targets assessed via quantitative PCR (qPCR) and immunohistochemical staining. Results: MR analysis identified 131 proteins causally related to PCOS, of which 52 with strong associations remained after quality control. Network pharmacology analysis revealed 549 gene targets and 243 active ingredients in HTHXD. The intersection of these datasets identified four core targets for HTHXD in PCOS treatment (CA2, CASP3, SNCA, and TSPO). Molecular docking identified tanshindiol B, 4-methylenemiltirone, and danshenol B in Danshen and crocetin in leech as key active components. HTHXD substantially ameliorated polycystic ovarian morphology and reduced corpora lutea in PCOS model rats. CA2 expression was significantly upregulated in PCOS ovarian tissue (P < 0.0001), and markedly reduced after HTHXD treatment, consistent with immunohistochemical findings. Conclusion: CA2 may serve as a potential candidate gene associated with the therapeutic response to HTHXD in PCOS, with tanshindiol B and crocetin as major active components.

Abstract Image

Recommended Citation

Zhu, Chan; Zhang, Yijin; Hu, Panwei; Qi, Cong; Chen, Qiu; Yang, Hong; and Li, Jing (2026) "Identification of CA2 as a Potential Candidate Gene Involved in the Therapeutic Effect of Huatan Huoxue Decoction on Polycystic Ovary Syndrome: An Integrated Study of Network Pharmacology, Mendelian Randomization, and Experimental Validation," Journal of Food and Drug Analysis: Vol. 34 : Iss. 3 , Article 14.
Available at: https://doi.org/10.38212/2224-6614.3606