Nimbin analog N2 alleviates high testosterone induced oxidative stress in CHO cells and alters the expression of Tox3 and Dennd1a signal transduction pathway involved in the PCOS zebrafish

• Published in: Phytotherapy research 2023-04, Vol.37 (4), p.1449-1461
• Main Authors: Sudhakaran, Gokul, Rajesh, Ravi, Murugan, Raghul, Velayutham, Manikandan, Guru, Ajay, Boopathi, Seenivasan, Muthupandian, Saravanan, Gopinath, Pushparathinam, Arockiaraj, Jesu
• Format: Article
• Language: English
• Subjects: Animals; Anti-inflammatory agents; Antioxidants; Antioxidants - metabolism; Apoptosis; CHO Cells; Cricetinae; Cricetulus; Danio rerio; Death Domain Receptor Signaling Adaptor Proteins - genetics; Death Domain Receptor Signaling Adaptor Proteins - metabolism; Female; Gene expression; Genes; Guanine Nucleotide Exchange Factors - genetics; Guanine Nucleotide Exchange Factors - metabolism; Histopathology; Humans; Maturation; Molecular docking; Molecular Docking Simulation; Molecular dynamics; Neem; Nimbin; Ovaries; Oxidative Stress; polycystic ovarian syndrome; Polycystic ovary syndrome; Polycystic Ovary Syndrome - pathology; Reactive Oxygen Species - metabolism; Signal Transduction; Testosterone; Zebrafish; Zebrafish - metabolism
• ISSN: 0951-418X; 1099-1573
• DOI: 10.1002/ptr.7685

Polycystic ovarian syndrome (PCOS) is a hormonal disorder that causes enlargement of ovaries and follicular maturation arrest, which lacks efficient treatment. N2, a semi‐natural triterpenoid from the neem family, was already reported to have antioxidant and antiinflammatory properties in our previous report. This study investigated the anti‐androgenic property of N2 on testosterone‐induced oxidative stress in Chinese Hamster Ovarian cells (CHO) and PCOS zebrafish model. The testosterone exposure disrupted the antioxidant enzymes and ROS level and enhanced the apoptosis in both CHO cells and PCOS zebrafish. However, N2 significantly protected the CHO cells from ROS and apoptosis. N2 improved the Gonado somatic index (GSI) and upregulated the expression of the SOD enzyme in zebrafish ovaries. Moreover, the testosterone‐induced follicular maturation arrest was normalized by N2 treatment in histopathology studies. In addition, the gene expression studies of Tox3 and Denndla in zebrafish demonstrated that N2 could impair PCOS condition. Furthermore, to confirm the N2 activity, the in‐silico studies were performed against PCOS susceptible genes Tox3 and Dennd1a using molecular docking and molecular dynamic simulations. The results suggested that N2 alleviated the oxidative stress and apoptosis in‐vitro and in‐vivo and altered the expression of PCOS key genes.