Isatin-linked pyrazole K1 derivative alter the phosphatidylinositol-3-kinase pathway by enhancing the metabolic function and folliculogenesis in the triclosan-induced PCOS-like condition in zebrafish model

Abstract

Polycystic ovarian syndrome (PCOS), which causes hormonal imbalance, inflammation, and metabolic disorders, requires several treatments. This study aimed to examine the Isatin-linked pyrazole K1 derivative's effectiveness in PCOS induced by environmental contaminants such as triclosan, specifically assessing its biochemical, metabolic, and reproductive impacts. Isatin-linked pyrazole K1 derivative was synthesised in the lab and tested in vitro and in vivo, including cytotoxicity testing in CHO cells, apoptosis analysis in AO/PI staining, and developmental toxicity in zebrafish embryos. In addition, for network pharmacology analysis, BindingDB, GeneCard, and other databases were used to characterise the interaction of K1 derivative with PCOS-related genes and pathways, followed by examining the apoptosis in CHO cells, estimation of total cholesterol and triglycerides in adipose tissue of zebrafish. Furthermore, GSI%, follicular stage examination, collagen accumulation, nucleic acid staining by toluidine blue, and gene expression of cyp19a1a, dennd1a, tox3, pik3ca, and pik3cd were examined. The research found that K1 reduces various PCOS pathologies, improving folliculogenesis, overall ovarian function, and follicular growth. K1 treatment at 25 µM significantly enhanced SOD (1.470 ± 0.01533 U/ml), CAT (1.174 ± 0.008687 U/ml), and GSH (1.375 ± 0.006409 U/ml) levels while reducing LDH activity (0.9815 ± 0.01273 nmol/mg), demonstrating its ability to mitigate oxidative stress and cellular damage. In particular, K1 modulates insulin sensitivity by reducing the blood glucose level in PCOS-induced fish and lowering lipid levels, which is essential for treating PCOS metabolic symptoms. K1 derivative also significantly reduced collagen deposition in ovarian tissues, indicating K1 may reduce PCOS-related fibrosis, which suggests that the derivative may be a novel therapeutic agent for PCOS. The comprehensive approach of K1 addresses metabolic and reproductive concerns; however, clinical studies must be conducted to test these findings' efficacy and safety and understand its therapeutic molecular processes.