Metabolic identification of bioactive compounds of Citrus reticulata cultivars extracts for a novel approach to polycystic ovary syndrome.

Polycystic ovary syndrome; is a prevalent endocrine disorder, characterized by hormonal imbalances, inflammation, and oxidative stress. Natural remedies, including Murcott and Merav (Citrus reticulata varieties, Family: Rutaceae) fruits and leaves extracts, have demonstrated promising therapeutic effects in alleviating these symptoms. This study aimed to evaluate the efficacy of these extracts concerning their metabolomics in modulating hormonal levels, enhancing lipid metabolism while alleviating inflammation and oxidative stress in mice with polycystic ovary syndrome through the regulation of miR-33b-5P and miR-145 gene expression. UPLC-T-TOF-MS/MS analysis identified and quantified One hundred-seven bioactive metabolites. These metabolites were categorized into various classes according to their chemical structures, including phenolic types such as 68 flavonoids, 4 coumarins and 7 phenolic acids in addition to, 3 nitrogenous compounds, 7 terpenes and terpenoids, 6 fatty acids and their conjugates and 12 miscellaneous compounds. and various biochemical assays were performed. Blood samples were collected to measure plasma FSH, testosterone, LH, and lipid profiles. Oxidative stress biomarkers, including malondialdehyde (MDA) and superoxide dismutase (SOD), and COX-2 and NF-kB levels were assessed using ELISA kits. Micro-RNA gene expression, miR-33b-5P and miR-145, related to inflammation and oxidative stress, was analyzed using qRT-PCR. Ovarian tissues were examined histologically to assess morphological changes. These findings highlight that Murcott and Merav extracts could be a promising alternative natural treatment for PCOS, balancing hormonal levels, improving lipid profiles, and reducing inflammation and oxidative stress. Merav leaves emerged as the most promising candidate for PCOS therapy, attributed to their high levels of methylated kaempferol, a key bioactive flavonoid.