Characterization of the expression profile of FOinsulinXO3a and receptor (InsR) signaling pathway components in ovarian cells before and after treatment with 17²-estradiol and metformin

• Grant number: 19/27198-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2020
• End date: September 30, 2021
• Field of knowledge: Health Sciences - Medicine - Maternal and Child Health
• Principal Investigator: Edmund Chada Baracat
• Grantee: Lívia Marques Medeiros
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Estrogens account for numerous biological processes. In the ovaries, its production and secretion contribute significantly to the maintenance of female physiology. 17 ²-estradiol (E2) is the most active form of natural estrogen. To perform its function it depends on the activation of its receptor (ERa or b). In addition, it controls the PI3K / AKT signaling pathway by increasing its interaction with ERa, stimulating glucose metabolism. The antidiabetogenic potential of E2 may be of clinical importance in the treatment of insulin resistance (IR), commonly observed in polycystic ovary syndrome (PCOS). Although there has been a great advance in the understanding and characterization of energy change in IR, the number of drugs used in the treatment of IR-related morbidities is still very limited. The treatment of IR through insulin sensitizing agents, such as metformin, is still controversial, as some researchers advocate its use only in patients with glucose intolerance. Additionally, as already mentioned, E2 controls the PI3K / AKT pathway which, in response to insulin, induces FOXO transcription factor phosphorylation. This, in turn, can be inhibited by metformin, preventing protein translocation to the cytoplasm. Thus, there would be decreased expression of their target genes, which could induce IR. This work is part of a larger project aimed at evaluating the effects of 17²-estradiol and metformin treatment, isolated and combined, on glucose uptake and their role in FOXO3a and insulin receptor (InsR) signaling pathways in ovarian cells. in vitro. For this, initially, Hamster ovary CHO 1-15 (ATCC CRL9606) cells will be characterized for their expression profile of the components of the InsR and FOXO3A pathway by real time PCR and Western Blot. The expression of these markers will also be evaluated after treatment of cells with low and high glucose concentration (LowGlic = 1,000 mg / L and HighGlic = 1,800 mg / L, respectively).4These two groups will be subdivided into 8 new groups that will receive: 1) Control (vehicle only), 2) 17²-estradiol, 3) metformin, 4) insulin, 5) 17²-estradiol and metformin, 6) 17²-estradiol and insulin, 7) metformin and insulin; and 8) 17²-estradiol, metformin and insulin. The effect of treatments on protein expression of markers will be evaluated Western Blot. All results will be submitted to the relevant statistical analyzes.