Kruppel like factor 12 gene regulates ovarian granulosa cells proliferation and apoptosis via the PI3K/Akt signaling pathway

Granulosa cells (GCs), as the largest proportion of somatic cells in mammalian ovarian follicles, play a central regulatory role in follicular development and oocyte maturation. In-depth analysis of the molecular mechanisms related to the growth and development and functional regulation of GCs is of great significance for improving the regulatory network mechanism in the field of female reproduction. Kruppel Like Factor 12 (KLF12) gene is involved in the regulation of various cellular physiological activities, but its specific regulatory function in ovarian GCs is not yet clear. In this study, the coding sequence (CDS) of KLF12 gene in New Zealand female rabbits was cloned for the first time, and the expression abundance and cellular localization of this gene in ovarian tissues of female rabbits at different reproductive stages were systematically analyzed. The results showed that KLF12 was highly expressed in the ovaries of 18-month-old female rabbits. Functional studies found that overexpression of KLF12 significantly inhibited the synthesis of Estradiol (E2) and Progesterone (P) in GCs, and the expression of genes related to cell proliferation, apoptosis and cycle development also changed. Further Western blot (WB) detection confirmed that KLF12 overexpression significantly reduced the phosphorylation levels of key molecules PI3K and Akt in the PI3K/Akt signaling pathway, while KLF12 knockdown had the opposite effect on GCs function. These results reveal the molecular mechanism of KLF12 regulating the development and function of ovarian GCs in New Zealand female rabbits through PI3K/Akt signaling pathway, which not only provides a theoretical basis for improving the reproductive performance of female rabbits, but also provides a new scientific perspective for the study of the molecular mechanism of mammalian reproductive regulation.