Effects of miR-96 on the apoptosis and proliferation of Bos grunniens granulosa cells
The granulosa cells collected from adult Bos grunniens ovaries are taken as research mate-rials to investigate the effects of miR-96 on the apoptosis,proliferation and hormone secretion of the cells in Bos grunniens ovarian tissue.The cells are divided into four groups after isolation and cultiva-tion,transfected with miR-96 mimics and inhibitors as well as their negative controls(NC).The effects on the cells are cultured and detected as well.Flow cytometry and CCK-8 assay are used to detect cell apoptosis and proliferation.Besides,qPCR and Western Blot are used to detect the expres-sion of apoptosis-related genes.The steroid hormone secretion after transfection is detected by using ELISA.The results show that miR-96-mimics group inhibit granulosa cells apoptosis and promote cell proliferation.In addition,it significantly increases the expression of apoptosis-related gene and significantly decreases the expression of Bax.According to the variations steroid hormone levels de-tected by ELISA,it can be concluded that compared with mimics NC group,the secretion level of P4 in the cell culture medium of miR-96-mimics group is significantly reduced,while the secretion level of E2 is significantly increased.Compared with inhibitor group,the secretion level of P4 of miR-96-inhibitor group is significantly rised,while that of E2 is significantly decreased.In summa-ry,miR-96 can promote granulosa cell proliferation,inhibit apoptosis and affect reproductive hor-mone secretion,thereby participating in the regulation of follicular development and occlusion of Bos grunniens.
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