摘要目的 微RNA(miR)-21-5p对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞增殖、凋亡的影响及其分子作用机制.方法 建立大鼠PCOS模型,收集卵巢颗粒细胞,采用RT-qPCR检测PCOS模型大鼠和正常大鼠卵巢颗粒细胞中miR-21-5p和PDCD4mRNA表达,免疫印迹检测PDCD4蛋白表达;双荧光素酶报告基因实验检测miR-21-5p是否靶向PDCD4;将NC-miRNA、miR-21-5p-mimics、NC-siRNA、PDCD4-siRNA、pcDNA、pcDNA-PDCD4转染至PCOS大鼠卵巢颗粒细胞,MTT实验和流式细胞仪检测细胞增殖、凋亡情况.结果 与正常大鼠比较,PCOS大鼠卵巢颗粒细胞中miR-21-5p表达明显降低(P＜0.05),PDCD4 mRNA和蛋白表达明显增多(P＜0.05);双荧光素酶报告基因实验证实miR-21-5p靶向负调控PDCD4的表达;过表达miR-21-5p和抑制PDCD4表达均能促进PCOS大鼠卵巢颗粒细胞增殖并抑制细胞凋亡,而PDCD4过表达则可逆转miR-21-5p过表达对PCOS大鼠卵巢颗粒细胞增殖和凋亡的影响.结论 miR-21-5p通过靶向抑制PDCD4的表达,抑制PCOS大鼠卵巢颗粒细胞凋亡,促进细胞存活.

abstractsObjective To investigate the effect of miR-21-5p on proliferation and apoptosis of ovarian granulosa cells in rats with polycystic ovary syndrome (PCOS) and its molecular mechanism.Methods Rat PCOS model was established to collect ovarian granulosa cells.RT-qPCR was used to detect the expression of miR-21-5p and PDCD4 mRNA in ovarian granulosa cells of PCOS model rats and normal rats,Western blotting was used to detect the PDCD4 protein expression,and dual luciferase reporter gene assay was used to detect whether miR-21-5p was targeting PDCD4.NC-miRNA,miR-21-5p-mimics,NC-siRNA,PDCD4-siRNA,pcDNA,pcDNA-PDCD4 were transfected into PCOS rat ovarian granulosa cells,and cell proliferation and apoptosis were detected by MTT assay and flow cytometry.Results Compared with normal rats,the expression of miR-21-5p was significantly decreased (P＜0.05),and the expression of PDCD4 mRNA and protein was significantly increased (P＜0.05) in ovarian granulosa cells of PCOS rats.Dual luciferase reporter gene assay confirmed that miR-21-5p targeted and negatively regulated the expression of PDCD4.Overexpression of miR-21-5p and inhibition of PDCD4 could promote the proliferation and inhibit apoptosis of ovarian granulosa cells in PCOS rats,while overexpression of PDCD4 reversed the effect of miR-21-5p overexpression on proliferation and apoptosis of ovarian granulosa cells in PCOS rats.Conclusions miR-21-Sp inhibits apoptosis and promoted cell viability of ovarian granulosa cells in PCOS rats by targeting inhibition of PDCD4 expression.