MicroRNAs (miRNAs) are small, noncoding RNAs that negatively regulate gene expression post-transcriptionally. We explored whether connexin 43 (Cx43) was differently expressed in luteinized granulosa cells from women with polycystic ovary syndrome (PCOS) compared with luteinized granulosa cells from women with a normal menstrual cycle, and whether certain miRNAs regulate the Cx43 level and gap junctional intercellular communication (GJIC). The miRNA profile was investigated in ovarian cortex tissues from five women with PCOS and five women without PCOS using a miRNA microarray. The levels of miR-130b and Cx43 mRNA were measured using real-time PCR in human luteinized granulosa cells from 20 women with PCOS and 25 women without PCOS. Protein and mRNA expression analysis and luciferase assays were conducted to confirm the substrate of miR-130b. PCOS ovarian cortex showed differential expression of miRNAs compared with non-PCOS ovarian cortex. Furthermore, miR-130b levels were increased in PCOS ovarian cortex and in luteinized granulosa cells compared with those in women with normal menstrual cycles, whereas the level of Cx43 mRNA, the identified target of miR-130b, was decreased in granulosa cells from patients with PCOS. Overexpression of miR-130b in a granulosa cell line resulted in reduced Cx43 protein levels and inhibited GJIC using scrape loading and dye transfer assay. Meanwhile, inhibition of miR-130b increased the Cx43 level. In conclusion, miR-130b was increased in PCOS granulosa cells, where it targets Cx43 to affect GJIC. The results of the present study suggested that miR-130b, via post-transcriptional regulation of Cx43, is involved in the pathophysiology of PCOS, which provides new insight into the pathological mechanism of PCOS.

中文翻译：

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miR-130b 通过多囊卵巢综合征患者颗粒细胞中的连接蛋白 43 调节间隙连接细胞间通讯。

MicroRNA (miRNA) 是小的非编码 RNA，可在转录后负调控基因表达。我们探讨了连接蛋白 43 (Cx43) 在多囊卵巢综合征 (PCOS) 女性的黄素化颗粒细胞中的表达是否与月经周期正常女性的黄素化颗粒细胞不同，以及某些 miRNA 是否调节 Cx43 水平和间隙连接细胞间通讯(GJIC)。使用 miRNA 微阵列在来自五名患有 PCOS 的女性和五名没有 PCOS 的女性的卵巢皮质组织中研究了 miRNA 谱。miR-130b 和 Cx43 mRNA 的水平使用实时 PCR 在来自 20 名 PCOS 女性和 25 名非 PCOS 女性的人黄素化颗粒细胞中测量。进行蛋白质和 mRNA 表达分析以及荧光素酶测定以确认 miR-130b 的底物。与非 PCOS 卵巢皮质相比，PCOS 卵巢皮质显示出 miRNA 的差异表达。此外，与月经周期正常的女性相比，PCOS 卵巢皮质和黄体化颗粒细胞中的 miR-130b 水平升高，而 miR-130b 的鉴定靶标 Cx43 mRNA 水平在患有以下疾病的患者的颗粒细胞中降低。多囊卵巢综合征。miR-130b 在颗粒细胞系中的过表达导致 Cx43 蛋白水平降低，并使用刮片加载和染料转移测定抑制 GJIC。同时，抑制 miR-130b 增加了 Cx43 水平。总之，miR-130b 在 PCOS 颗粒细胞中增加，它靶向 Cx43 以影响 GJIC。本研究结果表明 miR-130b 通过 Cx43 的转录后调控参与 PCOS 的病理生理，