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miR-130b regulates gap junctional intercellular communication through connexin 43 in granulosa cells from patients with polycystic ovary syndrome.

MiR-130b 通过缝隙连接蛋白 43 调节多囊卵巢综合征患者颗粒细胞的缝隙连接细胞间通讯。

  • 影响因子:3.06
  • DOI:10.1093/molehr/gaaa044
  • 作者列表:"Jiang L","Huang H","Qian Y","Li Y","Chen X","Di N","Yang D
  • 发表时间:2020-06-19
Abstract

: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. 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 gap junctional intercellular communication 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 gap junctional intercellular communication. 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.

摘要

: MicroRNAs (miRNAs) 是一种小的非编码 rna,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) 妇女的黄素化颗粒细胞中的表达是否与正常月经周期妇女的黄素化颗粒细胞不同。以及某些 miRNAs 是否调节 Cx43 水平和缝隙连接细胞间通讯。使用 miRNA 微阵列研究了 5 例 PCOS 女性和 5 例无 PCOS 女性的卵巢皮质组织中的 miRNA 谱。采用实时荧光定量 PCR (real-time PCR) 检测 20 例 PCOS 患者和 25 例非 PCOS 患者卵巢黄素化颗粒细胞的 miR-130b 和 Cx43 mRNA 水平。进行蛋白和 mRNA 表达分析和荧光素酶测定以确认 miR-130b 的底物。PCOS 卵巢皮质与非 PCOS 卵巢皮质相比,miRNAs 的表达存在差异。此外,与正常月经周期的女性相比,PCOS 卵巢皮质和黄素化颗粒细胞中的 miR-130b 水平升高,而 Cx43 mRNA 水平,miR-130b 的确定靶点,PCOS 患者的颗粒细胞减少。颗粒细胞系中 miR-130b 的过表达导致 Cx43 蛋白水平降低,并使用刮负荷和染料转移试验抑制缝隙连接细胞间通讯。同时,抑制 miR-130b 增加了 Cx43 水平。总之,PCOS 颗粒细胞中的 miR-130b 增加,其靶向 Cx43 影响缝隙连接细胞间通讯。本研究结果提示 miR-130b 通过转录后调控 Cx43 参与 PCOS 的病理生理过程,为 PCOS 的发病机制提供了新的思路。

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影响因子:3.62
发表时间:2020-01-15
DOI:10.1016/j.mce.2019.110611
作者列表:["Makrinou E","Drong AW","Christopoulos G","Lerner A","Chapa-Chorda I","Karaderi T","Lavery S","Hardy K","Lindgren CM","Franks S"]

METHODS:Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder amongst women of reproductive age, whose aetiology remains unclear. To improve our understanding of the molecular mechanisms underlying the disease, we conducted a genome-wide DNA methylation profiling in granulosa lutein cells collected from 16 women suffering from PCOS, in comparison to 16 healthy controls. Samples were collected by follicular aspiration during routine egg collection for IVF treatment. Study groups were matched for age and BMI, did not suffer from other disease and were not taking confounding medication. Comparing women with polycystic versus normal ovarian morphology, after correcting for multiple comparisons, we identified 106 differentially methylated CpG sites with p-values <5.8 × 10 that were associated with 88 genes, several of which are known to relate either to PCOS or to ovarian function. Replication and validation of the experiment was done using pyrosequencing to analyse six of the identified differentially methylated sites. Pathway analysis indicated potential disruption in canonical pathways and gene networks that are, amongst other, associated with cancer, cardiogenesis, Hedgehog signalling and immune response. In conclusion, these novel findings indicate that women with PCOS display epigenetic changes in ovarian granulosa cells that may be associated with the heterogeneity of the disorder.

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影响因子:2.68
发表时间:2020-02-01
DOI:10.1080/14656566.2019.1701655
作者列表:["Vatopoulou A","Tziomalos K"]

METHODS::Introduction: Approximately 1% of adolescents have polycystic ovary syndrome (PCOS) and almost 40-70% of these patients are overweight or obese. Obese adolescents with PCOS have more severe insulin resistance and hyperandrogenemia, a more adverse lipid profile and a worse quality of life than normal-weight adolescents with PCOS. Accordingly, weight loss is an important component of the management of these patients.Areas covered: The authors discuss the different options for weight loss in obese adolescents with PCOS. Lifestyle changes appear to be effective but adherence to this intervention is suboptimal. There are also limited data regarding the optimal diet in this population. Few small studies have evaluated the effects of pharmacotherapy in these patients. Conflicting data have been reported regarding the effects of metformin on body weight. Notably, agents that have been approved for weight loss in adults have not been evaluated in adolescents with PCOS.Expert opinion: More studies are needed to identify the most appropriate diet for obese adolescents with PCOS. Well-designed randomized controlled studies are also needed to define the safety and efficacy of pharmacotherapy in this population.

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影响因子:2.78
发表时间:2020-02-01
DOI:10.1007/s11010-019-03678-6
作者列表:["Ding Y","He P","Li Z"]

METHODS::Polycystic ovary syndrome (PCOS) is a hormonal disorder common among women of reproductive age. Although much is understood concerning the pathology of PCOS, further investigation into the influence of microribonucleic acids (miRNAs) on the proliferation of ovarian granulosa cells (GCs) is needed. This study investigated the role of specific miRNAs in ovarian dysfunction of PCOS and its effect on the proliferation of GCs. Initially, miRNA profiling was performed on the ovarian cortexes of 15 rats in which PCOS had been induced and 15 rats without PCOS (non-PCOS). This mechanical study was performed on ovarian GCs extracted from human chorionic gonadotrophin (hCG)-induced rats. Insulin was used to treat GCs to establish the PCOS cell model. Increased Equus caballus mir-9119 expression was observed and confirmed in the insulin-induced model of PCOS in GCs (GC-PCOS) as well as in the hCG-induced rats when compared to non-PCOS rats and cells. Observation and confirmation were carried out through both miRNA array and quantitative PCR. In contrast, downregulation of the nuclear factor kappa B (NFκB) p65 was observed in the PCOS cell model. Additionally, annexin V, FITC, and propidium iodide flow cytometry showed overexpression of miR-9119-induced apoptosis. In this study, we revealed that miR-9119 inhibition regulates p65 expression levels in insulin-treated GCs by binding to the 3'-untranslated of p65. Additionally, regulation of p65 expression was positively correlated with the expression of the double-stranded RNA endoribonuclease DICER. Moreover, RNA silencing/overexpression of p65 affected the functional role of miR-9119. In conclusion, GCs of PCOS, the expression of miR-9119, and targeted NFκB/p65-DICER axis are upregulated in order to maintain cell viability and prevent apoptosis, thereby promoting Anti-Müllerian hormone production in GCs. This study may provide a new understanding of the mechanism of GC dysfunction.

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