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微小 RNA-21 通过靶向 Toll 样受体 TLR8 调节多囊卵巢综合征(PCOS)颗粒细胞的细胞凋亡和细胞增殖。

MicroRNA-21 regulate the cell apoptosis and cell proliferation of polycystic ovary syndrome (PCOS) granulosa cells through target toll like receptor TLR8.

机构信息

Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine Shanghai Jiao Tong University, Shanghai, China.

Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, China.

出版信息

Bioengineered. 2021 Dec;12(1):5789-5796. doi: 10.1080/21655979.2021.1969193.

Abstract

Polycystic ovary syndrome (PCOS) is a complex reproductive endocrine disease characterized by polycystic ovary. The aim of the study was to assess microRNA-21 regulates granulosa cell apoptosis and proliferation in polycystic ovary syndrome through target toll-like receptor 8. Granulosa cells were collected from 30 PCOS patients and 30 normal patients with tubal or male factor infertility (control) during in vitro fertilization-Embryo Transfer (IVF-ET) and were flash frozen with liquid nitrogen for storage for subsequent use. PCOS diagnosis was based on the revised standards of the American Society of Reproductive Medicine (ASRM) and the Rotterdam criteria PCOS granulosa cells and control granulosa cells were cultured in DMEM/F12 medium containing 10% fetal bovine serum and 1% antibiotic. After this RT-PCR, Western blot assessment and Detection of apoptosis by flow cytometry were conducted. The results of qPCR showed that the mRNA and protein expression of TLR8 in PCOS granulosa cells were significantly increased compared with the normal group. The results of Western blot also showed that the expression of TLR8, IFN-γ, TNF-α and IL-12 gene protein in the transfected cells was significantly higher than that in the control cells. Here, we show that miR-21 and TLR8 significantly increased in PCOS granulosa cell as compared with normal granulosa cells, and miR-21 enhances the TLR8 mRNA translation and then promotes the IFN-γ, TNF-α, and IL-12 secretion. Our study demonstrates that miR-21/ TLR8 involved in the PCOS inflammation, it provides profound insights into pathogenesis of PCOS.

摘要

多囊卵巢综合征(PCOS)是一种复杂的生殖内分泌疾病,其特征是多囊卵巢。本研究旨在评估 microRNA-21 通过靶向 toll 样受体 8 调节多囊卵巢综合征中颗粒细胞的凋亡和增殖。在体外受精-胚胎移植(IVF-ET)期间,从 30 名 PCOS 患者和 30 名因输卵管或男性因素不孕的正常患者(对照组)中收集颗粒细胞,并立即用液氮冷冻保存以备后用。PCOS 的诊断基于美国生殖医学协会(ASRM)的修订标准和 Rotterdam 标准 PCOS 颗粒细胞和对照颗粒细胞在含有 10%胎牛血清和 1%抗生素的 DMEM/F12 培养基中培养。在此之后进行 RT-PCR、Western blot 评估和流式细胞术检测细胞凋亡。qPCR 的结果表明,与正常组相比,PCOS 颗粒细胞中 TLR8 的 mRNA 和蛋白表达显著增加。Western blot 的结果也表明,转染细胞中 TLR8、IFN-γ、TNF-α 和 IL-12 基因蛋白的表达明显高于对照细胞。在这里,我们表明与正常颗粒细胞相比,miR-21 和 TLR8 在 PCOS 颗粒细胞中显著增加,并且 miR-21 增强了 TLR8 mRNA 的翻译,从而促进了 IFN-γ、TNF-α 和 IL-12 的分泌。我们的研究表明 miR-21/TLR8 参与了 PCOS 的炎症反应,为 PCOS 的发病机制提供了深刻的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d4a8/8806582/e961abc5ec90/KBIE_A_1969193_F0001_B.jpg

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