脐带间充质干细胞来源的细胞外囊泡通过METTL3改善颗粒细胞过度自噬。

IF 5 2区 生物学 Q2 CELL BIOLOGY
Weiqin Zhou, Ju Zhang, Xuanping Lu, Ziwei Zhao, Yujing Weng, Chunrong Zhu
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引用次数: 0

摘要

目的:多囊卵巢综合征(PCOS)是一种影响女性生育能力的常见内分泌疾病。我们通过甲基转移酶样3 (METTL3)评估了脐带间充质干细胞衍生的细胞外囊泡(UC-MSC-EVs)对pten诱导的激酶1 (PINK1)/帕金森介导的卵巢颗粒细胞(GCs)过度自噬的影响。方法:培养人卵巢GC细胞系KGN,用脱氢表雄酮(DHEA)和uc - msc - ev处理。通过流式细胞术、CCK-8、western blot、kit和免疫荧光检测细胞凋亡和活力、自噬相关蛋白水平、三磷酸腺苷(ATP)和线粒体膜电位(MMP)水平、微管相关蛋白1轻链3 β (LC3B)和线粒体外膜转位酶20 (TOMM20)共定位。采用Me-RIP、RT-qPCR和western blot检测PINK1 n6 -甲基腺苷(m6A)修饰、METTL3水平和PINK1 mRNA稳定性。建立PCOS小鼠模型并给予uc - msc - ev治疗。ELISA法检测血清激素及卵巢组织自噬相关蛋白水平。结果:DHEA降低KGN细胞活力和p62水平,增加PINK1、Parkin、LC3BII/I和Beclin-1蛋白水平、ATP含量、MMP水平、TOMM20+LC3B+细胞数量和凋亡,而uc - msc - ev处理部分消除了这些作用。PINK1有m6A修饰位点。METTL3是PINK1 m6a修饰的Writer蛋白。DHEA处理后,KGN细胞METTL3和PINK1 m6A修饰水平升高,mRNA稳定性提高,而UC-MSC-EV处理的结果相反。METTL3过表达部分避免了uc - msc - ev改善的PINK1/ parkin介导的有丝分裂。uc - msc - ev通过METTL3抑制PINK1/ parkin介导的过度自噬,改善PCOS小鼠卵巢功能。结论:uc - msc - ev通过METTL3抑制PINK1/ parkin介导的卵巢GCs的有丝分裂,从而改善PCOS。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Umbilical cord mesenchymal stem cells-derived extracellular vesicles improve excessive autophagy of granulosa cells through METTL3.

Objective: Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder impacting women's fertility. We assessed the effect of umbilical cord mesenchymal stem cell-derived extracellular vesicles (UC-MSC-EVs) on PTEN-induced kinase 1 (PINK1)/Parkin-mediated excessive autophagy of ovarian granulosa cells (GCs) through methyltransferase-like 3 (METTL3). Methods: Human ovarian GC line KGN was cultured and treated with dehydroepiandrosterone (DHEA) and UC-MSC-EVs. Cell apoptosis and viability, autophagy-related protein levels, adenosine triphosphate (ATP) and mitochondrial membrane potential (MMP) level, and microtubule-associated protein 1 light chain 3 beta (LC3B) and translocase of outer mitochondrial membrane 20 (TOMM20) co-localization were assessed by flow cytometry, CCK-8, western blot, kit, and immunofluorescence. PINK1 N6-methyladenosine (m6A) modification, METTL3 levels, and PINK1 mRNA stability were determined by Me-RIP, RT-qPCR and western blot. The PCOS mouse model was established and treated with UC-MSC-EVs. Serum hormone and ovarian tissue autophagy-related protein levels were determined by ELISA. Results: DHEA decreased KGN cell viability and p62 level, increased PINK1, Parkin, LC3BII/I and Beclin-1 protein levels, ATP content, MMP level, TOMM20+LC3B+ cell number and apoptosis, which were partly abrogated by UC-MSC-EVs treatment. PINK1 had m6A modification sites. METTL3 was a PINK1 m6A-modified Writer protein. After DHEA treatment, KGN cells showed elevated METTL3 and PINK1 m6A modification levels and mRNA stability, while UC-MSC-EV treatment caused the opposite results. METTL3 overexpression partly averted UC-MSC-EVs-improved PINK1/Parkin-mediated mitophagy. UC-MSC-EVs curbed PINK1/Parkin-mediated excessive autophagy through METTL3 and improved ovarian function in PCOS mice. Conclusions: UC-MSC-EVs suppressed PINK1/Parkin-mediated mitophagy of ovarian GCs through METTL3, thereby improving PCOS.

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来源期刊
CiteScore
9.10
自引率
1.80%
发文量
252
审稿时长
1 months
期刊介绍: The American Journal of Physiology-Cell Physiology is dedicated to innovative approaches to the study of cell and molecular physiology. Contributions that use cellular and molecular approaches to shed light on mechanisms of physiological control at higher levels of organization also appear regularly. Manuscripts dealing with the structure and function of cell membranes, contractile systems, cellular organelles, and membrane channels, transporters, and pumps are encouraged. Studies dealing with integrated regulation of cellular function, including mechanisms of signal transduction, development, gene expression, cell-to-cell interactions, and the cell physiology of pathophysiological states, are also eagerly sought. Interdisciplinary studies that apply the approaches of biochemistry, biophysics, molecular biology, morphology, and immunology to the determination of new principles in cell physiology are especially welcome.
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