Adipose-derived stem cells exosomal KLF3-AS1 attenuates ovarian function by YBX1/PI3K/Akt/mTOR signaling.

IF 2.2 4区医学 Q3 PHYSIOLOGY

Physiology international Pub Date : 2025-04-10 DOI:10.1556/2060.2025.00357

Wei Zhao, Haili Zhang, Liyan Zhang, Caizhu Hai, Shujun Liu, Haiyan Li, Yanan Zhang, Hongwu Wang, Caisheng Wang

{"title":"Adipose-derived stem cells exosomal KLF3-AS1 attenuates ovarian function by YBX1/PI3K/Akt/mTOR signaling.","authors":"Wei Zhao, Haili Zhang, Liyan Zhang, Caizhu Hai, Shujun Liu, Haiyan Li, Yanan Zhang, Hongwu Wang, Caisheng Wang","doi":"10.1556/2060.2025.00357","DOIUrl":null,"url":null,"abstract":"Background: Adipose-derived stem cell (ADSC) derived exosomes have been widely studied in disease treatment. Exosomes are able to deliver bioactive molecules, including non-coding RNAs and proteins. Long non-coding RNAs (lncRNAs) are non-coding RNAs longer than 200 nucleotides and are enriched in exosomes. This work aimed to explore the effects of lncRNA KLF3 antisense RNA 1 (KLF3-AS1) that delivered by ADSC-derived exosomes on ovarian aging.Methods: ADSCs were isolated and characterized with the surface biomarkers. Exosomes were isolated from ADSCs. The biomarkers of ADSC-derived exosomes were identified using western blotting. Exosomes were labeled with PKH26 and internalized by primary granulosa cells (pGCs), and relative images were taken under fluorescence microscope. ADSCs were transfected with KLF3-AS1, and exosomes were isolated for treatment of aging female mice. The ovary weight was recorded. The follicular development was measured by Hematoxylin and eosin (H&E) staining and Masson's trichrome staining. Apoptosis of ovary tissues was detected by TUNEL assay. The senescence and apoptosis of pGCs were determined by S-β-gal staining kit and Annexin V/PI detection kit. RNA pulldown and RNA Immunoprecipitation Chip (RIP) assay were performed to determine the interaction of Y box binding protein 1 (YBX1) with KLF3-AS1.Results: The ADSC-derived exosomes could deliver KLF3-AS1 to pGCs. Treatment with ADSC-derived exosomes notably elevated the ovary weight and enhanced follicular development in aged mice, whereas depletion of KLF3-AS1 reversed these effects and promoted cell apoptosis. ADSCs-derived exosomes alleviated senescence and apoptosis of pGCs, while KLF3-AS1 depletion blocked these phenotypes. KLF3-AS1 directly interacts with YBX1. KLF3-AS1 depletion inhibited phosphorylation of PI3K, Akt, and mTOR in pGC, and overexpression of YBX1 reversed these phenotypes.Conclusion: ADSC-derived exosomal KLF3-AS1 could improve ovary aging and enhance pGC viability via targeting the YBX1 and PI3K/AKT/mTOR signaling.","PeriodicalId":20058,"journal":{"name":"Physiology international","volume":" ","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Physiology international","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1556/2060.2025.00357","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"PHYSIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Background: Adipose-derived stem cell (ADSC) derived exosomes have been widely studied in disease treatment. Exosomes are able to deliver bioactive molecules, including non-coding RNAs and proteins. Long non-coding RNAs (lncRNAs) are non-coding RNAs longer than 200 nucleotides and are enriched in exosomes. This work aimed to explore the effects of lncRNA KLF3 antisense RNA 1 (KLF3-AS1) that delivered by ADSC-derived exosomes on ovarian aging.

Methods: ADSCs were isolated and characterized with the surface biomarkers. Exosomes were isolated from ADSCs. The biomarkers of ADSC-derived exosomes were identified using western blotting. Exosomes were labeled with PKH26 and internalized by primary granulosa cells (pGCs), and relative images were taken under fluorescence microscope. ADSCs were transfected with KLF3-AS1, and exosomes were isolated for treatment of aging female mice. The ovary weight was recorded. The follicular development was measured by Hematoxylin and eosin (H&E) staining and Masson's trichrome staining. Apoptosis of ovary tissues was detected by TUNEL assay. The senescence and apoptosis of pGCs were determined by S-β-gal staining kit and Annexin V/PI detection kit. RNA pulldown and RNA Immunoprecipitation Chip (RIP) assay were performed to determine the interaction of Y box binding protein 1 (YBX1) with KLF3-AS1.

Results: The ADSC-derived exosomes could deliver KLF3-AS1 to pGCs. Treatment with ADSC-derived exosomes notably elevated the ovary weight and enhanced follicular development in aged mice, whereas depletion of KLF3-AS1 reversed these effects and promoted cell apoptosis. ADSCs-derived exosomes alleviated senescence and apoptosis of pGCs, while KLF3-AS1 depletion blocked these phenotypes. KLF3-AS1 directly interacts with YBX1. KLF3-AS1 depletion inhibited phosphorylation of PI3K, Akt, and mTOR in pGC, and overexpression of YBX1 reversed these phenotypes.

Conclusion: ADSC-derived exosomal KLF3-AS1 could improve ovary aging and enhance pGC viability via targeting the YBX1 and PI3K/AKT/mTOR signaling.

查看原文本刊更多论文

脂肪源性干细胞外泌体KLF3-AS1通过YBX1/PI3K/Akt/mTOR信号通路减弱卵巢功能。

背景：脂肪源性干细胞（ADSC）衍生外泌体在疾病治疗中得到了广泛的研究。外泌体能够传递生物活性分子，包括非编码rna和蛋白质。长链非编码rna （lncRNAs）是长度超过200个核苷酸的非编码rna，富集于外泌体中。本研究旨在探讨由adsc来源的外泌体递送的lncRNA KLF3反义RNA 1 （KLF3- as1）对卵巢衰老的影响。方法：分离ADSCs，用表面生物标志物对其进行鉴定。从ADSCs中分离外泌体。adsc衍生外泌体的生物标志物采用western blotting鉴定。外泌体用PKH26标记，由原代颗粒细胞（pGCs）内化，荧光显微镜下拍摄相关图像。用KLF3-AS1转染ADSCs，分离外泌体治疗衰老雌性小鼠。记录卵巢重量。采用苏木精-伊红（H&E）染色和马松三色染色检测卵泡发育。TUNEL法检测卵巢组织凋亡。采用S-β-gal染色试剂盒和Annexin V/PI检测试剂盒检测pGCs的衰老和凋亡情况。采用RNA pull - down和RNA Immunoprecipitation Chip （RIP）检测Y box binding protein 1 （YBX1）与KLF3-AS1的相互作用。结果：adsc衍生的外泌体可以向pGCs传递KLF3-AS1。adsc来源的外泌体显著提高了老年小鼠的卵巢重量和卵泡发育，而KLF3-AS1的缺失逆转了这些作用并促进了细胞凋亡。adscs来源的外泌体减轻了pGCs的衰老和凋亡，而KLF3-AS1缺失阻断了这些表型。KLF3-AS1直接与YBX1相互作用。KLF3-AS1缺失抑制pGC中PI3K、Akt和mTOR的磷酸化，而YBX1的过表达逆转了这些表型。结论：adsc来源的外泌体KLF3-AS1可通过靶向YBX1和PI3K/AKT/mTOR信号通路改善卵巢衰老，提高pGC活力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Physiology international Medicine-Physiology (medical)

CiteScore

3.40

自引率

0.00%

发文量

期刊介绍： The journal provides a forum for important new research papers written by eminent scientists on experimental medical sciences. Papers reporting on both original work and review articles in the fields of basic and clinical physiology, pathophysiology (from the subcellular organization level up to the oranizmic one), as well as related disciplines, including history of physiological sciences, are accepted.