[1]张文豪,方锐,向文远,等.hUMSCs源性外泌体介导SIRT3/FOXO3通路改善软骨细胞衰老实验研究[J].陕西医学杂志,2025,54(6):723-728,735.[doi:DOI:10.3969/j.issn.1000-7377.2025.06.001]
　ZHANG Wenhao,FANG Rui,XIANG Wenyuan,et al.Humscs-derived exosomes mediate SIRT3/FOXO3 pathway to improve chondrocyte senescence[J].,2025,54(6):723-728,735.[doi:DOI:10.3969/j.issn.1000-7377.2025.06.001]
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hUMSCs源性外泌体介导SIRT3/FOXO3通路改善软骨细胞衰老实验研究

《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年6期
页码:
723-728,735
栏目:
基础研究
出版日期:
2025-06-05

文章信息/Info

Title:
Humscs-derived exosomes mediate SIRT3/FOXO3 pathway to improve chondrocyte senescence
作者:
张文豪1; 方锐1; 2; 3; 向文远1; 2; 3; 梁志权1; 2; 3
（1.新疆医科大学第四临床医学院，新疆 乌鲁木齐 830000；2.新疆医科大学附属中医医院，新疆 乌鲁木齐 830000;3.新疆维吾尔自治区中医药研究院，新疆 乌鲁木齐 830000）
Author(s):
ZHANG Wenhao1; FANG Rui1; 2; 3; XIANG Wenyuan1; 2; 3; LIANG Zhiquan1; 2; 3
(1.The Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China;2.The Affiliated Hospital of Traditional Chinese Medicine of Xinjiang Medical University,Urumqi 830000,China;3.Academy of Traditional Chinese Medicine，Xinjiang Uygur Autonomous Region，Urumqi 830000, China)
关键词:
骨关节炎; 脐带间充质干细胞; 外泌体; 沉默信息调节因子3/叉头框蛋白O3通路; 衰老; 软骨细胞
Keywords:
Osteoarthritis; Umbilical cord mesenchymal stem cells; Exosomes; SIRT3/FOXO3 pathway; Senescence; Chondrocytes
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2025.06.001
文献标志码:
A
摘要:
目的：探讨人脐带间充质干细胞（hUMSCs）来源的外泌体（hUMSCs-Exos）对软骨细胞衰老的影响。方法：培养hUMSCs后提取hUMSCs-Exos，根据浓度分为高浓度组（60 μg/ml）、中浓度组（40 μg/ml）和低浓度组（20 μg/ml），并设置空白组（0 μg/ml）。采用透射电子显微镜（TEM）和纳米粒子跟踪分析（NTA）对hUMSCs-Exos进行形态和尺寸分布鉴定。CCK-8法检测第3、5、7、9、13天的细胞增殖情况。使用过氧化氢处理软骨细胞构建衰老模型（模型组）。对衰老模型软骨细胞进行衰老相关β-半乳糖苷酶（SA-β-Gal）染色，统计阳性细胞数量。Western blot和实时定量PCR（RT-qPCR）检测沉默信息调节因子3（SIRT3）/叉头框蛋白O3（FOXO3）通路SIRT3、FOXO3、Bcl-2相关X蛋白（Bax）蛋白及mRNA表达水平。结果：TEM显示hUMSCs-Exos呈杯状或半球形，NTA分析表明其直径主要分布在60～200 nm，符合外泌体特征。在第3、5、7、9、13天，与空白组比较，低浓度组和中浓度组软骨细胞增殖活性升高，高浓度组软骨细胞增殖活性降低(均P<0.05)；与低浓度组比较，中浓度组软骨细胞增殖活性升高，高浓度组软骨细胞增殖活性降低(均P<0.05)；与中浓度组比较，高浓度组软骨细胞增殖活性降低(P<0.05)。SA-β-Gal染色结果显示，模型组软骨细胞衰老百分比较空白组增加，中浓度组软骨细胞衰老百分比较模型组降低(均P<0.05)。Western blot和RT-qPCR结果显示，与空白组比较，模型组软骨细胞SIRT3、FOXO3蛋白和mRNA表达降低，Bax蛋白和mRNA表达升高(均P<0.05)；与模型组比较，低浓度组与中浓度组软骨细胞SIRT3、FOXO3蛋白和mRNA表达升高，Bax蛋白和mRNA表达降低(均P<0.05)；与低浓度组比较，中浓度组软骨细胞SIRT3、FOXO3蛋白和mRNA表达降低，Bax蛋白和mRNA表达升高(均P<0.05)。结论：hUMSCs-Exos能够显著促进软骨细胞增殖并抑制其衰老过程，其作用机制可能与上调SIRT3/FOXO3通路表达有关。
Abstract:
Objective:To investigate the effects of human umbilical cord mesenchymal stem cells (hUMSCs)-derived exosomes (HUMSCs-Exos) on the senescence of chondrocytes.Methods：After culturing hUMSCs,hUMSCs-Exos were extracted and divided into high concentration group (60 μg/ml),medium concentration group (40 μg/ml),low concentration group (20 μg/ml),and a blank group (0 μg/ml).The morphology and size distribution of hUMSCs-Exos were identified by using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA).The CCK-8 assay was used to assess cell proliferation on days 3,5,7,9 and 13.Chondrocytes were treated with hydrogen peroxide to establish an aging model (model group).Senescence-associated β-galactosidase (SA-β-Gal) staining was performed on aging model chondrocytes to quantify the number of positive cells.Western blot and RT-qPCR were used to analyze the expression levels of silent information regulator 3 (SIRT3) and forkhead box protein O3 (FOXO3) pathway-related SIRT3,FOXO3 and Bax proteins and mRNAs.Results:TEM showed that hUMSCs-Exos were cup-shaped or spherical,and NTA analysis indicated that the diameter was primarily distributed between 60 to 200 nm,consistent with the characteristics of exosomes.On days 3,5,7,9 and 13,compared with the blank group,the proliferation activity of chondrocytes in the low concentration group and the medium concentration group was increased,and the proliferation activity of chondrocytes in the high concentration group was decreased (all P<0.05);compared with the low concentration group,the proliferation activity of chondrocytes in the medium concentration group was increased,and the proliferation activity of chondrocytes in the high concentration group was decreased (both P<0.05);compared with the medium concentration group,the proliferation activity of chondrocytes in the high concentration group was decreased (P<0.05).The results of SA-β-Gal staining revealed that the percentage of aging chondrocytes in the model group significantly increased compared to the blank control group,while the percentage in the medium concentration group decreased significantly compared to the model group (all P<0.05).Western blot and RT-qPCR results indicated that compared with the blank group,SIRT3 and FOXO3 protein and mRNA expressions were significantly reduced,whereas Bax protein and mRNA expressions were significantly increased in the model group (all P<0.05);compared with the model group,both the medium and low concentration groups showed elevated SIRT3 and FOXO3 protein and mRNA expressions,with reduced Bax protein and mRNA expressions (all P<0.05);compared with the low concentration group,SIRT3 and FOXO3 protein and mRNA expressions in the medium concentration group were lower,whereas Bax protein and mRNA expressions were higher (all P<0.05).Conclusion:The hUMSCs-Exos can significantly promote chondrocyte proliferation and inhibit their aging process,which may be related to the up-regulation of SIRT3/FOXO3 pathway.

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备注/Memo

备注/Memo:
国家自然科学基金资助项目（82405423，82360934）；新疆维吾尔自治区自然科学基金资助项目（2023D01C145）；新疆维吾尔自治区重点研发计划项目（2021B03006）；新疆维吾尔自治区科技创新领军人才项目（2022TSYCLJ0007）；新疆维吾尔自治区青年托举人才项目（2023TSYCQNTJ0050）
更新日期/Last Update: 2025-06-04