中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (1): 10-20.doi: 10.12307/2026.539

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

人骨髓间充质干细胞增龄衰老不依赖于内源性反转录病毒的介导

王亚萍1，高天芸2，王  斌1，2   

  1. 1临床干细胞研究室，南京鼓楼医院，南京中医药大学鼓楼临床医学院，江苏省南京市  210008；2临床干细胞研究室，南京大学医学院附属鼓楼医院，江苏省南京市  210008
  • 收稿日期:2024-12-07 接受日期:2025-01-25 出版日期:2026-01-08 发布日期:2025-06-12
  • 通讯作者: 王斌，博士，教授，研究员，临床干细胞研究室，南京鼓楼医院，南京中医药大学鼓楼临床医学院，南京大学医学院附属鼓楼医院，江苏省南京市 210008
  • 作者简介:王亚萍，女，2001年生，湖北省十堰市人，汉族，南京中医药大学在读硕士，主要从事间充质干细胞临床应用的研究。
  • 基金资助:
    国家自然科学基金面上项目(82070459，82270701)，项目负责人：王斌

Senescence of human bone marrow mesenchymal stromal cells with increasing age is not dependent on the mediation of endogenous retroviruses

Wang Yaping1, Gao Tianyun2, Wang Bin1, 2   

  1. 1Clinical Stem Cell Center, Nanjing Drum Tower Hospital, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing 210008, Jiangsu Province, China; 2Clinical Stem Cell Center, Nanjing Drum Tower Hospital, Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China
  • Received:2024-12-07 Accepted:2025-01-25 Online:2026-01-08 Published:2025-06-12
  • Contact: Wang Bin, MD, Professor, Researcher, Clinical Stem Cell Center, Nanjing Drum Tower Hospital, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing 210008, Jiangsu Province, China; Clinical Stem Cell Center, Nanjing Drum Tower Hospital, Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, Jiangsu Province, China
  • About author:Wang Yaping, Master candidate, Clinical Stem Cell Center, Nanjing Drum Tower Hospital, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing 210008, Jiangsu Province, China
  • Supported by:
    National Natural Science Foundation of China (General Program), No. 82070459 and No. 82270701 (to WB)

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摘要：

文题释义：

骨髓间充质干细胞：为骨髓基质成纤维细胞，是一类起源于中胚层的成体干细胞，具有自我更新及多向分化潜能，可分化为多种间质组织，如骨髓、软骨、脂肪、骨髓造血组织等。此外，还具有独特的细胞因子分泌功能，使骨髓间充质干细胞在多种疾病中发挥一定的治疗效果。
内源性反转录病毒：是几百万年前古老的反转录病毒入侵人类基因组并整合的遗迹，约占人类基因组序列的8%，绝大多数的内源性反转录病毒会积累各种突变，从而在宿主基因组中失活或片段化，异常激活后会诱导疾病的发生，也会增加病毒蛋白的翻译以及病毒样颗粒在衰老细胞中的累积。

摘要
背景：人体增龄衰老可能是由于体内各种干细胞的衰老和耗竭。骨髓间充质干细胞具有重要的生理功能并对多种疾病展现出一定的疗效，研究骨髓间充质干细胞的衰老及机制具有重要意义。
目的：探讨人骨髓间充质干细胞是否伴随供者年龄的增加而呈现衰老表型，并进一步检测内源性反转录病毒是否驱动骨髓间充质干细胞的增龄衰老，为干细胞衰老机制研究提供新参考。 
方法：采用流式细胞术、β-半乳糖苷酶染色、qPCR、免疫印迹法及EdU荧光成像对不同年龄段的骨髓间充质干细胞进行衰老表征研究。收集不同年龄段供者的骨髓间充质干细胞及细胞培养上清，通过qPCR法检测人内源性反转录病毒的含量，进一步采用高灵敏的液滴式数字PCR检测细胞培养上清中内源性反转录病毒颗粒的表达，并用ELISA检测不同年龄段骨髓血浆样本中的内源性反转录病毒含量。
结果与结论：骨髓间充质干细胞具有明显的增龄衰老，包括显著的形态学改变、β-半乳糖苷酶染色阳性细胞比例升高、衰老标志物P16和P21蛋白表达升高，LMNB1蛋白表达下降，细胞增殖能力减弱。不同年龄段骨髓间充质干细胞中的内源性反转录病毒含量无显著性差异，培养上清中几乎不存在内源性反转录病毒颗粒，不同年龄段骨髓血浆样本中内源性反转录病毒无显著性差异。结果表明，人骨髓间充质干细胞具有正常生理性增龄衰老，但不是通过内源性反转录病毒的异常激活介导的，可能具有更加复杂的驱动机制。

关键词: 骨髓间充质干细胞, 衰老机制, 细胞衰老, 骨髓血浆, 内源性反转录病毒, 内源性反转录病毒颗粒, 液滴式数字PCR

Abstract: BACKGROUND: Aging of human body may be due to the senescence and depletion of various stem cells in the body. Bone marrow mesenchymal stromal cells have important physiological functions and have shown certain therapeutic effects on various diseases. It is of great significance to study the senescence and mechanism of bone marrow mesenchymal stromal cells. 
OBJECTIVE: To investigate whether human bone marrow mesenchymal stromal cells exhibit senescence phenotypes with increasing donor age, and further determine whether endogenous retrovirus drives the senescence of bone marrow mesenchymal stromal cells, offering a novel reference for the investigation of stem cell senescence mechanism.
METHODS: The senescence of bone marrow mesenchymal stromal cells at different ages was characterized by flow cytometry, β-galactosidase staining, qPCR, western blotting, and EdU fluorescence imaging. Bone marrow mesenchymal stromal cells and cell culture supernatant were collected from donors of different ages. The content of human endogenous retrovirus was detected by qPCR. Furthermore, highly sensitive droplet digital PCR was used to detect the expression of endogenous retrovirus-like particles in the cell culture supernatant. The content of endogenous retrovirus in bone marrow plasma samples of different ages was detected by ELISA.
RESULTS AND CONCLUSION: Bone marrow mesenchymal stromal cells exhibited obvious senescence with increasing age, including significant morphological changes, increased proportion of β-galactosidase positive cells, increased expression of senescence markers P16 and P21 protein, decreased expression of LMNB1 protein, and reduced cell proliferation ability. There was no significant difference in the content of endogenous retrovirus in bone marrow mesenchymal stromal cells at different ages, almost no endogenous retrovirus-like particles in the cell culture supernatant. There was no significant difference in endogenous retrovirus-like particles detected in bone marrow plasma samples at different ages. These findings indicate that human bone marrow mesenchymal stromal cells have normal physiological senescence with increasing age, but the mechanism of senescence is not mediated by abnormal activation of endogenous retroviruses, which may have a more complex driving mechanism. 

Key words: bone marrow mesenchymal stromal cell, aging mechanism, cellular senescence, bone marrow plasma, endogenous retrovirus, endogenous retrovirus-like particle, droplet digital PCR

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