装载有β-烟酰胺单核苷酸的间充质基质细胞衍生的小细胞外囊泡激活NAD/SIRT3信号通路介导的线粒体自噬以延缓皮肤衰老。

Small extracellular vesicles derived from mesenchymal stromal cells loaded with β-nicotinamide mononucleotide activate NAD/SIRT3 signaling pathway-mediated mitochondrial autophagy to delay skin aging.

作者信息

Sun Zixuan, Li Jiali, Zheng Yuzhou, Zhang Jiaxin, Bai Wenhuan, Deng Xinyi, Wu Zhijing, Xu XueZhong, Ding Wei, Qian Hui, Tan Yulin

机构信息

Department of General Surgery, Wujin Hospital Affiliated with Jiangsu University, Changzhou, 213017, China.

Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, Department of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, 212013, China.

出版信息

Stem Cell Res Ther. 2025 Jul 1;16(1):339. doi: 10.1186/s13287-025-04460-w.

DOI:10.1186/s13287-025-04460-w

PMID:40598314

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12218950/

Abstract

BACKGROUND

Recently, the beneficial effects of human umbilical cord mesenchymal stromal cell (hucMSC)-derived small extracellular vesicles (sEVs) in mitigating skin aging through multiple mechanisms have been widely reported. β-Nicotinamide mononucleotide (NMN) is an iconic anti-aging drug that increases NAD levels in the body to slow down, ameliorate, and prevent various phenotypes associated with aging, but its high water solubility, low permeability, and instability limit its clinical application. Based on this, we applied electroporation to construct NMN-loaded hucMSC-sEVs (NMN-sEVs) to improve their stability and efficacy and to enhance their potential for translational application in medical aesthetics and anti-aging.

METHODS

D-galactose was applied to construct a mouse skin aging model, based on which comparative analyses of topical and nano-microneedle administration were performed to determine the optimal delivery method of sEVs in vivo experiments. After constructing NMN-sEVs by electroporation, high-performance liquid chromatography was applied to detect the loading efficiency, and the effects of NMN-sEVs on delaying skin aging were assessed by histological analysis. In addition, the defense effects of NMN-sEVs against cellular senescence were verified by reactive oxygen species assay, β-galactosidase staining, qRT-PCR, Western blot, and cellular immunofluorescence. Finally, the roles of NMN-sEVs in remodeling mitochondrial function and delaying cellular senescence through mitochondrial autophagy were assessed by mitochondrial mass, function, and autophagy level assays.

RESULTS

Our data suggested that NMN-sEVs could improve skin aging in mice, delay cellular senescence, and restore cellular mitochondrial dysfunction. Notably, NMN-sEVs treatment increased intracellular NAD levels and SIRT3 expression, as well as rescued the inhibition of senescence-induced mitochondrial autophagy, suggesting a role for NMN-sEVs in the remodeling of mitochondrial function through mitochondrial autophagy. Additionally, the use of the SIRT3 inhibitor 3-TYP suppressed the positive effects of NMN-sEVs on cellular senescence, mitochondrial function, and mitochondrial autophagy while restoring senescence-associated characteristics.

CONCLUSION

Overall, our findings revealed a mechanism by which NMN-sEVs attenuated mitochondrial dysfunction and rescued cellular senescence by promoting NAD/SIRT3 pathway-mediated mitophagy and might provide a promising strategy for anti-aging pharmaceuticals.

摘要

背景

最近，人脐带间充质基质细胞（hucMSC）衍生的小细胞外囊泡（sEVs）通过多种机制减轻皮肤衰老的有益作用已被广泛报道。β-烟酰胺单核苷酸（NMN）是一种标志性的抗衰老药物，可提高体内NAD水平，以减缓、改善和预防与衰老相关的各种表型，但其高水溶性、低渗透性和不稳定性限制了其临床应用。基于此，我们应用电穿孔技术构建了负载NMN的hucMSC-sEVs（NMN-sEVs），以提高其稳定性和功效，并增强其在医学美容和抗衰老方面的转化应用潜力。

方法

应用D-半乳糖构建小鼠皮肤衰老模型，在此基础上进行局部给药和纳米微针给药的比较分析，以确定sEVs在体内实验中的最佳给药方式。通过电穿孔构建NMN-sEVs后，应用高效液相色谱法检测负载效率，并通过组织学分析评估NMN-sEVs对延缓皮肤衰老的作用。此外，通过活性氧测定、β-半乳糖苷酶染色、qRT-PCR、蛋白质免疫印迹和细胞免疫荧光验证了NMN-sEVs对细胞衰老的防御作用。最后，通过线粒体质量、功能和自噬水平测定评估了NMN-sEVs在通过线粒体自噬重塑线粒体功能和延缓细胞衰老中的作用。

结果

我们的数据表明，NMN-sEVs可以改善小鼠皮肤衰老，延缓细胞衰老，并恢复细胞线粒体功能障碍。值得注意的是，NMN-sEVs处理增加了细胞内NAD水平和SIRT3表达，并挽救了衰老诱导的线粒体自噬抑制，表明NMN-sEVs在通过线粒体自噬重塑线粒体功能中发挥作用。此外，使用SIRT3抑制剂3-TYP可抑制NMN-sEVs对细胞衰老、线粒体功能和线粒体自噬的积极作用，同时恢复衰老相关特征。