植物源外泌体样纳米囊泡抗衰老调控及工程化研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2026-0253

摘要/Abstract

摘要：

植物源外泌体样纳米囊泡（plant-derived exosome-like nanovesicles, PENs）是从水果、蔬菜及药用植物中分离获得的一类天然纳米级囊泡，直径约为30‒300 nm，具备典型的脂质双层膜结构。作为来源广泛、生物相容性良好且免疫原性较低的天然递送载体，PENs在抗衰老研究领域受到越来越多的关注。本文系统综述了PENs的生物学基础、抗衰老调控机制及工程化修饰策略的研究进展。PENs自身携带脂质、蛋白质、核酸及次生代谢产物等物质，其抗衰老活性主要归因于对氧化应激、炎症反应、细胞外基质代谢、细胞衰老与自噬以及黑色素生成等多条生物学通路的调节。面对天然PENs在靶向能力、载荷效率及体内循环时间等方面存在的不足，研究人员开发了被动与主动装载、聚乙二醇化修饰、配体偶联及膜成分调控等多种工程化策略，显著提升其递送效能与治疗精准度。此外，本文指出PENs的临床转化仍面临多重挑战，包括现有临床试验样本量偏小且缺乏严谨的设计规范，部分植物来源可能伴有潜在毒性风险，量效关系研究尚不充分，监管框架与标准化体系仍属空白，制备工艺和长期保存技术也有待进一步完善等。未来建立标准化、自动化的生产系统与统一的质量控制体系，开展设计严谨的剂量探索性临床试验，将是推动PENs在抗衰老领域深入应用和临床落地的关键路径。本文为植物源外泌体样纳米囊泡在抗衰老及相关疾病治疗中的深入研究提供了系统性的理论参考。

关键词: 植物源外泌体样纳米囊泡, 抗衰老, 调控机制, 工程化改造, 靶向递送, 氧化应激, 纳米载体, 光老化

Abstract:

Plant-derived exosome-like nanovesicles (PENs) are a class of natural nanoscale vesicles isolated from fruits, vegetables, and medicinal plants, with diameters of 30-300 nm and a characteristic lipid bilayer membrane structure. As naturally occurring delivery vehicles that are widely available, biocompatible, and weakly immunogenic, PENs have garnered growing interest in anti-aging research. This review systematically summarizes recent progress in the biological foundations, anti-aging regulatory mechanisms, and engineering modification strategies of PENs. PENs naturally carry lipids, proteins, nucleic acids, and secondary metabolites. Their anti-aging effects mainly arise from modulating multiple biological pathways, including oxidative stress, inflammation, extracellular matrix metabolism, cellular senescence and autophagy, and melanin production. To overcome the limitations of native PENs—such as poor targeting ability, low loading efficiency, and short circulation time—various engineering strategies have been developed, including passive and active loading, PEGylation, ligand conjugation, and membrane composition modulation. These approaches significantly improve delivery efficiency and therapeutic precision. Nevertheless, clinical translation of PENs still faces considerable challenges. Current clinical trials often have small sample sizes and lack rigorous design; some plant sources may pose potential toxicity risks; dose-response relationships remain insufficiently explored; regulatory frameworks and standardization systems are absent; and manufacturing processes and long-term storage techniques need further optimization. Establishing standardized, automated production systems and unified quality control protocols, along with well-designed dose-finding clinical trials, will be key to advancing the broader application and clinical adoption of PENs in anti-aging medicine. This review offers a comprehensive theoretical reference for future studies on PENs in anti-aging and related disease therapies.

Key words: plant-derived exosome-like nanovesicles, anti-aging, regulatory mechanisms, engineering modifications, targeted delivery, oxidative stress, nanocarriers, photoaging

雷崇彬, 粟元, 徐蒙, 朱龙佼, 商颖, 田洪涛, 许文涛. 植物源外泌体样纳米囊泡抗衰老调控及工程化研究进展[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0253.

LEI Chong-bin, SU Yuan, XU Meng, ZHU Long-jiao, SHANG Ying, TIAN Hong-tao, XU Wen-tao. Recent Advances in the Regulation and Engineering of Plant-derived Exosome-like Nanovesicles for Anti-aging[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0253.

图/表 5

图1 植物源外泌体样纳米囊泡（PENs）的生物学特征、抗衰老相关分子机制及多元化工程构建策略中央PENs图标代表核心纳米载体，可封装多种生物活性分子（黄酮类、miRNA、蛋白质、脂质）。内环圆展示了PENs的5大抗衰老机制，包括氧化应激调控、炎症反应抑制、细胞外基质代谢调节、衰老与自噬调控及黑色素生成调控。外环圆为PENs的四大工程化修饰策略，涵盖药物装载、靶向递送增强、稳定性与控释优化及免疫调节修饰。BPN：细菌‒植物杂交囊泡；PDE：磷酸二酯酶；CXCR2：CXC趋化因子受体2；LFA-1：淋巴细胞功能相关抗原1；siRNA：小干扰RNA；cAMP：环磷酸腺苷；PKA：蛋白激酶A；CREB：cAMP反应元件结合蛋白；MITF：小眼畸形相关转录因子；NF-κB：核因子κB；ERK1/2：细胞外信号调节激酶1/2；TNF-α：肿瘤坏死因子α；IL-6：白细胞介素6；IL-10：白细胞介素10；SA-β-Gal：衰老相关β-半乳糖苷酶；AMPK：AMP活化蛋白激酶；SIRT1：沉默信息调节因子1；PGC-1α：过氧化物酶体增殖物激活受体γ辅激活因子1α；CRY2：隐花色素2；cRGD：环状精氨酸-甘氨酸-天冬氨酸肽

Fig. 1 Biological characteristics, anti-aging-related molecular mechanisms, and diversified engineering strategies of plant-derived exosome-like nanovesicles (PENs)The central PENs icon represents the core nanocarriers, which can encapsulate various bioactive molecules (flavonoids, miRNAs, proteins, lipids). The inner circular diagram illustrates the five major anti-aging mechanisms of PENs, including oxidative stress regulation, inflammation response inhibition, extracellular matrix metabolism regulation, aging and autophagy regulation, and melanin production regulation. The outer circular diagram presents the four engineering modification strategies of PENs, covering drug loading, enhanced targeted delivery, stability and controlled release optimization, and immune regulation modification. BPN: Bacterial-plant hybrid nanovesicles. PDE: Phosphodiesterase. CXCR2: CXC chemokine receptor 2. LFA-1: Lymphocyte function-associated antigen 1. siRNA: Small interfering RNA. cAMP: Cyclic adenosine monophosphate. PKA: Protein kinase A. CREB: cAMP response element-binding protein. MITF: Microphthalmia-associated transcription factor. NF-κB: Nuclear factor kappa-light-chain-enhancer of activated B cells. ERK1/2: Extracellular signal-regulated kinase 1/2. TNF-α: Tumor necrosis factor-α. IL-6: Interleukin-6. IL-10: Interleukin-10. SA-β-Gal: Senescence-associated β-galactosidase. AMPK: AMP-activated protein kinase. SIRT1: Silent information regulator 1. PGC-1α: Peroxisome proliferator-activated receptor γ coactivator 1α. CRY2: Cryptochrome 2. cRGD: Cyclic arginine-glycine-aspartic acid peptide

图2 植物源外泌体样纳米囊泡介导抗氧化、抗炎、基质保护及抗黑色素生成的多靶点抗衰老分子调控机制图示总结了PENs通过5大途径调控衰老相关通路的分子机制：①氧化应激中，PENs激活AhR/Nrf2信号轴，促进Nrf2核转位并上调SOD2/GPX表达以清除ROS；②炎症中，PENs抑制NF-κB和ERK1/2通路，下调TNF-α/IL-6并上调IL-10；③ECM代谢中，PENs促进COL-1合成并抑制MMP-1以维持ECM稳态；④细胞衰老与自噬中，PENs（含miR-CM）激活AMPK/SIRT1/PGC-1α轴，促进自噬体形成并下调p21/SA-β-Gal；⑤黑色素生成中，PENS阻断α-MSH/MC1R/cAMP/PKA/CREB信号级联，抑制MITF核转位及TYR/TRP-1活性，减少黑素体中的黑色素合成。AhR：芳香烃受体；Nrf2：核因子E2相关因子2；SOD2：超氧化物歧化酶2；GPX：谷胱甘肽过氧化物酶；NF-κB：核因子κB；ERK1/2：细胞外信号调节激酶1/2；MAPK：丝裂原活化蛋白激酶；TNF-α：肿瘤坏死因子α；IL-6：白细胞介素6；IL-10：白细胞介素10；miR-CM1：微小RNA-CM1；AMPK：AMP活化蛋白激酶；SIRT1：沉默信息调节因子1；PGC-1α：过氧化物酶体增殖物激活受体γ辅激活因子1α；p21：周期蛋白依赖性激酶抑制剂p21；SA-β-Gal：衰老相关β-半乳糖苷酶；α-MSH：α-促黑素细胞激素；MC1R：黑皮质素1受体；CREB：cAMP反应元件结合蛋白；MITF：小眼畸形相关转录因子；TYR：酪氨酸酶；TRP-1：酪氨酸酶相关蛋白1；BPN：细菌-植物杂交囊泡；PDE：磷酸二酯酶

Fig. 2 Multi-target anti-aging molecular regulatory mechanisms of plant-derived exosome-like nanovesicles, involving anti-oxidation, anti-inflammation, ECM protection, and anti-melanogenesisThe figure summarizes the molecular mechanisms by which PENs regulates aging-related pathways through five major pathways: ① In oxidative stress, PENs activates the AhR/Nrf2 signaling axis, promoting Nrf2 nuclear translocation and upregulating SOD2/GPX expression to eliminate ROS. ② In inflammation, PENs suppresses the NF-κB and ERK1/2 pathways, downregulating TNF-α/IL-6 and upregulating IL-10. ③ In ECM metabolism, PENs promotes COL-1 synthesis and inhibits MMP-1 to maintain ECM homeostasis. ④ In cellular senescence and autophagy, PENs (containing miR-CM) activates the AMPK/SIRT1/PGC-1α axis, promoting autophagosome formation and downregulating p21/SA-β-Gal. ⑤ In melanogenesis, PENs blocks the α-MSH/MC1R/cAMP/PKA/CREB signaling cascade, inhibiting MITF nuclear translocation and TYR/TRP-1 activity, thereby reducing melanin synthesis in melanosomes. AhR: Aryl hydrocarbon receptor. Nrf2: Nuclear factor erythroid 2-related factor 2. SOD2: Superoxide dismutase 2. GPX: Glutathione peroxidase. NF-κB: Nuclear factor kappa-light-chain-enhancer of activated B cells. ERK1/2: Extracellular signal-regulated kinase 1/2. MAPK: Mitogen-activated protein kinase. TNF-α: Tumor necrosis factor-α. IL-6: Interleukin-6. IL-10: Interleukin-10. miR-CM1: MicroRNA-CM1. AMPK: AMP-activated protein kinase. SIRT1: Silent information regulator 1. PGC-1α: Peroxisome proliferator-activated receptor γ coactivator 1α. p21: Cyclin-dependent kinase inhibitor p21. SA-β-Gal: Senescence-associated β-galactosidase. α-MSH: α-Melanocyte-stimulating hormone. MC1R: Melanocortin 1 receptor. CREB: cAMP response element-binding protein. MITF: Microphthalmia-associated transcription factor. TYR: Tyrosinase. TRP-1: Tyrosinase-related protein 1. BPN: Bacteria-plant hybrid nanovesicles. PDE: Phosphodiesterase

表1 不同植物来源PENs的抗衰老功能比较

Table 1 Comparison of the anti-ageing functions of PENs from different plant sources

植物来源 Plant origin	代表性活性成分/搭载成分 Representative active ingredient / Loaded components	主要抗衰老机制 Main anti-aging mechanism	特殊优势 Special advantages	文献 References
柑橘Citrus reticulata Blanco.	类黄酮	激活AhR/Nrf2通路清除ROS；调节ERK1/2和NF-κB降低IFN-γ、TNF-α	直接清除自由基，抗炎	［34，41，45］
葡萄 Vitis vinifera L.	白藜芦醇	直接清除ROS，抑制脂质过氧化	富含白藜芦醇，抗氧化强	［42］
艾蒿Artemisia argyi H. Lév. & Vaniot	MIR2916	通过TLR4通路缓解氧化应激	miRNA介导的抗氧化调控	［44］
生姜 Zingiber officinale Roscoe	miRNA	下调IL-6、IL-8，抗炎；调节肠道菌群	口服稳定，肠道抗炎	［46］
茶 Camellia sinensis （L.） O. Ktze.	多酚类	降低TNF-α、IL-6、IL-12，上调IL-10	双向调节炎症因子	［47］
食用甜菜 Beta vulgaris L.	甜菜红素、酚类	促进COL-1表达，抑制MMP-1，降低SA-β-Gal	抗光老化，促胶原合成	［53］
桑黄Phellinus igniarius	miR-CM1	抑制Mical2，促进COL1A2，抑制MMP-1，降低ROS/MDA/SA-β-Gal	跨界miRNA调控ECM	［50］
薰衣草Lavandula angustifolia Mill.	多种活性成分	减轻炎症、增强ECM完整性、促进DNA修复、减轻氧化应激	多机制协同抗光老化	［51］
中国韭葱Allium porrum L.	—	激活AMPK/SIRT1/PGC-1α轴促进自噬；抑制Akt/FoxO3a/Atrogin-1/MuRF1	改善肌管萎缩，促线粒体生物发生	［55］
人参Panax ginseng C. A. Mey.	—	抑制AP-1通路，下调MMP-2/3、COX-2、IL-6、p21。抑制凋亡通路	抗凋亡，抗炎，抗衰老基因调控	［56］
山药豆 Dioscorea opposita	—	抑制黑色素合成，促进细胞迁移和胶原生成	美白+修复双重功效	［39］
葡萄柚 Citrus paradisi	—	抑制黑色素生成（B16-F10细胞、斑马鱼）	美白	［61］
苍术 Atractylodis rhizama	—	下调Mitf，降低酪氨酸酶活性	阻断黑色素合成通路	［62］

表1 不同植物来源PENs的抗衰老功能比较

Table 1 Comparison of the anti-ageing functions of PENs from different plant sources

植物来源 Plant origin	代表性活性成分/搭载成分 Representative active ingredient / Loaded components	主要抗衰老机制 Main anti-aging mechanism	特殊优势 Special advantages	文献 References
柑橘Citrus reticulata Blanco.	类黄酮	激活AhR/Nrf2通路清除ROS；调节ERK1/2和NF-κB降低IFN-γ、TNF-α	直接清除自由基，抗炎	［34，41，45］
葡萄 Vitis vinifera L.	白藜芦醇	直接清除ROS，抑制脂质过氧化	富含白藜芦醇，抗氧化强	［42］
艾蒿Artemisia argyi H. Lév. & Vaniot	MIR2916	通过TLR4通路缓解氧化应激	miRNA介导的抗氧化调控	［44］
生姜 Zingiber officinale Roscoe	miRNA	下调IL-6、IL-8，抗炎；调节肠道菌群	口服稳定，肠道抗炎	［46］
茶 Camellia sinensis （L.） O. Ktze.	多酚类	降低TNF-α、IL-6、IL-12，上调IL-10	双向调节炎症因子	［47］
食用甜菜 Beta vulgaris L.	甜菜红素、酚类	促进COL-1表达，抑制MMP-1，降低SA-β-Gal	抗光老化，促胶原合成	［53］
桑黄Phellinus igniarius	miR-CM1	抑制Mical2，促进COL1A2，抑制MMP-1，降低ROS/MDA/SA-β-Gal	跨界miRNA调控ECM	［50］
薰衣草Lavandula angustifolia Mill.	多种活性成分	减轻炎症、增强ECM完整性、促进DNA修复、减轻氧化应激	多机制协同抗光老化	［51］
中国韭葱Allium porrum L.	—	激活AMPK/SIRT1/PGC-1α轴促进自噬；抑制Akt/FoxO3a/Atrogin-1/MuRF1	改善肌管萎缩，促线粒体生物发生	［55］
人参Panax ginseng C. A. Mey.	—	抑制AP-1通路，下调MMP-2/3、COX-2、IL-6、p21。抑制凋亡通路	抗凋亡，抗炎，抗衰老基因调控	［56］
山药豆 Dioscorea opposita	—	抑制黑色素合成，促进细胞迁移和胶原生成	美白+修复双重功效	［39］
葡萄柚 Citrus paradisi	—	抑制黑色素生成（B16-F10细胞、斑马鱼）	美白	［61］
苍术 Atractylodis rhizama	—	下调Mitf，降低酪氨酸酶活性	阻断黑色素合成通路	［62］

图3 植物源外泌体样纳米囊泡（PENs）在药物负载、靶向递送、免疫调节及稳定性优化方面的工程化修饰策略图示总结了提升PENs的治疗性能的4大工程化策略：①药物装载：包括被动共孵育装载疏水性药物和主动超声/电穿孔装载siRNA/miRNA；②靶向修饰：通过EDC/NHS共价偶联cRGD、DSPE脂质锚定插入适配体R11-3、FA-PEG-Chol疏水插入叶酸等方式实现向肿瘤和脑组织的精准递送；③免疫工程修饰：包括FAP表达型PENs激活T细胞分化为CTLs，以及中性粒细胞膜包被的PENS表面展示CXCR2/LFA-1受体靶向炎症组织；④稳定性与控释增强：通过PEG化实现免疫逃逸和长循环，并结合近红外激光、pH敏感i-motif DNA和蓝光调控的CRY2/CIB系统实现智能触发释放。PENs：植物来源外泌体样纳米囊泡；ROS：活性氧；SOD2：超氧化物歧化酶2；GPX：谷胱甘肽过氧化物酶；AhR：芳香烃受体；Nrf2：核因子E2相关因子2；NF-κB：核因子κB；ERK1/2：细胞外信号调节激酶1/2；MAPK：丝裂原活化蛋白激酶；TNF-α：肿瘤坏死因子α；IL-6：白细胞介素6；IL-10：白细胞介素10；miR-CM1：微小RNA-CM1；COL-1：I型胶原蛋白；ECM：细胞外基质；MMP-1：基质金属蛋白酶1；AMPK：AMP活化蛋白激酶；SIRT1：沉默信息调节因子1；PGC-1α：过氧化物酶体增殖物激活受体γ辅激活因子1α；p21：周期蛋白依赖性激酶抑制剂p21；SA-β-Gal：衰老相关β-半乳糖苷酶；α-MSH：α-促黑素细胞激素；MC1R：黑皮质素1受体；CREB：cAMP反应元件结合蛋白；MITF：小眼畸形相关转录因子；TYR：酪氨酸酶；TRP-1：酪氨酸酶相关蛋白1

Fig. 3 Engineering modification strategies of plant-derived exosome-like nanovesicles (PENs) in drug loading, targeted delivery, immunomodulation and stability optimizationThis diagram summarizes four major engineering approaches to enhance PENs therapeutic performance: ① Cargo loading includes passive co-incubation for hydrophobic drugs and active sonication/electroporation for siRNA/miRNA encapsulation; ② Targeting modification achieves precise delivery to tumors and brain tissue via EDC/NHS covalent coupling of cRGD, DSPE lipid anchor insertion of aptamer R11-3, and FA-PEG-Chol hydrophobic insertion of folic acid; ③ Immune engineering involves FAP-expressing PENs activating T cells into CTLs and neutrophil membrane-coated PENs displaying CXCR2/LFA-1 receptors for inflamed tissue homing; ④ Stability and controlled release utilizes PEGylation for immune evasion and prolonged circulation, combined with NIR laser, pH-sensitive i-motif DNA, and blue light-controlled CRY2/CIB systems for smart triggered release. PENs: Plant-derived exosome-like nanovesicles. ROS: Reactive oxygen species. SOD2: Superoxide dismutase 2. GPX: Glutathione peroxidase. AhR: Aryl hydrocarbon receptor. Nrf2: Nuclear factor erythroid 2-related factor 2. NF-κB: Nuclear factor kappa-light-chain-enhancer of activated B cells. ERK1/2: Extracellular signal-regulated kinase 1/2. MAPK: Mitogen-activated protein kinase. TNF-α: Tumor necrosis factor-α. IL-6: Interleukin-6. IL-10: Interleukin-10. miR-CM1: MicroRNA-CM1. COL-1: Collagen type 1. ECM: Extracellular matrix. MMP-1: Matrix metalloproteinase 1. AMPK: AMP-activated protein kinase. SIRT1: Silent information regulator 1. PGC-1α: Peroxisome proliferator-activated receptor γ coactivator 1α. p21: Cyclin-dependent kinase inhibitor p21. SA-β-Gal: Senescence-associated β-galactosidase. α-MSH: α-Melanocyte-stimulating hormone. MC1R: Melanocortin 1 receptor. CREB: cAMP response element-binding protein. MITF: Microphthalmia-associated transcription factor. TYR: Tyrosinase. TRP-1: Tyrosinase-related protein 1

表2 不同工程化策略的优缺点

Table 2 Advantages and disadvantages of different engineering strategies

策略 Strategy	主要方法 Main methods	优势 Advantages	局限性 Limitations	参考文献 References
提高药物负载能力 Improve drug loading capacity	被动装载	操作简便；不破坏囊泡膜完整性	装载效率低	［92］
提高药物负载能力 Improve drug loading capacity	主动装载	装载效率高，适合核酸及亲水性大分子	可能影响囊泡稳定性；操作较复杂	［92］
改进靶向能力 Improve targeting ability	共价偶联（EDC/NHS）	连接稳定；靶向特异性高	可能影响表面天然蛋白功能	［79］
	脂质嵌入（DSPE+点击化学）	对膜结构干扰小；适配体亲和力高	适配体稳定性需优化
	疏水插入（FA-PEG-Chol）	操作简单；无需化学活化	插入稳定性受膜流动性影响
稳定性与缓释性能增强 Enhance stability and sustained-release performance	PEG化修饰	延长循环半衰期；降低免疫原性	过度PEG化可能降低靶向能力	［93］
	智能响应性递送	可控释放	需外部光源或特定微环境条件	［93］
免疫调节功能增强 Enhance immune regulation function	抗体偶联（双特异性抗体）	激活特异性免疫应答；实现免疫靶向	偶联效率需优化；潜在免疫原性	［94］
免疫调节功能增强 Enhance immune regulation function	杂交/融合策略（膜融合）	整合不同来源囊泡优势；增强归巢及免疫激活	融合效率及杂合囊泡稳定性控制复杂	［94］

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