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已发表论文
姜来源的外泌体样纳米颗粒：提取方法对代谢物及体外抗肺癌活性的影响
Authors Ming T, Yang Y, Zhu J, Lin J, Yang W, Yu G, Wang P, Zhang E, Chen Q, Liu J
Received 21 May 2025
Accepted for publication 22 October 2025
Published 6 November 2025 Volume 2025:20 Pages 13399—13420
DOI https://doi.org/10.2147/IJN.S541948
Checked for plagiarism Yes
Review by Single anonymous peer review
Peer reviewer comments 5
Editor who approved publication: Professor Farooq A. Shiekh
Tingwen Ming,1,* Yang Yang,2,* Jun Zhu,1 Juncheng Lin,1 Wang Yang,1 Guanbin Yu,1 Pengju Wang,1 Enxin Zhang,2 Qinhua Chen,2 Jingjian Liu1,3
1Sinopharm Dongfeng General Hospital, Hubei University of Medicine, Shiyan, 442008, People’s Republic of China; 2Shenzhen Bao’an Authentic TCM Therapy Hospital, Shenzhen, 518101, People’s Republic of China; 3Hubei Key Laboratory of Wudang Local Chinese Medicine Research, Hubei University of Medicine, Shiyan, 442000, People’s Republic of China
*These authors contributed equally to this work
Correspondence: Qinhua Chen, Email cqh77@163.com Jingjian Liu, Email liujj95@163.com
Introduction: In recent years, plant-derived exosome-like nanoparticles (PELNs) have attracted extensive attention. Among them, Ginger-derived exosome-like nanoparticles (GELNs) represent the most extensively studied category, demonstrating a wide spectrum of pharmacological activities. However, their specific efficacy against lung cancer remains largely unexplored and warrants further investigation. The appropriate isolation of GELNs is fundamental to all related research, yet a systematic comparison of different extraction methods is currently lacking. This study aimed to evaluate the differences among GELNs extracted by various methods and to investigate their anti-lung cancer pharmacological activities.
Methods: The study employed four common isolation methods—ultracentrifugation (UC), sucrose gradient UC (sgUC), membrane filtration, and polyethylene glycol-based precipitation (PEG-based precipitation) - to isolate GELNs. The GELNs were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and zeta potential measurements. Stability was evaluated under various conditions, including saline, serum, and different storage temperatures. The compositional profiles of GELNs extracted by four methods were explored using non-targeted metabolomics. A549 cells and PC-9 cells were used to assess the cellular uptake and anti-lung cancer efficacy of the four GELNs types. Network pharmacology, molecular docking, and molecular dynamics simulations were integrated to elucidate the potential mechanisms underlying their anti-lung cancer effects.
Results: The four methods successfully isolated GELNs with distinct profiles: UC achieved the highest protein yield (1.630 ± 0.022 g/kg), membrane filtration yielded the highest particle concentration (46.9 ± 6.71× 108 particles/mL) but the lowest protein yield (0.059 ± 0.002 g/kg). Stability studies indicated that the highest stability of GELNs was observed for those isolated by UC and sgUC in both 0.9% and 10% NaCl. Furthermore, GELNs prepared by UC and membrane filtration showed excellent stability in serum. It was also demonstrated that − 80°C provided the optimal storage condition for GELNs. Non-targeted metabolomics revealed the presence of 649 shared metabolites among the GELNs extracted by the four methods, along with method-specific unique metabolites. GELNs extracted by all four methods were internalized by both A549 and PC-9 cells. Among them, UC-isolated GELNs demonstrated the most potent anti-proliferative activity against the lung cancer cells. Through network pharmacology, 21 key targets of UC-isolated GELNs against lung cancer were identified. Molecular docking and molecular dynamics simulations further verified that 10-Gingerol, Hexahydrocurcumin, and [6]-Dehydrogingerdione from GELNs could stably bind to key targets, including Glycogen Synthase Kinase-3β (GSK3B), Progesterone Receptor (PGR), and SRC Proto-Oncogene, Non-Receptor Tyrosine Kinase (SRC).
Conclusion: This study demonstrates that although all four methods can isolate GELNs, UC is recommended for fundamental research due to its high protein yield, excellent stability, and potent in vitro anti-lung cancer activity. Furthermore, the anti-lung cancer activity of GELNs may be attributed to the regulation of GSK3B, PGR, and SRC by 10-Gingerol, Hexahydrocurcumin, and [6]-Dehydrogingerdione.
Keywords: ginger-derived exosome-like nanoparticles, extraction method, stability, metabolomics, lung cancer, network pharmacology
1Sinopharm Dongfeng General Hospital, Hubei University of Medicine, Shiyan, 442008, People’s Republic of China; 2Shenzhen Bao’an Authentic TCM Therapy Hospital, Shenzhen, 518101, People’s Republic of China; 3Hubei Key Laboratory of Wudang Local Chinese Medicine Research, Hubei University of Medicine, Shiyan, 442000, People’s Republic of China
*These authors contributed equally to this work
Correspondence: Qinhua Chen, Email cqh77@163.com Jingjian Liu, Email liujj95@163.com
Introduction: In recent years, plant-derived exosome-like nanoparticles (PELNs) have attracted extensive attention. Among them, Ginger-derived exosome-like nanoparticles (GELNs) represent the most extensively studied category, demonstrating a wide spectrum of pharmacological activities. However, their specific efficacy against lung cancer remains largely unexplored and warrants further investigation. The appropriate isolation of GELNs is fundamental to all related research, yet a systematic comparison of different extraction methods is currently lacking. This study aimed to evaluate the differences among GELNs extracted by various methods and to investigate their anti-lung cancer pharmacological activities.
Methods: The study employed four common isolation methods—ultracentrifugation (UC), sucrose gradient UC (sgUC), membrane filtration, and polyethylene glycol-based precipitation (PEG-based precipitation) - to isolate GELNs. The GELNs were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and zeta potential measurements. Stability was evaluated under various conditions, including saline, serum, and different storage temperatures. The compositional profiles of GELNs extracted by four methods were explored using non-targeted metabolomics. A549 cells and PC-9 cells were used to assess the cellular uptake and anti-lung cancer efficacy of the four GELNs types. Network pharmacology, molecular docking, and molecular dynamics simulations were integrated to elucidate the potential mechanisms underlying their anti-lung cancer effects.
Results: The four methods successfully isolated GELNs with distinct profiles: UC achieved the highest protein yield (1.630 ± 0.022 g/kg), membrane filtration yielded the highest particle concentration (46.9 ± 6.71× 108 particles/mL) but the lowest protein yield (0.059 ± 0.002 g/kg). Stability studies indicated that the highest stability of GELNs was observed for those isolated by UC and sgUC in both 0.9% and 10% NaCl. Furthermore, GELNs prepared by UC and membrane filtration showed excellent stability in serum. It was also demonstrated that − 80°C provided the optimal storage condition for GELNs. Non-targeted metabolomics revealed the presence of 649 shared metabolites among the GELNs extracted by the four methods, along with method-specific unique metabolites. GELNs extracted by all four methods were internalized by both A549 and PC-9 cells. Among them, UC-isolated GELNs demonstrated the most potent anti-proliferative activity against the lung cancer cells. Through network pharmacology, 21 key targets of UC-isolated GELNs against lung cancer were identified. Molecular docking and molecular dynamics simulations further verified that 10-Gingerol, Hexahydrocurcumin, and [6]-Dehydrogingerdione from GELNs could stably bind to key targets, including Glycogen Synthase Kinase-3β (GSK3B), Progesterone Receptor (PGR), and SRC Proto-Oncogene, Non-Receptor Tyrosine Kinase (SRC).
Conclusion: This study demonstrates that although all four methods can isolate GELNs, UC is recommended for fundamental research due to its high protein yield, excellent stability, and potent in vitro anti-lung cancer activity. Furthermore, the anti-lung cancer activity of GELNs may be attributed to the regulation of GSK3B, PGR, and SRC by 10-Gingerol, Hexahydrocurcumin, and [6]-Dehydrogingerdione.
Keywords: ginger-derived exosome-like nanoparticles, extraction method, stability, metabolomics, lung cancer, network pharmacology