中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (7): 1768-1781.doi: 10.12307/2026.118

• 干细胞外泌体 Stem cell exosomes • 上一篇    下一篇

瓜蒌类外泌体囊泡防治动脉粥样硬化的作用及机制

陈钰璘1，2，何莹莹1，2，胡  凯1，2，陈枝凡1，2，聂  莎1，2，蒙衍慧1，2，李闰珍1，2，张小朵1，2，李宇稀2，唐耀平1，2，3   

  1. 1广西中医药大学，广西壮族自治区南宁市   530200；2广西中医药大学附属瑞康医院，广西壮族自治区南宁市   530200；3广西中医药大学国际教育学院，广西壮族自治区南宁市   530001
  • 收稿日期:2025-01-11 修回日期:2025-06-20 接受日期:2025-07-03 出版日期:2026-03-08 发布日期:2025-08-19
  • 通讯作者: 唐耀平，博士，教授，主任医师，广西中医药大学，广西壮族自治区南宁市 530200；广西中医药大学附属瑞康医院，广西壮族自治区南宁市 530200； 广西中医药大学国际教育学院，广西壮族自治区南宁市 530001
  • 作者简介:陈钰璘，男，1993年生，广西壮族自治区钦州市人，汉族，广西中医药大学在读硕士，主要从事心血管疾病的中西医结合防治研究。
  • 基金资助:
    国家自然科学基金委员会地区科学基金项目(82160856)，项目负责人：唐耀平；广西中医药大学硕士研究生创新项目(YCSW2024418)，项目负责人：陈钰璘

Effect and mechanism of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. in preventing and treating atherosclerosis

Chen Yulin1, 2, He Yingying1, 2, Hu Kai1, 2, Chen Zhifan1, 2, Nie Sha1, 2, Meng Yanhui1, 2, Li Runzhen1, 2, Zhang Xiaoduo1, 2, Li Yuxi2, Tang Yaoping1, 2, 3   

  1. 1Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 2Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 3Faculty of International Education, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China 
  • Received:2025-01-11 Revised:2025-06-20 Accepted:2025-07-03 Online:2026-03-08 Published:2025-08-19
  • Contact: Tang Yaoping, MD, Professor, Chief physician, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; Faculty of International Education, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • About author:Chen Yulin, Master candidate, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China (Regional Science Fund Project), No. 82160856 (to TYP); Guangxi University of Chinese Medicine Master Graduate Student Innovation Project, No. YCSW2024418 (to CYL)

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摘要：

文题释义：

NOD样受体家族Pyrin域蛋白3(NOD-like receptors family Pyrin domain containing 3，NLRP3)炎症小体：NLRP3蛋白、含有CARD结构域的凋亡相关斑点样蛋白以及Caspase-1共同组装形成一个多聚蛋白复合体，称为NLRP3炎性小体。NLRP3炎性小体能够激活Caspase-1，触发促炎性细胞因子的释放。
类外泌体囊泡：是一种由异质细胞群释放的膜性囊泡，包括来源于内体系统的外泌体和来源于质膜的微泡。它们通过将生物活性物质(如DNA、RNA、miRNA、蛋白质及其他代谢物)传递至靶细胞，从而触发细胞的生物学反应，是细胞间通信的重要递质。

摘要
背景：瓜蒌具有抗氧化功能和抗炎作用，常用于治疗心血管疾病。然而，瓜蒌的疗效往往受到成分复杂和生物利用度低的限制。
目的：探讨瓜蒌类外泌体囊泡防治动脉粥样硬化的生物学机制。
方法：①采用密度梯度离心法从瓜蒌中提取细胞外囊泡，通过透射电镜、纳米粒径检测仪与纳米颗粒追踪分析仪鉴定，共聚焦扫描显微镜观察THP-1来源巨噬细胞摄入瓜蒌类外泌体囊泡情况。②THP-1来源巨噬细胞随机分为空白组、模型组及瓜蒌类外泌体囊泡低、中、高剂量组，其中空白组正常培养，模型组加入50 mg/L氧化低密度脂蛋白，瓜蒌类外泌体囊泡低、中、高剂量组加入50 mg/L氧化低密度脂蛋白和5，10，20 mg/L瓜蒌类外泌体囊泡，干预24 h，采用Western blot检测细胞NLRP3和Cleaved-Caspase-1表达水平。③33只Apoe-/-小鼠随机分为空白组、模型组和瓜蒌类外泌体囊泡组，其中空白组小鼠喂养普通饲料，其余2组小鼠喂养高脂饲料16周后，瓜蒌类外泌体囊泡组小鼠腹腔注射瓜蒌类外泌体囊泡，空白组与模型组小鼠腹腔注射生理盐水，隔日给药1次，连续4周，采用油红O染色检测主动脉斑块面积，苏木精-伊红染色观察主动脉组织病理变化，ELISA检测血清白细胞介素1β、白细胞介素18水平，试剂盒检测血脂水平，免疫组化检测主动脉NLRP3和Cleaved-Caspase-1表达水平。④高效液相色谱-质谱联用技术与网络药理学预测瓜蒌类外泌体囊泡治疗冠心病的潜在靶点。

结果与结论：①瓜蒌类外泌体囊泡呈茶托状，粒径在50-150 nm之间，能被THP-1来源巨噬细胞有效摄入；②细胞实验显示，与模型组相比，瓜蒌类外泌体囊泡低、中、高剂量组NLRP3和Cleaved-Caspase-1表达水平呈浓度依赖型降低；③小鼠实验显示，与模型组相比，瓜蒌类外泌体囊泡组小鼠主动脉斑块面积减少，炎症细胞浸润较少，三酰甘油、总胆固醇、低密度脂蛋白胆固醇水平下降，高密度脂蛋白胆固醇水平升高，NLRP3及Cleaved-Caspase-1蛋白表达降低；血清白细胞介素1β、白细胞介素18水平降低；④筛选并富集到的关键通路有HIF-1信号通路、PI3K-Akt信号通路、代谢通路、脂质和动脉粥样硬化通路等。结果表明，瓜蒌类外泌体囊泡可有效缓解小鼠动脉粥样硬化，作用机制可能与抑制NLRP3炎症小体介导的炎症反应有关。

https://orcid.org/0000-0003-4840-1148 (唐耀平) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


关键词: 瓜蒌, 类外泌体囊泡, NLRP3炎症小体, 动脉粥样硬化, 炎症反应, 高效液相色谱-质谱, 网络药理学, 冠心病

Abstract: BACKGROUND: Trichosanthes kirilowii Maxim. possesses antioxidant and anti-inflammatory properties and is commonly used in the treatment of cardiovascular diseases. However, the therapeutic efficacy of Trichosanthes kirilowii Maxim. is often limited by its complex composition and low bioavailability.
OBJECTIVE: To investigate the biological mechanisms underlying the effects of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. in the prevention and treatment of atherosclerosis.
METHODS: (1) Extracellular vesicles were extracted from Trichosanthes kirilowii Maxim. using density gradient centrifugation. These vesicles were identified by transmission electron microscopy, nanoparticle size analyzer, and nanoparticle tracking analysis. Confocal laser scanning microscopy was used to observe the uptake of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. by THP-1 cells. (2) THP-1 derived macrophages were randomly divided into the blank group, model group, and low-, medium-, and high-dose exosome-like vesicles derived from Trichosanthes kirilowii Maxim. groups. The blank group was cultured under normal conditions; the model group was treated with 50 mg/L oxidized low-density lipoprotein, and the exosome-like vesicles derived from Trichosanthes kirilowii Maxim. groups were treated with 50 mg/L oxidized low-density lipoprotein and 5, 10, or 20 mg/L of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. After 24 hours of intervention, the expression levels of NLRP3 and Cleaved-Caspase-1 in the cells were assessed by western blot assay. (3) Thirty-three Apoe-/- mice were randomly divided into the blank group, model group, and exosome-like vesicles derived from Trichosanthes kirilowii Maxim. group. The blank group was fed a standard diet, while the other two groups were fed a high-fat diet for 16 weeks. Afterward,  the exosome-like vesicles derived from Trichosanthes kirilowii Maxim. group received intraperitoneal injections of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. The blank and model groups received intraperitoneal injections of physiological saline, every other day for 4 weeks. Oil Red O staining was used to assess aortic plaque area. Hematoxylin-eosin staining was performed to observe aortic tissue pathological changes. Enzyme-linked immunosorbent assay was used to measure serum interleukin-1β and interleukin-18 levels. A reagent kit was used to assess blood lipid levels. Immunohistochemistry was employed to detect the expression of NLRP3 and Cleaved-Caspase-1 in the aorta. (4) High-performance liquid chromatography coupled with mass spectrometry and network pharmacology were used to predict the potential therapeutic targets of exosome-like vesicles derived from Trichosanthes kirilowii Maxim. in the treatment of coronary heart disease.
RESULTS AND CONCLUSION: (1) The exosome-like vesicles derived from Trichosanthes kirilowii Maxim. exhibited a tea-tray-like shape, with a size range of 50–150 nm, and were effectively internalized by THP-1 derived macrophages. (2) Cell experiments showed that compared to the model group, the expression levels of NLRP3 and Cleaved-Caspase-1 in the low-, medium-, and high-dose exosomes derived from Trichosanthes kirilowii Maxim. groups were reduced in a dose-dependent manner. (3) Mouse experiments revealed that compared to the model group, the exosome-like vesicles derived from Trichosanthes kirilowii Maxim. group showed reduced aortic plaque area, less inflammatory cell infiltration, decreased triglycerides, total cholesterol, and low-density lipoprotein cholesterol, increased high-density lipoprotein cholesterol, and reduced expression of NLRP3 and Cleaved-Caspase-1 proteins. Additionally, serum interleukin-1β and interleukin-18 levels were decreased. (4) Key pathways identified and enriched included the HIF-1 signaling pathway, PI3K-Akt signaling pathway, metabolic pathways, lipid metabolism, and atherosclerosis pathways. These results suggest that exosome-like vesicles derived from Trichosanthes kirilowii Maxim. can effectively alleviate atherosclerosis in mice, and their mechanism of action may be related to the inhibition of NLRP3 inflammasome-mediated inflammatory responses.

Key words: Trichosanthes kirilowii Maxim., exosome-like vesicle, NLRP3 inflammasome, atherosclerosis, inflammatory response, high performance liquid chromatography-mass, network pharmacology, coronary heart disease 

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