中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (13): 3250-3257.doi: 10.12307/2026.088

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

人羊膜间充质干细胞外泌体修复放射性损伤SD大鼠颌下腺上皮细胞的功能

张立刚1，刘  涛2，易  杰1，张霓霓1，姚  礼1，黄桂林3，胡小华1，代  敏1   

  1. 1遵义医科大学口腔医学院·附属口腔医院·贵州省普通高等学校口腔疾病研究特色重点实验室，贵州省遵义市   566300；2内江市第一人民医院，四川省内江市   641000；3遵义医科大学珠海校区，广东省珠海市   519041
  • 收稿日期:2025-03-27 修回日期:2025-06-13 接受日期:2025-07-14 出版日期:2026-05-08 发布日期:2025-12-24
  • 通讯作者: 黄桂林，博士，主任医师，教授，遵义医科大学珠海校区，广东省珠海市 519041
  • 作者简介:张立刚，男，1987年生，硕士，副主任医师，副教授，主要从事唾液腺组织工程修复及放射性损伤唾液腺的干细胞治疗研究。
  • 基金资助:
    贵州省教育厅青年科技人才成长项目(黔教合KY字[2022]291号)，项目负责人：张立刚；贵州省卫生健康委科学技术基金项目(gzwjkj2020-1-165)，项目负责人：张立刚；贵州省临床重点建设项目(黔卫计办涵[2017]24号)，项目负责人：黄桂林；遵义医科大学“未来临床名医”(20211017)，项目负责人：张霓霓；遵义市科技计划项目(遵市科合HZ字(2023)87号)，项目负责人：张立刚

Function of human amniotic mesenchymal stem cell exosomes in repairing submandibular gland epithelial cells after radiation injury in SD rats

Zhang Ligang1, Liu Tao2, Yi Jie1, Zhang Nini1, Yao Li1, Huang Guilin3, Hu Xiaohua1, Dai Min1   

  1. 1School of Stomatology · Affiliated Stomatological Hospital · Key Laboratory of Oral Disease Research in Guizhou Province, Zunyi Medical University, Zunyi 566300, Guizhou Province, China; 2First People’s Hospital of Neijiang, Neijiang 641000, Sichuan Province, China; 3Zhuhai Campus of Zunyi Medical University, Zhuhai 519041, Guangdong Province, China
  • Received:2025-03-27 Revised:2025-06-13 Accepted:2025-07-14 Online:2026-05-08 Published:2025-12-24
  • Contact: Huang Guilin, MD, Chief physician, Professor, Zhuhai Campus of Zunyi Medical University, Zhuhai 519041, Guangdong Province, China
  • About author:Zhang Ligang, MS, Associate chief physician, Associate professor, School of Stomatology · Affiliated Stomatological Hospital · Key Laboratory of Oral Disease Research in Guizhou Province, Zunyi Medical University, Zunyi 566300, Guizhou Province, China
  • Supported by:
    Youth Science and Technology Talent Growth Project of Guizhou Provincial Department of Education, No. KY[2022]291 (to ZLG); Science and Technology Fund Project of Guizhou Provincial Health Commission, No. gzwjkj2020-1-165 (to ZLG); Guizhou Provincial Key Clinical Construction Project, No. [2017]24 (to HGL); “Future Clinical Famous Doctor” of Zunyi Medical University, No. 20211017 (to ZNN); Zunyi Science and Technology Plan Project, No. HZ(2023)87 (to ZLG) 

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摘要：

文题释义：

间充质干细胞：是一种具有自我更新能力的成体干细胞，广泛存在于骨髓和其他组织中。间充质干细胞来源外泌体具有与间充质干细胞相似的治疗作用，在组织修复和再生方面具有显著效果。
放射性唾液腺功能损伤：在接受辐射后唾液腺功能出现急性损伤并表现出相应的症状，主要表现为腺体肿胀、唾液量分泌降低、唾液分泌异常电解质及蛋白质等。

摘要
背景：低氧预处理能促进间充质干细胞旁分泌细胞外囊泡、生长因子、抗炎因子及免疫抑制相关因子等，低氧预处理的人羊膜间充质干细胞来源外泌体有望在组织损伤修复中发挥更好的治疗作用。
目的：观察低氧预处理的人羊膜间充质干细胞外泌体对放射性颌下腺上皮细胞损伤的修复作用。
方法：将第3代人羊膜间充质干细胞分为低氧及常氧两组，分别给予低氧(体积分数1%O2)及常氧(体积分数20%O2)预处理 48 h，然后通过改良超高速离心法从人羊膜间充质干细胞培养上清液中提取常氧和低氧人羊膜间充质干细胞外泌体。将SD新生鼠颌下腺上皮细胞分为空白对照组、放射对照组、常氧加药组、低氧加药组，给予颌下腺上皮细胞5 Gy放射处理，构建放射性损伤颌下腺上皮细胞模型，常氧加药组、低氧加药组在此基础上加入常氧或低氧人羊膜间充质干细胞外泌体与颌下腺上皮细胞共培养3 d。CCK-8法检测各组大鼠颌下腺上皮细胞的增殖活性，ELISA 测定各组大鼠颌下腺上皮细胞上清液中α-淀粉酶含量；RT-qPCR 检测各组大鼠颌下腺上皮细胞中水通道蛋白5 mRNA表达。
结果与结论：①放射对照组的细胞增殖活性较空白对照组明显下降(P < 0.05)；常氧加药组和低氧加药组第2，3天的细胞增殖活性明显高于放射对照组(P < 0.05)；低氧加药组第1天的细胞增殖活性高于常氧加药组，第2，3天有升高趋势，但与常氧加药组相比无统计学差异(P > 0.05)；②放射对照组α-淀粉酶含量较空白对照组显著降低(P < 0.05)；常氧加药组、低氧加药组α-淀粉酶含量较放射对照组显著升高(P < 0.05)；与常氧加药组相比，低氧加药组α-淀粉酶含量有升高趋势，但无统计学差异(P > 0.05)；③放射对照组水通道蛋白5 mRNA表达较空白对照组显著降低(P < 0.05)；常氧加药组、低氧加药组水通道蛋白5 mRNA表达较放射对照组显著升高(P < 0.05)；低氧加药组水通道蛋白5 mRNA表达较常氧加药组稍升高，但无统计学差异(P > 0.05)。结果表明：常氧和低氧预处理的人羊膜间充质干细胞外泌体对放射性颌下腺上皮细胞损伤均有修复作用，但低氧人羊膜间充质干细胞外泌体无明显优势。

关键词: 人羊膜间充质干细胞, 放射性损伤, 颌下腺上皮细胞, 外泌体, α-淀粉酶, 水通道蛋白5, 低氧预处理

Abstract: BACKGROUND: Hypoxia preconditioning can increase extracellular vesicles, growth factors, anti-inflammatory and immunosuppressive factors in the parasecretory secretions of mesenchymal stem cells. Exosomes derived from human amniotic mesenchymal stem cells preconditioned with hypoxia are expected to play a better therapeutic role in tissue damage repair.
OBJECTIVE: To observe the repair effect of exosomes from human amniotic mesenchymal stem cells preconditioned with hypoxia on radiation-induced submandibular gland epithelial cell damage. 
METHODS: The passage 3 human amniotic mesenchymal stem cells were divided into two groups: hypoxia and normoxia. They were pretreated with hypoxia (1% O2 by volume) and normoxia (20% O2 by volume) for 48 hours, respectively. Then, normoxic and hypoxic human amniotic mesenchymal stem cell exosomes were extracted from the culture supernatant of human amniotic mesenchymal stem cells by modified ultrahigh-speed centrifugation. The submandibular gland epithelial cells of SD newborn rats were divided into blank control group, radiation control group, normoxic drug group, and hypoxic drug group. The submandibular gland epithelial cells were irradiated with 5 Gy to establish the radiation-damaged submandibular gland epithelial cell model. On this basis, normoxic and hypoxic human amniotic mesenchymal stem cell exosomes were added to the normoxic drug group and the hypoxic drug group and co-cultured with the submandibular gland epithelial cells for 3 days. The proliferation activity of the submandibular gland epithelial cells of the rats in each group was detected by CCK-8 assay. The alpha amylase content in the supernatant of the submandibular gland epithelial cells of the rats in each group was determined by ELISA. RT-qPCR was used to detect the expression of aquaporin 5 mRNA in the submandibular gland epithelial cells of the rats in each group.
RESULTS AND CONCLUSION: (1) The cell proliferation activity of the radiation control group was significantly decreased compared to the blank control group (P < 0.05). The cell proliferation activity of the normoxic and hypoxic drug groups significantly increased on days 2 and 3 compared to the radiation control group (P < 0.05). The cell proliferation activity of the hypoxic drug group was higher than that of the hypoxic drug group on the first day, and showed an increasing trend on the second and third days, but there was no statistical difference compared with the normoxic drug group (P > 0.05). (2) The content of alpha amylase in the radiation control group was significantly reduced compared with the blank control group (P < 0.05). The levels of alpha amylase in the normoxic and hypoxic drug groups were significantly higher than those in the radiation control group (P < 0.05). Compared with the normoxic drug group, the hypoxic drug group showed an increasing trend in alpha amylase content, but there was no statistical difference (P > 0.05). (3) The mRNA expression of aquaporin 5 in the radiation control group was significantly reduced compared to the blank control group (P < 0.05). The mRNA expression of aquaporin 5 in the normoxic and hypoxic drug groups was significantly increased compared with the radiation control group (P < 0.05). The mRNA expression of aquaporin 5 in the hypoxic drug group was slightly higher than that in the normoxic drug group, but there was no statistical difference (P > 0.05). These results indicate that extracellular vesicles from human amniotic mesenchymal stem cells pre-treated with normoxic and hypoxic conditions have a repairing effect on submandibular gland epithelial cells damaged by radiation, but there is no significant advantage in hypoxic human amniotic mesenchymal stem cell exosomes. 

Key words: human amniotic mesenchymal stem cell, radiation damage, submandibular gland epithelial cell, exosome, alpha amylase, aquaporin 5, hypoxic preconditioning

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