口腔医学研究

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (12): 1145-1150.DOI: 10.13701/j.cnki.kqyxyj.2022.12.009

• 涎腺疾病研究 • 上一篇    下一篇

低氧预处理人羊膜间充质干细胞外泌体修复放射性损伤唾液腺的实验研究

崔田宁1, 张霓霓1, 龙远铸2, 黄桂林3*, 张立刚1, 唐建宏1, 骆勤亮1   

  1. 1.遵义医科大学附属口腔医院 贵州 遵义 563003;
    2.遵义医科大学 贵州 遵义 563000;
    3.遵义医科大学第五附属(珠海)医院 广东 珠海 519180
  • 收稿日期:2022-05-20 出版日期:2022-12-28 发布日期:2022-12-26
  • 通讯作者: *黄桂林,E-mail:chaojiehuanghgl@163.com
  • 作者简介:崔田宁(1995~ ),女,河南新乡人,硕士在读,主要从事放射性组织(涎腺)损伤修复研究及口腔颌面外科疾病的诊治。
  • 基金资助:
    国家自然科学基金(编号:81960204,81860198);贵州省卫健委基金项目(编号:gzwjkj2020-1-165)

Repair of Salivary Gland Radiation Injury by Exosomes from Human Amniotic Mesenchymal Stem Cells Pretreated with Hypoxia

CUI Tianning1, ZHANG Nini1, LONG Yuanzhu2, HUANG Guilin3*, ZHANG Ligang1, TANG Jianhong1, LUO Qinliang1   

  1. 1. Affiliated Stomatological Hospital of Zunyi Medical University, Zunyi 563003, China;
    2. Zunyi Medical University, Zunyi 563000, China;
    3. The Fifth Affiliated (Zhuhai) Hospital of Zunyi Medical University, Zhuhai 519180, China
  • Received:2022-05-20 Online:2022-12-28 Published:2022-12-26

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摘要： 目的: 观察低氧预处理人羊膜间充质干细胞(HP-hAMSCs)外泌体对SD大鼠唾液腺放射性功能损伤的修复作用。方法: 试剂盒法分离并鉴定常氧和缺氧预处理人羊膜间充质干细胞(Nor-hAMSCs和HP-hAMSCs)外泌体。电子直线加速器18 Gy一次性照射,建立大鼠唾液腺损伤模型。72只6~8周SD大鼠,每组18只,随机分为Control组、PBS组(照射+PBS组)、NExos组(照射+NExos组)、HExos组(照射+HExos组),采用18 Gy一次性照射建立大鼠唾液腺损伤模型。放射后1 d双侧颌下腺区原位注射,分别于照射后7、14和第28天分别取6只检测大鼠体质量和唾液量,腺体经HE染色进行形态学观察。免疫组织化学染色检测水通道蛋白5(AQP5)的表达。结果: NExos组、HExos组大鼠体重、唾液量均高于PBS组。HExos组唾液量高于NExos组,在第7、28天HExos组大鼠体质量高于NExos组且差异有统计学意义(P<0.05)。颌下腺组织HE染色显示:HExos组的腺体修复能力最强。颌下腺组织AQP5表达的定量分析结果表明:HExos组及NExos组较PBS组在第14、28天光密度值增加(P<0.05),且HExos组高于NExos组,差异有统计学意义(P<0.05)。结论: HP-hAMSCs外泌体能有效修复放射性损伤唾液腺的唾液分泌功能。

关键词: 人羊膜间充质干细胞, 缺氧预处理, 唾液腺, 放射性损伤, 外泌体

Abstract: Objective: To observe the effect of hypoxic pretreatment of human amniotic mesenchymal stem cells (HP-hAMSCs) on the repair of salivary gland radiation function damage in SD rats. Methods: Exosomes of normal oxygen and hypoxia pretreated human amniotic mesenchymal stem cells (Nor-hAMSCs and HP-hAMSCs) were isolated and identified by kit method. Electron linear accelerator 18 Gy one-time irradiation was used to build a model of salivary gland damage in rats. Seventy-two 6-8 weeks SD rats were randomly divided into Control group, PBS group, NExos group, and HExos group, respectively, with 18 in each group. Seven, 14, and 28 days after irradiation, the weight and saliva volume of 6 rats were measured, and the glands were observed. Immunohistochemical staining was used to detect the expression of aquaporin 5 (AQP5). Results: The weight and saliva volume of rats in the NExos group and HExos group were higher than those in the PBS group. The amount of saliva in the HExos group was higher than that in the NExos group, and the weight of rats in the HExos group was higher than that in the NExos group at days 7 and 28 (P<0.05). HE staining of submandibular gland tissue showed that the glands in the HExos group had the strongest repair capacity. Quantitative analysis of AQP5 expression in submandibular glandular tissue showed that the optical density values of HExos group and NExos group increased at day 14 and 28 compared with those of PBS group (P<0.05), and the optical density value of HExos group was higher than that of NExos group (P<0.05). Conclusion: HP-hAMSCs exosomes can effectively repair the salivary secretion function of the salivary glands damaged by radiation.

Key words: human amniotic mesenchymal stem cells, hypoxic preconditioning, salivary glands, radiation damage, exosomes