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| 人脐带间充质干细胞来源外泌体对小鼠压疮的治疗作用及机制 |
| 罗雅婷1,解婧2,许涛3,谢水林4,张坚松1*,刘劼5* |
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(1. 湖南师范大学医学院基础医学系, 长沙 410013;
2. 复旦大学附属肿瘤医院中西医结合科, 上海 200032;
3. 上海交通大学附属第六人民医院麻醉科, 上海 200233;
4. 华南理工大学生物科学与工程学院, 广州 510720;
5. 云南省第一人民医院转化医学中心, 昆明 650034
*通信作者) |
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| 摘要: |
| 目的 探索人脐带间充质干细胞（hucMSC）来源外泌体（hucMSC-Exo）对小鼠4期压疮创面愈合的作用及其机制。方法 原代分离、培养人脐带间充质干细胞，取第5代（P5代）细胞培养上清，通过差速离心法提取hucMSC-Exo。将27只成功制备4期压疮模型的BALB/c小鼠随机分为hucMSC-Exo治疗组（创周皮下注射100 μg溶解于100 μL PBS中的hucMSC-Exo）、PBS对照组（创周皮下注射100 μL PBS）及模型对照组（未给予hucMSC-Exo和PBS处理）。治疗后评估压疮创面愈合情况，于0、3、7、10、14 d拍照并计算创面愈合率；取治疗后7、14 d皮肤样本行H-E染色和Masson染色观察创面病理变化；取治疗后7 d皮肤样本行免疫组织化学染色检测与瘢痕形成相关的α平滑肌肌动蛋白（α-SMA）表达；取治疗后7、14 d血清检测氧化应激指标超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量的动态变化。结果 hucMSC-Exo治疗组较PBS对照组及模型对照组创面愈合速度加快，瘢痕形成减少，创面愈合率升高。组织病理学结果显示，与PBS对照组及模型对照组比较，hucMSC-Exo治疗组创面损伤程度、炎性浸润情况明显减轻，胶原沉积量增多，胶原纤维排列分布有序。免疫组织化学染色结果显示，hucMSC-Exo治疗组α-SMA蛋白表达少于PBS对照组及模型对照组。血清生物化学检测结果示，hucMSC-Exo治疗组SOD活性在治疗后7 d高于PBS对照组及模型对照组（P均<0.05），到14 d接近正常水平； MDA含量在7 d和14 d均低于PBS对照组及模型对照组（P均<0.05）。结论 皮下注射hucMSC-Exo可促进小鼠4期压疮创面愈合，减少瘢痕形成，其机制可能与hucMSC-Exo改善体内氧化应激水平有关。 |
| 关键词: 脐带间充质干细胞 外泌体 压疮 伤口愈合 氧化性应激 |
| DOI：10.16781/j.CN31-2187/R.20220373 |
| 投稿时间：2022-04-03修订日期：2022-05-07 |
| 基金项目:国家自然科学基金地区科学基金（82060242）. |
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| Therapeutic effect and mechanism of human umbilical cord mesenchymal stem cell-derived exosome on wound healing of pressure ulcers in mice |
| LUO Ya-ting1,XIE Jing2,XU Tao3,XIE Shui-lin4,ZHANG Jian-song1*,LIU Jie5* |
(1. Department of Basic Medicine, Medical College of Hunan Normal University, Changsha 410013, Hunan, China;
2. Department of TCM-WM Integrated Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, China;
3. Department of Anesthesiology, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China;
4. School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510720, Guangdong, China;
5. Center for Translational Medicine, The First People's Hospital of Yunnan Province, Kunming 650034, Yunnan, China
*Corresponding authors) |
| Abstract: |
| Objective To explore the effect and mechanism of human umbilical cord mesenchymal stem cell (hucMSC)-derived exosome (hucMSC-Exo) on wound healing of stage 4 pressure ulcers in mice. Methods hucMSCs were isolated and cultured in primary culture. hucMSC-Exos were extracted from the supernatant of hucMSCs at passage 5 (P5) through differential centrifugation. Twenty-seven BALB/c mice with stage 4 pressure ulcers were randomly divided into hucMSC-Exo group (injected subcutaneously with 100 μg hucMSC-Exo dissolved in 100 μL phosphate-buffered saline[PBS]), PBS group (injected subcutaneously with 100 μL PBS), and model group (without injection of hucMSC-Exo or PBS). The wound healing of pressure ulcers was assessed after treatment, and the wound healing rate was estimated on day 0, 3, 7, 10, and 14. Hematoxylin-eosin (H-E) staining and Masson staining were used to observe the pathological changes of the wound on day 7 and 14 after treatment. The expression of α-smooth muscle actin (α-SMA) protein related to scar formation was detected by immunohistochemical staining with skin samples on day 7 after treatment. Serum samples were obtained on day 7 and 14 after treatment to detect the dynamic changes of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. Results Compared with the PBS and model groups, the hucMSC-Exo group had much shorter wound healing time, less scar formation, and higher wound healing rate. Histopathology examination showed that, compared with the PBS and model groups, the wound injury and inflammatory infiltration in the hucMSC-Exo group were significantly decreased, collagen deposition was greatly boosted, and collagen fibers were orderly arranged and distributed. Immunohistochemical staining showed that the expression of α-SMA protein in the hucMSC-Exo group was lower than those in the PBS and model groups. The results of serum biochemical test showed that SOD activity in the hucMSC-Exo group was higher than those in the PBS and model groups (both P<0.05) on day 7 after treatment, approaching the normal level on day 14; MDA content was lower in the hucMSC-Exo group than the PBS and model groups on day 7 and 14 (all P<0.05). Conclusion Subcutaneous injection of hucMSC-Exo can promote wound healing and minimize scar formation in mice with stage 4 pressure ulcers. The mechanism may be related to the improvement of in vivo oxidative stress levels by hucMSC-Exo. |
| Key words: umbilical cord mesenchymal stem cells exosome pressure ulcer wound healing oxidative stress |
