中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (31): 5017-5022.doi: 10.3969/j.issn.2095-4344.1487

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

深层海水促进糖尿病模型小鼠创面愈合：激活Wnt/β-catenin通路的机制研究

李为明，孙大力，李奕俊，李树民，孙岩波，崔  进，徐鹏远，许青文   

  1. (昆明医科大学第二附属医院胃肠外科二病区，云南省昆明市  650101)
  • 收稿日期:2019-04-27 出版日期:2019-11-08 发布日期:2019-11-08
  • 通讯作者: 许青文，主治医师，昆明医科大学第二附属医院胃肠外科二病区，云南省昆明市 650101
  • 作者简介:李为明，男，1976年生，湖北省麻城市人，汉族，2014年昆明医科大学毕业，博士，副主任医师，主要从事普通外科研究。
  • 基金资助:

    云南省应用基础研究联合专项(2015FB057)，项目负责人：李为明；云南省外科临床营养研究中心子课题(2016NS272)，项目负责人：李为明

Deep seawater promotes cutaneous wound healing in diabetic mice via activating Wnt/β-catenin pathway 

Li Weiming, Sun Dali, Li Yijun, Li Shumin, Sun Yanbo, Cui Jin, Xu Pengyuan, Xu Qingwen   

  1.  (Second Ward of Department of Gastrointestinal Surgery, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunnan Province, China)
  • Received:2019-04-27 Online:2019-11-08 Published:2019-11-08
  • Contact: Xu Qingwen, Attending physician, Second Ward of Department of Gastrointestinal Surgery, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunnan Province, China
  • About author:Li Weiming, MD, Associate chief physician, Second Ward of Department of Gastrointestinal Surgery, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunnan Province, China
  • Supported by:

    the Yunnan Provincial Applied Basic Research Project, No. 2015FB057 (to LWM); the Project of Yunnan Provincial Surgical Clinical Nutrition Research Center, No. 2016NS272 (to LWM)

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摘要：

文章快速阅读：

文题释义：
深层海水：一般指深度超过200 m的循环海水，由于光线无法到达该深度，海水水质稳定而洁净，呈弱碱性，病原菌也极少(深层海水中所含的细菌仅是表层水的千分之一至万分之一)，具有低温、富含矿物质及营养成分等优点，因此应用范围较广泛，可以创造极高的附加值。日本富山、高知、冲绳等县已在海水养殖、康复、美容、饮料食品等方面取得了一系列成果，并在医学领域开展了很多研究。
Wnt信号通路：具有调控皮肤发育和毛囊生长、促进创伤皮肤血管新生及上皮重塑等功能，成为皮肤伤口愈合相关的通路之一。当Wnt通路被激活时，可抑制糖原合成酶3β(GSK-3β)的活性，阻止β-catenin发生磷酸化，进而不被泛素-蛋白酶体系降解，增加细胞内钙离子浓度并激活钙离子敏感信号成分，调节细胞运动和细胞黏着性。
摘要
背景：研究证实深层海水可通过激活AMPK通路下调小鼠对糖的摄入，进而有效缓解高脂肪饮食诱导的小鼠糖尿病进程，然而目前有关深层海水在糖尿病创伤难愈合中的作用研究鲜有报道。
目的：探讨深层海水对糖尿病小鼠创面的修复作用机制。
方法：取健康昆明种雄性小鼠48只(购于昆明医科大学实验动物中心)，通过腹腔注射链脲佐菌素建立糖尿病模型；饲养14 d后建立糖尿病小鼠背部创面模型，随机分4组处理，其中3组分别喂食纯净水(对照组)、自来水(自来水组)、深层海水(深层海水组)，抑制剂组喂食深层海水的同时股静脉注射Wnt/β-catenin抑制剂XAV-939。创面造模后第3，7，10天，检测对照组、自来水组、深层海水组小鼠血糖和体质量、创面愈合及创面组织中β-catenin、GSK-3β和Rspo-3蛋白表达，观察对照组、深层海水组、抑制剂组创面愈合率、创面组织形态变化及血清中丙二醛、超氧化物歧化酶和谷胱甘肽过氧化物酶水平。实验获得中国科学院昆明动物研究所实验动物伦理委员会批准(批准号：KPRC-2017091)。
结果与结论：①深层海水对糖尿病小鼠小鼠体质量和血糖值无影响；②深层海水组不同时间点的创面愈合率明显高于自来水组和对照组(P均< 0.01)；③Western blotting检测显示与对照组和自来水组比较，深层海水组创面组织中β-catenin、Rspo-3蛋白表达升高，GSK-3β蛋白表达下降；④苏木精-伊红染色显示，与对照组比较，深层海水组新生肉芽组织中血管内皮细胞和新生毛细血管增多，炎性细胞浸润较少，成纤维细胞增生明显；抑制剂组与对照组病理表现无明显差异(P > 0.05)；深层海水组不同时间点的创面愈合率明显高于自来水组和抑制剂组(P均<0.01)；⑤与对照组比较，深层海水组超氧化物歧化酶、谷胱甘肽过氧化物酶活性升高(P <0.01，P < 0.001)，丙二醛浓度下降(P < 0.05，P < 0.01)；抑制剂组3指标水平与对照组比较无明显差异(P > 0.05)；⑥结果表明，深层海水通过激活Wnt/β-catenin通路促进糖尿病小鼠创面的愈合。

关键词: 糖尿病, Wnt/β-catenin通路, 深层海水, 创面修复, 自来水, 组织, XAV-939, 小鼠

Abstract:

BACKGROUND: Deep seawater has been shown to effectively alleviate the process of diabetes in mice induced by high fat diet through activating AMPK pathway, but there are few studies about the role of deep seawater in the wound healing of diabetes mellitus.
OBJECTIVE: To explore the mechanism of deep seawater on the cutaneous wound healing in diabetic mice.
METHODS: Forty-eight healthy male Kunming mice (provide by Laboratory Animal Center of Kunming Medical University) were selected and were given intraperitoneal injection of streptozotocin to establish diabetic model. After 14 days of incubation, the back would model was established, and the mouse models were randomly divided into four groups: control group (purified water), tap water group, deep seawater group and inhibitor group (deep seawater + venous injection of Wnt/β-catenin inhibitor, XAV-939). Subsequently, the blood glucose and body mass of model mice were observed at 3, 7 and 10 days after wound modeling, and the healing of the wound was observed. The morphological changes of the wound were evaluated. The expression levels of β-catenin, GSK-3β and Rspo-3, malondialdehyde, superoxide dismutase and glutathione peroxidase in the serum were detected. The study was approved by the Laboratory Animal Ethics Committee of Kunming Institute of Zoology, China Academy of Sciences, approval No. KPRC-2017091.
RESULTS AND CONCLUSION: (1) Deep seawater showed no significant effect on the body mass and blood glucose. (2) The wound healing rate in the deep seawater group was significantly higher than that in the tap water and control groups at different time points (all P < 0.01). (3) Western blot results showed that the expression levels of β-catenin and Rspo-3 were higher in the deep seawater group than the control and tap water groups, but GSK-3β was down-regulated. (4) Hematoxylin-eosin staining showed that compared with the control group, vascular endothelial cells, new capillaries and fibroblast cells proliferation in the deep seawater group were increased, but the number of inflammatory cells got opposite results. However, there was no significant difference between the deep seawater + XAV-939 group and control group (P > 0.05). The healing rate of wound at different time points in the deep seawater group was significantly higher than that in the tap water and control groups (all P < 0.01). (5) Compared with the control group, the expression levels of superoxide dismutase and glutathione peroxidase in serum in the deep seawater group were increased (P < 0.01 and P < 0.001), and malondialdehyde decreased (P < 0.05 and P < 0.01). There was no significant difference between deep seawater + XAV-939 and control groups (P > 0.05). (6) To conclude, deep seawater promotes cutaneous wound healing in diabetic mice via activating Wnt/β-catenin pathway.

Key words: diabetes, Wnt/β-catenin pathway, deep seawater, wound healing, tap water, tissue, XAV-939, mice

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