中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (5): 724-729.doi: 10.12307/2022.118

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

深层海水通过激活PI3K/Akt通路促进糖尿病小鼠创伤的愈合

李为明，许青文，李奕俊，孙岩波，崔  进，徐鹏远   

  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:2020-09-30 修回日期:2020-10-13 接受日期:2021-01-19 出版日期:2022-02-18 发布日期:2021-10-28
  • 通讯作者: 李为明，副主任医师，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(昆医联合专项)资助(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资助(2016NS272)，项目负责人：李为明；云南省卫健委高层次人才项目资助(D-2017037)，项目负责人：李为明

Deep seawater promotes wound healing in diabetic mice by activating PI3K/Akt pathway

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

  1. Second Ward of Department of Gastrointestinal Surgery, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunnan Province, China
  • Received:2020-09-30 Revised:2020-10-13 Accepted:2021-01-19 Online:2022-02-18 Published:2021-10-28
  • Contact: Li Weiming, 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:
    Yunnan Provincial Applied Basic Research Project, No. 2015FB057 (to LWM); the Subproject of Yunnan Provincial Surgical Clinical Nutrition Research Center, No. 2016NS272 (to LWM); High-level Talent Project of Yunnan Provincial Health and Construction Commission, No.D-2017037 (to LWM) 

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摘要：

文题释义：
深层海水：是指水深200 m以下的海水，水质纯净而稳定，呈弱碱性，富含人体细胞所需的无机盐和微量元素，其水分子团与人体水分、血液结合角极其近似，极易被人体吸收。


背景：前期研究证实，深层海水不仅影响小鼠的耐受和免疫，还对创面修复具有重要的调控作用。
目的：探讨深层海水修复糖尿病小鼠创面过程中对PI3K/Akt通路的影响。
方法：取48只昆明种小鼠，建立糖尿病模型，造模成功后在背部制作1 cm2的创面，随机分4组处理，分别饮用纯净水(对照)、自来水、深层海水及深层海水+股静脉注射PI3K/Akt通路抑制剂LY294002，每组12只。创面造模后第3，7，10天，观察创面愈合情况，Western blot检测创面组织中血管内皮生长因子、碱性成纤维细胞生长因子、缺氧诱导因子1α、p-PI3K、p-Akt、PI3K和Akt表达水平，qRT-PCR和Western blot检测胶原Ⅰ、胶原Ⅲ、白细胞介素1β、肿瘤坏死因子α和转化生长因子β1的表达水平；创面造模后第3，10天，苏木精-伊红染色观察创面形态变化。实验获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深层海水组各时间点的创面愈合率高于对照组、自来水组(P < 0.01)，深层海水+LY294002组与对照组比较差异无显著性意义(P > 0.05)；②苏木精-伊红染色显示，对照组、自来水组、深层海水+LY294002组第3天创面有大量炎性细胞浸润及少量纤维母细胞，第10天可见肉芽组织及毛细血管和纤维母细胞；深层海水组第3天可见纤维母细胞、毛细血管和少量炎性细胞，第10天可见新生肉芽组织及较多胶原纤维和少量毛细血管；③Western blot检测显示，深层海水组各时间点的血管内皮生长因子、碱性成纤维细胞生长因子、缺氧诱导因子1α、p-PI3K和p-Akt蛋白表达均高于对照组、自来水组(P < 0.01)；④qRT-PCR和Western blot检测显示，深层海水组各时间点的胶原Ⅰ、胶原Ⅲ和转化生长因子β1表达均高于对照组、深层海水+LY294002组(P < 0.05或P < 0.01)，白细胞介素1β、肿瘤坏死因子α表达均低于对照组、深层海水+LY294002组(P < 0.05或P < 0.01)；⑤结果表明，深层海水通过激活PI3K/Akt通路促进糖尿病小鼠创面愈合。
https://orcid.org/0000-0001-9220-514X (李为明) 

关键词: 糖尿病, PI3K/Akt通路, 深层海水, 创面愈合, 实验动物

Abstract: BACKGROUND: Preliminary studies have confirmed that deep seawater not only affects the tolerance and immunity of mice, but also has an important regulatory effect on wound repair.
OBJECTIVE: To explore the effect of deep seawater on the PI3K/Akt pathway in the process of wound repair in diabetic mice.
METHODS: Forty-eight Kunming mice were enrolled to establish a diabetes model. A 1 cm2 wound was made on the back after successful modeling, and all the mouse models were randomized into four groups for corresponding treatments, including drinking purified water (control), tap water, deep seawater and deep seawater+LY294002, with 12 rats in each group. On the 3rd, 7th, and 10th days after wound modeling, the wound healing was observed, and western blot was used to detect the expression levels of vascular endothelial growth factor, basic fibroblast growth factor, hypoxia-inducible factor 1α, p-PI3K, p-Akt, PI3K and Akt in the wound tissue. qRT-PCR and western blot were used to detect the expression levels of collagen type I, collagen type III, interleukin-1β, tumor necrosis factor-α and transforming growth factor-β1 mRNA. On the 3rd and 10th days after wound modeling, hematoxylin-eosin staining was used to observe the morphological changes of the wound. The study protocol was approved by the Animal Ethic Committee of Kunming Institute of Zoology, Chinese Academy of Science with an approval No. KPRC-2017091.
RESULTS AND CONCLUSION: The wound healing rate of the deep seawater group was higher than that of the control group and the tap water group (P < 0.01), while there was no significant difference in the wound healing rate between the tap water group and deep seawater+LY294002 group (P > 0.05). Hematoxylin-eosin staining showed that there were a large number of inflammatory cells infiltrated and a few fibroblasts on the wound surface in the control group, tap water group, and deep seawater+LY294002 group on the 3rd day, and granulation tissue, capillaries and fibroblasts were visible on the 10th day. In the deep seawater group, fibroblasts, capillaries and a few inflammatory cells were seen on the 3rd day, and new granulation tissue, more collagen fibers and a few capillaries were seen on the 10th day. Western blot analysis showed that the expressions of vascular endothelial growth factor, basic fibroblast growth factor, hypoxia-inducible factor 1α, p-PI3K and p-Akt in the deep seawater group were higher than those in the control group and the tap water group on each observational time point (P < 0.01). qRT-PCR and western blot results showed that the expressions of collagen type I, collagen type III and transforming growth factor-β1 in the deep seawater group were higher than those in the control group and deep seawater+LY294002 group on each observational time point (P < 0.05 or P < 0.01), while the expression levels of interleukin-1β and tumor necrosis factor-α were lower than those in the control group and deep seawater+LY294002 group (P < 0.05 or  P < 0.01). To conclude, deep seawater can promote wound healing in diabetic mice via activating the PI3K/Akt pathway.

Key words: diabetics mellitus, PI3K/Akt pathway, deep seawater, wound healing, experimental animal

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