上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

局部应用胰岛素调控基质金属蛋白酶对糖尿病创面血管新生的影响

常清璇,杨沛瑯,刘琰,章雄   

  1. 上海交通大学 医学院附属瑞金医院烧伤整形科， 上海 200025
  • 出版日期:2015-09-28 发布日期:2015-09-30
  • 通讯作者: 章雄, 电子信箱: xiong@medmail.com.cn。
  • 作者简介:常清璇（1986—）， 女， 硕士生; 电子信箱: 604811511@qq.com。
  • 基金资助:

    国家自然科学基金（81170761， 81270909）

Effect of regulating expression of matrix metalloproteinase by topical application of insulin on angiogenesis of diabetic wounds

CHANG Qing-xuan, YANG Pei-lang, LIU Yan, ZHANG Xiong   

  1. Department of Burn and Plastic Surgery， Ruijin Hospital， Shanghai Jiao Tong University School of Medical, Shanghai 200025, China
  • Online:2015-09-28 Published:2015-09-30
  • Supported by:

    National Natural Science Foundation of China, 81170761， 81270909

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摘要：

目的  研究局部应用胰岛素对血管内皮生长因子（Vegf）及基质金属蛋白酶2/9（Mmp-2/9）的调控，探讨两者在促进糖尿病创面血管新生的作用及相关性。方法  60只Wistar大鼠经高脂饮食喂养和小剂量链脲佐菌素(STZ) 多次注射诱导糖尿病。5 周后，在大鼠背部制造4个直径为0.9 cm的全厚皮肤缺损创面，每天以对角型在两个创面滴加20 μL生理盐水（生理盐水组）和0.1 U胰岛素/20 μL生理盐水（胰岛素组），直至创面完全愈合。观察并记录创面愈合速度，于第3、7、13、21日取材。H-E染色观察愈合后创面皮肤组织结构，7、13 d创面血管新生情况；real-time PCR方法检测Vegf、Vegfr-2、Mmp-2/9的mRNA表达水平。结果  与生理盐水组相比，胰岛素组创面愈合时间明显缩短，组间差异均有统计学意义（P<0.05）。21 d创面愈合后胰岛素组与生理盐水组相比表皮钉突数量增多，真皮组织致密，第7、13日新生血管数量明显增多。与生理盐水组相比，胰岛素组Vegf、Vegfr-2、Mmp-2/9 mRNA水平在第3、7、13日呈逐渐递增状态，13 d到达峰值，随后下降。第21日Vegf、Vegfr-2 mRNA表达仍高于对照组，而Mmp-2/9 mRNA相比生理盐水组表达下降。组间差异具有统计学意义（P<0.05）。结论  局部应用胰岛素促进Vegf、Vegfr-2 mRNA的表达上调，调控Mmp-2/9 mRNA的表达，从而促进糖尿病创面的血管新生。

关键词: 胰岛素, 糖尿病, 血管新生, Vegf, Mmp-2, Mmp-9

Abstract:

Objective  To investigate the regulation of vascular endothelial growth factor (Vegf) and matrix metalloproteinase 2/9 (Mmp-2/9) by topical application of insulin and explore the effects of Vegf and Mmp-2/9 on promoting the angiogenesis of diabetic wounds and their correlation. Methods  The diabetes of 60 Wistar rats were induced by multiple intraperitoneal injections with a low dose of streptozocin (STZ) after high-fat diet feeding. After 5 weeks, four full-thickness skin wounds with the diameter of 0.9 cm were made on the back of rats. Two wounds were treated with 20 μL saline (saline group) and insulin of 0.1 U/20 μL saline (insulin group) in diagonal form each day until complete healing. The healing speed of wounds was observed and recorded and samples were harvested on 3, 7, 13, and 21 d. The structure of skin tissues of healed wounds and angiogenesis of wounds on 7 and 13 d were observed by H-E staining. The mRNA expressions of Vegf, Vegfr-2, and Mmp-2/9 were detected by real-time PCR. Results  Compared with the saline group, the healing speed of the insulin group was faster and the difference between two groups was statistically significant (P<0.05). The number of spikes in the epidermis of insulin group was larger than that of the saline group on 21 d after wounds were healed and dermal tissues were denser. The number of angiogenic blood vessel of the insulin group was larger than that of the saline group on 7 and 13 d. The mRNA expressions of Vegf, Vegfr-2, and Mmp-2/9 of the insulin group increased progressively on 3, 7, and 13 d, reached peak on 13 d, and then decreased. The mRNA expressions of Vegf and Vegfr-2 were still higher than those of the saline group on 21 d and the  mRNA expression of Mmp-2/9 decreased. The difference between two groups was statistically significant (P<0.05). Conclusion  Topical application of insulin promotes the up-regulation of mRNA expressions of Vegf and Vegfr-2, regulates the mRNA expression of Mmp-2/9, and promotes the angiogenesis of diabetic wounds.

Key words: insulin, diabetes, angiogenesis, Vegf, Mmp-2, Mmp-9