Department of Clinical and Experimental Medicine, Section of Pharmacology, Medical School, University of Messina, Messina, Italy.
Department of Human Pathology, University of Messina, Messina, Italy.
Br J Pharmacol. 2018 Feb;175(4):644-655. doi: 10.1111/bph.14113. Epub 2018 Jan 18.
Diabetic patients with non-healing ulcers have a reduced expression of VEGF. Genetically diabetic mice have an altered expression pattern of VEGF and its receptor, VEGF receptor 2 (VEGFR-2). In diabetic wounds, the microRNAs, miR15b and miR200b, which respectively inhibit VEGF and VEGF-R2 mRNAs, are up-regulated, further affecting the impaired angiogenesis. We investigated whether anti-miRs directed toward miR15b and miR200b could improve wound repair in genetically diabetic mice.
Skin wounds were produced on the backs of female diabetic mice. The anti-miRs (antimiR15b, antimiR200b or antimiR15b/200b) at 10 mg·kg , or vehicle were applied to the wound edge. Mice were killed on days 7, 14 and at time of complete wound closure. Levels of mRNA and protein of angiogenic mediators and their receptors were measured with RT-qPCR and Western blotting. Wounds were examined by histological and immunochemical methods.
mRNA expression of VEGF, VEGFR-2, angiopoietin-1 and its receptor TEK were evaluated after 7 and 14 days. Protein levels of VEGF and transglutaminase II were measured at day 7, while VEGFR-2 and Angiopoietin-1 were measured at day 14. Histological features and the time to achieve a complete wound closure were also examined. Treatment with the anti-miRs improved the analysed parameters and the co-treatment resulted the most effective.
The results suggest that the inhibition of miR15b and miR200b may have a potential application in diabetes-related wound disorders.
患有难愈性溃疡的糖尿病患者 VEGF 表达降低。遗传型糖尿病小鼠的 VEGF 及其受体 VEGF 受体 2（VEGFR-2）表达模式发生改变。在糖尿病伤口中，分别抑制 VEGF 和 VEGF-R2 mRNA 的 microRNAs，miR15b 和 miR200b，上调，进一步影响受损的血管生成。我们研究了针对 miR15b 和 miR200b 的抗-miR 是否可以改善遗传型糖尿病小鼠的伤口修复。
在雌性糖尿病小鼠的背部产生皮肤伤口。将抗-miR（antimiR15b、antimiR200b 或 antimiR15b/200b）以 10mg·kg 的剂量施用于伤口边缘。在第 7、14 天和完全伤口闭合时杀死小鼠。使用 RT-qPCR 和 Western blotting 测量血管生成介质及其受体的 mRNA 和蛋白水平。通过组织学和免疫化学方法检查伤口。
在第 7 和第 14 天评估了 VEGF、VEGFR-2、血管生成素-1 及其受体 TEK 的 mRNA 表达。在第 7 天测量了 VEGF 和转谷氨酰胺酶 II 的蛋白水平，而在第 14 天测量了 VEGFR-2 和血管生成素-1 的蛋白水平。还检查了组织学特征和实现完全伤口闭合的时间。用抗-miR 治疗改善了分析参数，并且联合治疗最有效。
结果表明，抑制 miR15b 和 miR200b 可能在与糖尿病相关的伤口疾病中有潜在的应用。
