外用溃疡散对糖尿病小鼠溃疡疗效及机制的研究

Study on the Therapeutic Effect and Mechanism of WYK in Diabetic Mice

目的通过制备糖尿病小鼠皮肤溃疡模型,探讨外用溃疡散对糖尿病小鼠皮肤溃疡的治疗效果以及血管生成相关因子VEGF、ERK的表达情况。方法雄性清洁级昆明小鼠30只,随机分为正常对照组(C组),糖尿病对照组(HC组),糖尿病外用溃疡散组(HW组)。HC组和HW组造模成功后,给予HW组外用溃疡散治疗,于小鼠背部涂抹,每日1次;C组和HC组,给予庆大霉素注射液外涂治疗,每日1次;分别于给药第3、6、9日处死小鼠并取材。采用的方法有:蛋白质免疫印记法(Western-blot)、逆转录聚合酶链反应法(RT-PCR)。检测修复过程中血管生成相关因子VEGF、ERK的表达差异。结果VEGF、ERK含量比较:给药3、6、9 d小鼠血清中VEGF含量,HC组明显低于C组(P<0.05);HW组显著高于HC组。血清中ERK含量与VEGF含量基本一致。蛋白印迹检测结果和RT-PCR检测结果基本一致。结论外用溃疡散能够有效促进糖尿病小鼠皮肤溃疡创面的愈合,加快肉芽组织增生,丰富毛细血管和胶原纤维含量,增加微血管含量。外用溃疡散可以通过提高小鼠血清中VEGF、ERK表达水平,且可使其表达该蛋白的峰值提前。

Objective Through the preparation of diabetic mouse skin ulcer model,we investig ated the effect of WYK on the treatment of diabetic skin ulcers and the expressi on of angiogenesis related factorsVEGF、ERK.Methods 30 male clean gradeKunming mice were randomly divided into normal control group(Cgroup),diabetic control group(group HC),diabetic external ulcer powder group(group HW).HC group and HW group after the modeling,HW group was treated with topical ulcer powder treatment,on the back of mice smear,1 times a day;C group and HC group received gentamicin injection with the treatment,1 times a day;the medicine was given 3D,6D,9D mice were killed and drawn.The methods adopted include protein immunoblotting(Western-blot)and reverse transcription polymerase chain reaction(RT-PCR).Differential expression of angiogenesis related factors VEGF and ERK during repair and repair.SPSS 13 software was used to analyze the results of angiogenesis related factors VEGF and ERK.Results VEGF,ERK content comparison:Administration of 3D,6D,9D mice serum VEGF content,HC group was significantly lower than C group(P<0.05);HW group was significantly higher than HC group(P<0.05).Serum ERK content was consistent with VEGF content.The results of Western blot detection and RT-PCR detection are basically the same.Conclusion WYK can effectively promote the healing of diabetic mice skin ulcer wound,accelerate granulation tissue proliferation,rich capillary and collagen fiber content,increase microvascular content.Levels of angiogenesis related factors VEGF,decreased expression of ERK may be the cause of diabetic skin ulcer is difficult to heal the ulcer powder for external use can improve the level of VEGF,the expression of ERK in serum of mice,promote wound healing,and the peak expression of the protein in advance.