温阳生肌膏对糖尿病大鼠阴证慢性创面CD34和VEGF表达及血管重建的影响

Effect of Wenyang Shengji ointment on(温阳生肌膏)the expressions of CD34 and VEGF in chronic wound and vascular reconstruction of diabetic rats with Yin Syndrome(阴证)

目的:探讨温阳生肌膏对糖尿病大鼠阴证慢性创面的治疗机制。方法:60只雄性SD大鼠采用高脂乳剂灌胃+腹腔注射链脲佐菌素的方法制备2型糖尿病大鼠模型,再以股部外侧注射氢化可的松及用打孔器在背部两侧制备全层皮肤缺损的方法制备阴证创面模型。将模型大鼠随机分为模型组、实验组和对照组(n=20)。模型组不做处理,其余2组分别给予温阳生肌膏和象皮生肌膏外敷,每日1次,共21 d。观察造模后3、7、14、21 d创面愈合大体情况和造模后14 d的创面组织病理学变化;免疫组化检测造模后7和14 d创面组织CD34和血管内皮生长因子(VEGF)的表达;测定造模后3、7、14 d微血管密度(MVD)的变化。结果:实验组大鼠创面愈合率明显高于模型组及对照组(P<0.05);造模后3~7 d,实验组皮损处新生肉芽组织MVD较模型组及对照组显著升高(P<0.05);造模后14 d,实验组皮损处新生肉芽组织CD14和VEGF水平与模型组及对照组相比较显著升高(P<0.05);实验组肉芽组织生长旺盛,毛细血管丰富,炎症细胞较少。结论:温阳生肌膏可促进糖尿病大鼠阴证创面的愈合,其作用机制可能与增加局部皮损处新生肉芽组织中的新生微血管数量有关。

Objective:To explore the therapeutic mechanism of Wenyang Shengji ointment(温阳生肌膏)for treatment of chronic wound in diabetic rats with Yin syndrome.Methods:The type 2 diabetes rats model was established with 60 male SD rats by intragastric administration of high fat emulsion and intraperitoneal injection of streptozotocin,and the Yin syndrome wound model was made by lateral femoral injection of hydrocortisone and the full thickness skin defects prepared with punching in both sides of the back.The model rats were randomly divided into model group,experimental group and control group(20 each).Rats in model group were not treated,and in the other two groups were given external application of Wenyang Shengji ointment and Xiangpi Shengji ointment,once a day for 21 days.At 3rd,7th,14th and 21st day after intervention,rats in each group were observed for wound healing and histopathologic changes of local wound tissues.The expressions of CD34 and vascular endothelial growth factor(VEGF)were detected by immunohistochemistry.The changes of microvessel density(MVD)were observed at 3rd,7th and 14th day after intervention.Results:The wound healing rate of rats was significantly higher in experimental group than in model group and control group(P<0.05).After 3 to7 days of administration,the MVD of new granulation tissue in the skin lesion increased significantly in experimental group compared with that in model group and control group(P<0.05).Fourteen days after administration,CD14 and the VEGF level in new granulation tissue of the skin lesions increased significantly in experimental group compared with that in model group and control group(P<0.05).In the experimental group,the granulation tissue grew vigorously with abundant capillaries and less inflammatory cells.Conclusions:Wenyang Shengji ointment can promote the healing of wound in diabetic rats with Yin syndrome,and the mechanism may be related to the number increase of new microvessels in the new granulation tissue of local lesions.