愈疡灵软膏对糖尿病溃疡大鼠Wnt/β-catenin信号通路的影响

Impact of Yuyangling Ointment (愈疡灵软膏) on Wnt/β-catenin Signal Pathway of Rats with Diabetic Ulcer

目的观察愈疡灵软膏对糖尿病溃疡大鼠创面愈合的影响及其可能作用机制。方法 60只Wistar大鼠随机分为对照组、模型组、凡士林组、愈疡灵组,每组15只。除对照组外其余各组采用高脂高糖饲料联合小剂量链脲佐菌素液腹腔注射法制备糖尿病大鼠模型,后各组大鼠均制作溃疡模型。于溃疡造模后第2天愈疡灵组大鼠创面以愈疡灵软膏涂敷其上,约厚2 mm,外用无菌纱布包扎固定;凡士林组采用凡士林油纱外敷;对照组及模型组采用生理盐水纱布外敷,各组每日换药1次。观察给药第3、7、14天各组大鼠创面愈合情况,并采用HE染色法检测创面组织形态结构的变化,检测溃疡组织中Wnt信号通路相关的β-连环素(β-catenin)、糖原合成激酶-3β(GSK-3β)、R-脊椎蛋白3(Rspo-3)蛋白及mRNA的变化。结果与对照组比较,模型组、凡士林组、愈疡灵组大鼠在第3、7、14天创面愈合率,创面组织中β-catenin、Rspo-3蛋白及mRNA表达均降低,GSK-3β蛋白及mRNA表达均升高(P<0.05)。与模型组、凡士林组比较,愈疡灵组大鼠的创面愈合率、创面组织病理学变化、创面组织中β-catenin、GSK-3β、Rspo-3蛋白及mRNA的表达水平均得到不同程度改善,尤其以第14天最为明显(P<0.05)。结论愈疡灵软膏可促进糖尿病溃疡大鼠创面的愈合,其机制可能通过对Wnt/β-catenin信号通路进行调节。

Objective To observe the impact and possible mechanism of Yuyangling Ointment on wound healing in rats with diabetic ulcer. Methods Sixty Wistar rats were randomized into a control group,a model group,a vaseline group and a Yuyangling group,with 15 rats in each group. Rats were given high-sugar-fat-diet and small dose streptozotocin intraperitoneal injection to establish diabetes rat models except those in the control group. Rats in all groups were established ulcer models. The second day after ulcer modeling,the wound surface of rats in the Yuyangling group was plastered Yuyangling Ointment,with the thickness of about 2 mm,bandaged and fixed externally by sterile gauze. The vaseline group applied vaseline oil gauze externally. The control group and the model group applied normal saline gauze externally. Each group changed dressing once every day. The wound healing of rats in all groups was examined on the 3rd,7th and 14 th day of administration. The changes of wound tissue morphological structure were detected with hematoxylin and eosin（ HE） stain. The changes of β-catenin,glycogen synthase kinase-3β（ GSK-3β）,R-spinal protein 3（ Rspo-3） and mRNA in ulcer tissue,related to Wnt signal pathway,were detected.Results Compared with the control group,the wound healing rate on the 3rd,7th and 14 th day,as well as the expressions of β-catenin,Rspo-3 and mRNA decreased,but the expressions of GSK-3βand mRNA increased in the model group,the vaseline group,and the Yuyangling group（ P 〈 0. 05）. Compared with the model group and the vaseline group,the wound healing rate,the pathological changes of wound surface tissue,as well as the expressions ofβ-catenin,GSK-3β,Rspo-3 and mRNA were improved by different degree in Yuyangling group. The changes on the14 th day were the most obvious（ P 〈 0. 05）. Conclusion Yuyangling Ointment might improve wound healing of rats with diabetic ulcer. The mechanism might be regulated with the signal pathway of Wnt / β-catenin.