目的 探究京万红软膏对糖尿病足大鼠的作用及其机制。方法 选取40只健康大鼠，分为对照、模型、京万红软膏、重组人表皮生长因子（rhEGF）4组，每组10只。除对照组外，其余大鼠均建立糖尿病足模型，建模成功后第2天给药，将大鼠足部创面周围用生理盐水清洁后上药，模型组大鼠每日涂抹生理盐水，并与对照组大鼠进行常规喂养；京万红软膏组每天外敷涂抹3 g/kg京万红软膏覆盖创面，rhEGF组每天给予1次rhEGF外用溶液4 000 U。各组均连续给药15 d。通过大鼠足部图像观察创面愈合情况；采用酶联免疫吸附法（ELISA）检测糖基化终末产物（AGEs）、糖基化终末产物受体（RAGE）及血糖水平；苏木精–伊红（HE）染色观察病理学变化情况；免疫印迹检测结缔组织生长因子（CTGF）、细胞外信号调节蛋白激酶（ERK）、p-ERK蛋白表达情况；PCR法检测HMGBA、HMGB1、ERK1、ERK2 mRNA表达情况。结果 与模型组比较，京万红软膏组和rhEGF组大鼠的创面均减小，并以京万红软膏组的效果最为显著（P＜0.05）。与模型组比较，京万红软膏组和rhEGF组大鼠血清中AGEs、RAGE、血糖水平均显著降低（P＜0.05），以京万红软膏组的降低最为显著（P＜0.05）。对照组大鼠皮肤组织结构完整且胶原含量丰富，模型组大鼠表皮结构损伤，有大量炎性及坏死组织；京万红软膏组大鼠表皮生长速度较快，创面组织生长活跃，毛细血管和成纤维细胞数量增多；rhEGF组大鼠组织结构稍显完整，炎性浸出物降低。与模型组比较，京万红软膏组和rhEGF组大鼠创伤组织中CTGF、ERK、p-ERK蛋白表达均明显降低，且以京万红软膏组的降低最为显著（P＜0.05）。与模型组比较，京万红软膏组和rhEGF组大鼠创伤组织中HMGBA、HMGB1、ERK1、ERK2 mRNA表达显著降低，以京万红软膏组的降低最为显著（P＜0.05）。结论 京万红软膏可促进糖尿病足大鼠的创面愈合、减小创伤面积，降低AGEs、RAGE水平，作用机制可能与抑制HMGB及ERK相关通路有关。

Objective To explore the effect and mechanism of Jingwanhong Ointment on diabetic foot rats. Methods Select 40 healthy rats, divided into control, model, Jingwanhong Ointment, and rhEGF groups, each group had 10 rats. The diabetic foot model was established in all rats except the control group. All rats were given the drug on the 2nd day after modeling, and the area around the foot wound was cleaned with normal saline before the drug was applied. Rats in the model group were applied with normal saline every day, and were routinely fed with rats in the control group. Jingwanhong Ointment group applied 3 g/kg Jingwanhong Ointment every day to cover the wound, rhEGF group was given rhEGF external solution 4 000 U once daily. Rats in each group were continuously given drugs for 15 d. The wound healing was observed by foot images of rats. AGEs, RAGE, and blood glucose levels were determined by ELISA. The pathological expression of HE staining was observed. Western blotting was used to detect the expression of CTGF, ERK, and P-ERK proteins. The mRNA expressions of HMGBA, HMGB1, ERK1 and ERK2 were detected. Results Compared with the model control group, the wound surface of Jingwanhong Ointment group and rhEGF group were reduced after treatment, and the effect of Jingwanhong Ointment group was the most significant (P < 0.05). Compared with model group, the expressions of AGEs, RAGE, and blood glucose in serum of Jingwanhong Ointment group and rhEGF group were significantly decreased, and the decrease of AGEs, RAGE and blood glucose in Jingwanhong Ointment group was the most significant (P < 0.05). The skin structure of rats in the control group was intact, and collagen content was rich, while the skin structure of rats in the model group was damaged, and there were a lot of inflammatory and necrotic tissues. In Jingwanhong Ointment group, the epidermis growth rate was faster, wound tissue growth was active, and the number of capillaries and fibroblasts increased. The tissue structure of rhEGF group was slightly intact, and the inflammatory extract decreased. Compared with model group, the protein expressions of CTGF, ERK and p-ERK in Jingwanhong Ointment group and rhEGF group were significantly decreased, and the decrease was most significant in Jingwanhong Ointment group (P < 0.05). Compared with model group, mRNA expressions of HMGBA, HMGB1, ERK1 and ERK2 in Jingwanhong Ointment group and rhEGF group were significantly decreased, and the decrease was most significant in Jingwanhong Ointment group (P < 0.05). Conclusion Jingwanhong Ointment can promote the wound healing, reduce the wound area and the expression levels of AGEs and RAGE in diabetic foot rats, and its mechanism may be related to the inhibition of HMGB and ERK related pathways.

R982

天津市卫健委课题(2015010)