目的 探究京萬(wàn)紅軟膏對(duì)糖尿病足大鼠的作用及其機(jī)制。方法 選取40只健康大鼠，分為對(duì)照、模型、京萬(wàn)紅軟膏、重組人表皮生長(zhǎng)因子（rhEGF）4組，每組10只。除對(duì)照組外，其余大鼠均建立糖尿病足模型，建模成功后第2天給藥，將大鼠足部創(chuàng)面周圍用生理鹽水清潔后上藥，模型組大鼠每日涂抹生理鹽水，并與對(duì)照組大鼠進(jìn)行常規(guī)喂養(yǎng)；京萬(wàn)紅軟膏組每天外敷涂抹3 g/kg京萬(wàn)紅軟膏覆蓋創(chuàng)面，rhEGF組每天給予1次rhEGF外用溶液4 000 U。各組均連續(xù)給藥15 d。通過(guò)大鼠足部圖像觀察創(chuàng)面愈合情況；采用酶聯(lián)免疫吸附法（ELISA）檢測(cè)糖基化終末產(chǎn)物（AGEs）、糖基化終末產(chǎn)物受體（RAGE）及血糖水平；蘇木精–伊紅（HE）染色觀察病理學(xué)變化情況；免疫印跡檢測(cè)結(jié)締組織生長(zhǎng)因子（CTGF）、細(xì)胞外信號(hào)調(diào)節(jié)蛋白激酶（ERK）、p-ERK蛋白表達(dá)情況；PCR法檢測(cè)HMGBA、HMGB1、ERK1、ERK2 mRNA表達(dá)情況。結(jié)果 與模型組比較，京萬(wàn)紅軟膏組和rhEGF組大鼠的創(chuàng)面均減小，并以京萬(wàn)紅軟膏組的效果最為顯著（P＜0.05）。與模型組比較，京萬(wàn)紅軟膏組和rhEGF組大鼠血清中AGEs、RAGE、血糖水平均顯著降低（P＜0.05），以京萬(wàn)紅軟膏組的降低最為顯著（P＜0.05）。對(duì)照組大鼠皮膚組織結(jié)構(gòu)完整且膠原含量豐富，模型組大鼠表皮結(jié)構(gòu)損傷，有大量炎性及壞死組織；京萬(wàn)紅軟膏組大鼠表皮生長(zhǎng)速度較快，創(chuàng)面組織生長(zhǎng)活躍，毛細(xì)血管和成纖維細(xì)胞數(shù)量增多；rhEGF組大鼠組織結(jié)構(gòu)稍顯完整，炎性浸出物降低。與模型組比較，京萬(wàn)紅軟膏組和rhEGF組大鼠創(chuàng)傷組織中CTGF、ERK、p-ERK蛋白表達(dá)均明顯降低，且以京萬(wàn)紅軟膏組的降低最為顯著（P＜0.05）。與模型組比較，京萬(wàn)紅軟膏組和rhEGF組大鼠創(chuàng)傷組織中HMGBA、HMGB1、ERK1、ERK2 mRNA表達(dá)顯著降低，以京萬(wàn)紅軟膏組的降低最為顯著（P＜0.05）。結(jié)論 京萬(wàn)紅軟膏可促進(jìn)糖尿病足大鼠的創(chuàng)面愈合、減小創(chuàng)傷面積，降低AGEs、RAGE水平，作用機(jī)制可能與抑制HMGB及ERK相關(guān)通路有關(guān)。

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

天津市衛(wèi)健委課題(2015010)