| Objective: 1.To arrange the medicine and summarize the law of diabetic ulcer patients. 2.To explore the mechanism of refractory diabetic ulcer and promotion healing of cooling-blood and eliminating-stasis from the Wnt signaling pathway by establishing the rat models.Methods: 1.Collecting the information of 171 diabetic ulcer patients, who were enrolled according to the inclusion criteria, and recording the syndrome and drugs. Then,the law of diabetic ulcer was explored by analyzing the categories, resistance, flavor,meridian tropism of traditional Chinese medicine, and Chinese patent medicine of oral,external and intravenous drip. 2.Diabetes mellitus(DM) animal models were established in the female Wistar rats by intraperitoneal injection of low-dose Streptozotocin following high-fat diet feeding. A circular wound was made on the dorsum of the rats in both control group and DM group. And then, the ulcer rats were classified into 5 groups: the control ulcer group(CU group, the rats were treated by change dressing with normal saline gauze and 0.9%NS ig), the diabetic ulcer group(DU group, the rats were treated by change dressing with normal saline gauze and 0.9%NS ig), the diabetic ulcer with Yu-yang-ling Ointment group(DU-Yu group, the rats were treated by change dressing with Yu-yang-ling Ointment and 0.9%NS ig), the diabetic ulcer with Simiao Yong'an group(DU-Simiao group, the rats were treated by change dressing with normal saline gauze andSimiao Yong'an Decoction ig), the diabetic ulcer with Simiao Yong'an and Yu-yang-ling Ointment group(DU-Simiao-Yu group, the rats were treated by change dressing with Yu-yang-ling Ointment and Simiao Yong'an Decoction ig). The condition of wound healing was recorded and the changes of structure of the wound tissues were observed by HE staining in the 5 groups at 3rd, 7th and 14 th d after wounding. Also, the protein and m RNA expression of ?-catenin, GSK-3?, Rspo-3 in wound tissues were detected by ELISA and RT-PCR.Results: 1.The main categories of TCM are heat-clearing drugs, promotingcirculation drugs, tonic drugs, and eliminating-dampness drugs. And their resistance were mostly cool, their flavor were mostly bitter, sweet and pungent, and their meridian tropism were mostly liver, spleen, heart, and stomach. The blood-cooling and circulation-promoting drugs were the most frequently used in oral herbs and external patent medicine, while the blood-activating and stasis-eliminating drugs in oral and intravenous drip patent medicine. Also, the patients in damp-heat syndrome are in accordance with this law. 2.(1)Compared with the control group, there was significant difference in the food intake, water intake, excretion volume, body weight and blood glucose in the DM group after STZ(P<0.05).(2) The wound healing rate of treatment group at each time point were all higher than DU group, but lower than CU group.(3)The?-catenin protein expression in serum in CU group> DU-Simiao-Yu group> DU-Simiao group> DU-Yu group> DU group at 3rd, 7th and 14 th d after wounding, while the ranking of GSK-3? in all groups were opposite. And the Rspo-3 protein was undetectable in serum.(4)The protein and m RNA expression of ?-catenin, Rspo-3 in wound tissues in treatment group were higher than DU group and lower than CU group. Although the expression in the treatment group were not identical at different time point, the DU-Simiao-Yu was all the highest in the 3 groups. The protein and m RNA expression of GSK-3? in wound tissues in treatment group were lower than DU group and higher than CU group, and the DU-Simiao-Yu was all the lowest in the 3 treatment groups.Conclusion: 1.The principle of treatment in diabetic ulcer is cooling-blood and eliminating-stasis. 2.The down-regulation of the Wnt/?-catenin, probably resulting fromthe decreased level of Rspo, may be one of the reasons delaying the diabetic ulcer healing.3.Simiao Yong'an Decoction and Yu-yang-ling Ointment can promote the healing of diabetic ulcer by regulating Wnt/?-catenin signaling pathway, but the mechanism of the two different therapeutic methods are different, and combination them maybe better. 4.The therapy of cooling-blood and eliminating-stasis can be effective method to diabeticulcer. |
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