| Diabetic foot is one of the most common serious complications of diabetes.Its treatment is a hot and difficult point at home and abroad.The rate of amputation and mortality due to diabetic foot is high.It has been reported that the 5-year amputation-free rate of newly diagnosed diabetic foot after treatment is only 50% ~ 60%.Traditional Chinese medicine has accumulated rich clinical experience in the course of treatment,especially the external treatment has great advantages and potential in the field of treating diabetic foot.In this study,a diabetic foot ulcer wound model was established in rats to investigate the effect of she drug "Clematis florida Thunb.var.Plena DE.Don" on wound healing and pathological changes in rats.Wnt signaling pathway was used to study the mechanism of Clematis florida Thunb.var.Plena DE.Don compound prescription in promoting wound healing of diabetic foot ulcer.Clematis florida Thunb.var.Plena DE.Don compound ointment for external use was developed for easy carrying and application.This thesis is divided into three chapters:Chapter one: Effect of Clematis florida Thunb.var.Plena DE.Don compound prescription on wound healing of diabetic foot in ratsSPF male wistar rats were randomly divided into blank control group and Diabetes Group.Diabetic rats were induced by intraperitoneal injection of Streptozocin,and Fasting Blood Glucose ?16.7 mmol/ L was considered as a successful model.The diabetic foot model was established in hyperglycemic rats,and the diabetic foot model was performed in the control rats.Diabetic foot model rats were randomly divided into model control group,positive control group,Clematis florida Thunb.var.Plena DE.Don compound high,medium and low dose group,20 rats in each group,20 rats in blank control group.The wound healing was observed on the 3rd,7th,10 th and 14 th day after treatment.The wound healing rate was measured and calculated.The average wound healing rate of the blank control group was 45.89%,68.89%,80.57% and 94.63% respectively,and that of the model control group was 23.98%,43.90%,56.01% and 75.78% respectively.The healing rate of the Blank Control Group was significantly higher than that of the Model Control Group(p<0.01).The average wound healing rate of the middle dose group was 36.83%,61.95%,73.48% and 91.02%,respectively,and that of the positive control group was 34.74%,54.58%,74.05% and 90.38%,respectively.The healing rate of the Middle Dose Group was significantly higher than that of the Model Control Group(p<0.01),there was no difference with the positive control group.On the 14 th day,except the model control group,the wound healing rate was low,and there was no significant difference among the other groups.The results showed that the wound healing rate of each group increased gradually with the increase of each time point,and the wound healing degree of the Blank Control Group was the best with Normal Blood Glucose,and the treatment effect of the Middle Dose Group was no different from that of the positive control group,can Promote the wound tissue repair.The healing cycle of diabetic foot ulcers in rats was about 14 days.The second chapter : Study on the mechanism of Clematis florida Thunb.var.Plena DE.Don compound prescription in the treatment of diabetic foot in ratsThe rats were randomly divided into 3 groups at 3 days,7 days and 10 days,and 5 rats were killed after collecting ulcer wound tissue.The tissue samples were stained with HE.Western blot was used to detect the protein expression levels of ?-catenin and glycogen synthase kinase-3? The results show that:1.HE staining observation: compared with the blank control group,the Model Control Group had the longest duration of wound inflammation,the Middle Dose Group had the best effect in the three dose groups,and the fibroblasts and new blood vessels appeared earlier and more frequently,granulation tissue matured faster than the model control group,and there was no difference in wound healing between the two groups.2.?-catenin and GSK-3? protein expression: on the 3rd,7th and 10 th day after Administration,?-catenin protein expression was lower and GSK-3 protein expression was higher in model control group than that in Blank Control Group(p<0.05),the expression level of ?-catenin protein in medium dose group was higher than that in Model Control Group(p<0.05),and the expression level of GSK-3? protein was lower than that in Model Control Group(p<0.05).Twelve hour compound can increase the expression of ?-catenin protein and decrease the expression of GSK-3? protein.The mechanism of twelve hour compound on diabetic foot is related to the downstream proteins of Wnt signal pathway ?-catenin and GSK-3?.The third chapter,Clematis florida Thunb.var.Plena DE.Don compound ointment preparation technology research.To compare the stability of different matrix formulations of ointment,the best matrix formulation was selected for single factor investigation,on the basis of which the Box-Behnken design was used to select the best matrix formulation,and the different ratio of drug to matrix was used to prepare the ointment,to determine the optimum content of Clematis florida Thunb.var.Plena DE.Don compound ointment and observe its appearance.The results showed that the optimum matrix formulation was O/W Matrix with polysorbate as emulsifier,and the optimum Matrix ratio was glycerol monostearate: tween-80: White Vaseline(0.8:4:3).To sum up,Clematis florida Thunb.var.Plena DE.Don compound prescription can effectively promote the wound healing of diabetic foot ulcer in rats,of which the Middle Dose Group has the best effect.The mechanism of promoting the healing of diabetic foot in rats is related to inhibiting Wnt pathway,which can increase the expression of ?-catenin protein and decrease the expression of GSK-3? protein.The prescription of Clematis florida Thunb.var.Plena DE.Don compound ointment is white vaseline 3 g,Monostearin 0.8 g,Lanolin 0.9 g,Stearin 1g,tween-80 4 g,Glycerin 3 g,distilled water 16 g,Sodium lauryl sulfate 0.5g. |
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