| AIM: In this study,diabetic mouse skin ulcer model was taken as the research object to observe the effects of MEB on the skin healing of the model mice,and by detecting the apoptosis of skin granulation tissue cells and the protein expression levels of Fas,Fas L,caspase3 and caspase8 in the tissue,To explore the mechanism of MEBO on the healing of skin ulcer wound in diabetic mice and to provide experimental basis for the clinical treatment of diabetic skin ulcer.Materials and Methods: A total of 40 SPF male mice of Kunming species were randomly divided into 4 groups: blank control group,model control group,Kangfuxin liquid group and MEBO group,with 10 mice in each group.On the first day before the experiment,mice were fed for 24 hours without food.Except for blank control group,the diabetic mouse model was induced by intritoneal injection of streptozotocin(STZ)on the second day.After 72 hours of STZ injection,blood glucose was randomly selected for testing for 3 times.The hair was removed in the middle dorsal region of all mice,and a scalpel was used to make an incision about L.5cm long,and the depth of the incision was to expose the granulation layer.The wound was stimulated with 50% glacial acetic acid to make it conform to the characteristics of diabetic skin ulcer.Mice in the blank control group and model control group were treated with sterile normal saline on the wound surface,and Kangfuxin liquid and MEBO were selected for external intervention on the ulcer surface in the Kangfuxin liquid group and the MEBO group,once a day,for consecutive 7 days.The healing of ulcer surface was observed after intervention in each group.Granulation tissue of the wound was collected,and HE staining sections were made for histopathological observation.The apoptotic changes of granulation tissue cells were observed by TUNEL assay.The protein expression levels of Fas,Fas L,caspase3 and caspase8 in granulation tissue were detected by Western-blot and IHC.Result:1,ulcer surface healing situation Compared with the blank control group,the wound healing rate of the model control group was significantly lower,and the wound healing rate of the blank control group was significantly different from that of the model control group(P <0.01).Compared with the model group and the control group,the wound healing of Kangfuxin fluid group and MEBO group was significantly increased,with significant differences among the three groups(P <0.01),and the wound healing of MEBO group was the most significant increase.2.HE staining Compared with the blank control group,the granulation tissue cells in the model control group were disorderly discharged and the collagen fibers were superimposed in a flocculent shape.The cells of Kangfuxin fluid group were not arranged neatly.In MEBO group,only hypertrophy of fibroblasts was observed and the cells were arranged relatively neatly.Compared with MEBO group,the MEBO group was closer to the blank group.3,TUNEL test results Compared with the blank control group,the number of apoptosis in the model control group was significantly increased,and there was a significant difference between the number of apoptosis in the blank control group and the model control group(P <0.01).Compared with model control group,the number of apoptotic cells in skin granulation tissue of Kangfuxin fluid group and MEBO group was significantly decreased,and there were significant differences among the three groups(P <0.01),especially the number of apoptotic cells in MEBO group showed the lowest.4,IHC test results Fas results showed that compared with the blank control group,the model control group showed brown expression in the cytoplasm,and the expression of wound protein was significantly different between the blank control group and the model control group(P <0.01).Compared with the model control group,the expression of intracytoplasmic brown on wound surface in the Kangfuxin fluid group and MEBO group was significantly reduced,with significant differences among the three groups(P <0.01).Compared with the MEBO group,the expression of intracytoplasmic brown in the MEBO group was significantly lower.Fas L results showed that compared with the blank control group,the model control group showed brown expression in the cytoplasm,and there was significant difference in the expression of wound protein between the blank control group and the model control group(P <0.01).Compared with the model control group,the expression of intracytoplasmic brown on wound surface in the Kangfuxin fluid group and MEBO group was significantly reduced,with significant differences among the three groups(P <0.01).Compared with the MEBO group,the expression of intracytoplasmic brown in the MEBO group was significantly lower.The results of caspase3 showed that compared with the blank control group,there was brown expression in the cytoplasm of the model control group,and there was significant difference in the expression of wound protein between the blank control group and the model control group(P <0.01).Compared with the model control group,the expression of intracytoplasmic brown on wound surface in the Kangfuxin fluid group and MEBO group was significantly reduced,with significant differences among the three groups(P <0.01).Compared with the MEBO group,the expression of intracytoplasmic brown in the MEBO group was significantly lower.The results of caspase8 showed that compared with the blank control group,there was brown expression in the cytoplasm of the model control group,and there was significant difference in the expression of wound protein between the blank control group and the model control group(P <0.01).Compared with the model control group,the expression of intracytoplasmic brown on wound surface in the Kangfuxin fluid group and MEBO group was significantly reduced,with significant differences among the three groups(P <0.01).Compared with the MEBO group,the expression of intracytoplasmic brown in the MEBO group was significantly lower. 5.Results of WB test Fas results showed that the positive expression of model control group was significantly increased compared with blank control group,and there was significant difference between blank control group and model control group(P <0.01).Compared with the model control group,the positive expression of Kangfuxinye group and MEBO group was significantly decreased,and there were significant differences among the three groups(P <0.01).Compared with MEBO group,the positive expression of Kangfuxinye group and MEBO group was relatively lowest.Fas L results showed that compared with blank control group,the positive expression of model control group was significantly increased,and there was a significant difference between blank control group and model control group(P <0.01).Compared with the model control group,the positive expression of Kangfuxinye group and MEBO group was significantly decreased,and there were significant differences among the three groups(P <0.01).Compared with MEBO group,the positive expression of Kangfuxinye group and MEBO group was relatively lowest.The results of caspase3 showed that compared with the blank control group,the positive expression of model control group was significantly increased,and there was a significant difference between the blank control group and the model control group(P<0.01).Compared with the model control group,the positive expression of Kangfuxinye group and MEBO group was significantly decreased,and there were significant differences among the three groups(P <0.01).Compared with MEBO group,the positive expression of Kangfuxinye group and MEBO group was relatively lowest.The results of caspase8 showed that compared with the blank control group,the positive expression of model control group was significantly increased,and there was a significant difference between the blank control group and the model control group(P <0.01).Compared with the model control group,the positive expression of Kangfuxinye group and MEBO group was significantly decreased,and there were significant differences among the three groups(P <0.01).Compared with MEBO group,the positive expression of Kangfuxinye group and MEBO group was relatively lowest.Conclusion:1.External application of MEB can improve the healing rate of skin ulcer in diabetic mice and accelerate the healing of ulcer surface.2.MEBO can obviously inhibit the apoptosis of diabetic skin ulcer mice.3.MEB can inhibit cell apoptosis by reducing the protein expressions of Fas,Fas L,caspase3 and caspase8 in the granulation tissue of skin ulcer wound of diabetic mice. |
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