| Objectives:To evaluate the effect of artesunate(ART)on wound healing and detect the expression of matrix metalloproteinase2(MMP-2),matrix metalloproteinase.9(MMP-9),transforming growth factor beta 1(TGF-?1),epidermal growth factor(EGF)in diabetic rat model.It may provide a possible way topromoteskin wound healing in diabetes.Methods:50 healthy male SD rats of 180-200 g were selected and fed for one week.Ten of rats were randomly selected into healthy control group and forty rats were randomly selected to establish diabetic model which were intraperitoneally injected of streptozotocin(STZ).The above rats were divided into 5 groups:Normal rats saline group,diabetic rats saline group,diabetic rats 0.5%ART group,diabetic rats 1.0%ART group,diabetic rats rhEGF group.Fasting blood glucose and body weight were monitored.After establishing diabetes model,two round full-thickness skin wounds with a diameter of 1 cm were prepared on the back in rats,and the drug was used from the next day.Saline was spread on the wounds of the rats in normal ratssaline group and diabetic rats saline group.0.5%ART and 1.0%ART were spread on the wounds of the rats in diabetic rats 0.5%ART group,diabetic rats 1.0%ART group.RhEGF was spread on the wounds of the rats in diabetic rats rhEGF group.The wounds were photographed on the 1st,3rd,5th,7th,10th and 14th day after smearing drugs,and the unhealed area of the wounds were measured by ImageJ.According to the wound healing rate=(original wound area-unhealed wound area)/original Wound area × 100%,we could calculate the wound healing rate.Rats were sacrificed after 15 days and skin wounds tissues were collected.The pathophysiological morphology changes of skin were observed by hematoxylin-eosin staining(HE).Collagen changes were observed by Masson collagen staining.The value of MVD and the expressions of MMP-2,MMP-9,TGF-?1 and EGF in skin wounds were detected by immunohistochemistry(IHC).The mRNA expression of MMP-2,MMP-9,TGF-?1 and EGF were detected by fluorescence quantitative RT-PCR.Results:1.One week and two weeks after injecting STZ,the fasting blood glucose of diabetic rats in each group increased significantly,which was higher than 16.7 mmol/L.The body weights of diabetic rats in each group were significantly lower than that of normal rats.There was not significant difference in fasting blood glucose and body weights among diabetic groups.One week and two weeks after making wounds,the fasting blood glucose of diabetic rats in each group was still higher than that of normal rats,and their body weights decreased significantly.There was not significant difference in the fasting blood glucose and body weights among diabetic groups.2.On the 3rd day,there were not significant difference in wound healing rate among normal saline group,diabetic saline group,diabetic 0.5%ART group,diabetic 1.0%ART group and diabetic rhEGF group.On the 5th day,there were not significant difference in wound healing rate among normal saline group,diabetic 1.0%ART group,diabetic rhEGF group and diabetic saline group,while wounds in diabetic 0.5%ART group had higher wound healing rate than that in diabetic saline group.On the 7th day,there were not significant difference in wound healing rate among normal saline group,diabetic 1.0%ART group and diabetic saline group,while wounds in diabetic 0.5%ART group and diabetic rhEGF group had higher healing rate than that in diabetic saline group.On the 10th day,compared with diabetic saline group,the wound healing rate in normal saline group and diabetic saline group increased significantly,and the wound healing rate in diabetic 0.5%ART group was higher than that in diabetic 1.0%ART group.After 14 days of treatment,the skin wounds healed well in normal saline group,and the areas of wounds area decreased significantly.Wound healing rate in normal saline group was significantly higher than that in diabetic saline group.The areas of wounds in diabetic saline group was larger,while the wound healing rate was higher in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group than that in diabetic saline group.3.The observation of wounds by HE staining showed that the epidermal migration in diabetic saline group was imcomplete with less fibroblasts and neovascularization,more inflammatory cells and loose collagen fibers.The wounds in normal saline group healed well,fibroblasts were more active,and had more granulation tissues and neovascularization and less inflammatory cells.The wounds in diabetic 0.5%ART group,diabetic 1.0%ART group healed better than that in diabetic saline group and had more granulation tissues and less inflammatory cells.The wounds in diabetic rhEGF group healed well and had more neovascularization and more active fibroblasts.The observation of wounds bymasson collagen staining showed that the blue collagen fibers in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group were more dense and orderly than that in diabetic saline group.4.The observation of wounds by immunohistochemistry showed that the expression of MMP-2 and MMP-9 in skin wounds in diabetic saline group were higher than that in normal saline group.Compared with diabetic saline group,the expression of MMP-2 and MMP-9 in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group were significantly lower.The value of MVD and the expression of TGF-?1 and EGF in diabetic saline group were lower than that in normal saline group.Compared with diabetic saline group,the expression of TGF-?1 and EGF in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group were significantly higher.Besides,compared with diabetic saline group,the value of MVD in diabetic rhEGF group was significantly higher.5.The results of fluorescence quantitative RT-PCR showed that the mRNA expression of MMP-2 and MMP-9 in diabetic saline group were higher than that in normal saline group,and the mRNA expression of MMP-2 and MMP-9 in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group were lower than that in diabetic saline group,the mRNA expression of TGF-?1 and EGF in diabetic saline group were lower than that in normal saline group,the mRNA expression of TGF-?1 and EGF in diabetic 0.5%ART group,diabetic 1.0%ART group,diabetic rhEGF group were higher than that in diabetic saline group.Conclusion:1.Compared with the wounds in normal rats,wounds in diabetic rats healed well,and the expression of MMP-2,MMP-9 were higher while the expression of TGF-?1 and EGF were lower in wounds.2.ART and rhEGF cound promote the wound healing in diabetic rats and it may have effects by down-regulating the expression of MMP-2,MMP-9 and up-regulating the expression of TGF-?1 and EGF. |
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