| Objective: This experiment through the use of low-intensity ultrasonic to irradiate the wound of diabetic rats, then observe the process of wound healing and changes of cytokines. Base on the wound healing, cell proliferation, angiogenesis and cell signal transduction to explore the feasibility of low-intensity ultrasound in the treatment of diabetic refractory wound. To provide a scientific and effective basis for the new method in the treatment of diabetic refractory wound.Methods: 60 healthy adult male Wistar rats were randomly divided into four groups.Group A: healthy control group; Group B: blank control group of diabetic rats; C: Low-intensity ultrasound treatment group of diabetic rats; Group D: routine treatment group of diabetic rats. Use streptozotocin(STZ) to induce diabetic rat model and then construct the diabetic rat model of full thickness skin defect. In the first day after treatment and 7 days after treatment, 14 days and 21 days recording the process of wound healing.Cuting the wound tissue to observe the granulation tissue growth and fibroblast proliferation status in HE staining; Immunohistochemistry to detect the local expression of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in wound tissues in different time; Real-time PCR to detect the expression of TGF-β1mRNA and Smad3 mRNA in wound tissue. The data were analyzed by SPSS 17 software.Results:1.General situation: after diabetic rat model was constructed successfully, there appeared polydipsia, polyphagia, polyuria symptoms in diabetic rats, hair dull, body weight decreased significantly and relative reduction of subcutaneous fat; rats in normal group with sensitive reaction and normal diet and weight gain.2.HE staining showed that after the low-intensity ultrasound irradiation treatment, diabetic rats fibroblast proliferation in the wound were more obviously than other groups,fibrous tissue increased, more abundant neovascularization.3.Immunohistochemical staining showed that VEGF and bFGF began to express in the first day after trauma forming.7 days after treatment, group A was significantly higher than that of group B and group D(P < 0.05),group A is higher than that of the group C(P > 0.05); 14 days after treatment, group C expression reached a peak, was significantly higher than that in group B and group D(P < 0.05);21days after treatment, VEGF and bFGF expression in all group were declined, group C expression of VEGF and bFGF were still obviously higher than other three groups(P < 0.05).4.Real-time PCR showed that, on the first day of sampling, each relative expression was low, between groups had no significant difference(P > 0.05); 7 days after treatment, group A relative expression amount was increasing, group C relative expression was the highest, and group A had no significant difference compare with group C(P > 0.05); 14 days after treatment, each group expression of TGF-β1mRNA、smad3mRNA reached the highest and group C was significantly higher than that in other groups(P<0.05); 21 days after treatment,all groups relative expression quantity were reduced, group C was still significantly higher than that in the other groups(P < 0.05).Conclusions:1.Low-intensity ultrasound treatment of diabetic rat wound can improve the wound healing rate, promote granulation tissue growth and fibroblast proliferation.2.Low intensity ultrasound treatment of diabetic rat wound can promote local vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) expression.3.Low intensity ultrasound treatment of diabetic rat wound can promote the expression of transforming growth factor beta 1(TGF-β1) and Smad3 in local tissue.4.Low intensity ultrasound has the characteristics of no direct damage and non-invasive. |
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