硫酸乙酰肝素基质疗法对糖尿病大鼠创面愈合影响的组织学研究

摘要

目的 研究硫酸乙酰肝素对糖尿病大鼠创面愈合的影响.方法 36只雄性糖尿病大鼠随机数字法分为两组:实验组(n=18)采用皮下注射0.5ml硫酸乙酰肝素(0.3mg/ml),对照组(n=18)则皮下注射0.5ml生理盐水.两组大鼠麻醉成功后在其背部制作一直径为20 mm的圆形伤口,观察创面愈合情况,并通过苏木精-伊红和免疫组化染色研究两组大鼠伤口肉芽组织厚度和炎症反应情况.结果术后第4、6、8、10、12和14天时实验组伤口愈合率明显快于对照组(P<0.05).实验组术后3天中性粒细胞相对于对照组较少,肉芽组织无明显水肿;术后7天时中性粒细胞较前减少,伴有局部毛细血管的增生;术后14天时新生肉芽组织基本修复完成,而对照组伤口仍伴有轻度炎症反应.免疫组化结果提示术后第14天实验组伤口肿瘤坏死因子-α阳性面积较对照组明显降低(561.4±18.7 vs 685.0±31.6,P=0.004),血管内皮生长因子阳性面积较对照组明显升高(688.1±19.3 vs 584.2±14.6,P=0.002).结论硫酸乙酰肝素可能通过增加血管新生和抑制伤口炎症反应,从而促进创面的愈合.%Objective To observe the effect of heparan sulfate on wound healing in diabetic rats. Methods Thirty-six male diabetic rats were randomly divided into two groups: 0.5 ml heparan sulfate (0.3 mg/ml) was injected subcutaneously in rats of the experimental group (n=18), 0.5ml normal saline was injected subcutaneously in rats of the control group (n=18). A round wound with a diameter of 20 mm and a depth of about 3mm was produced in the epithelium tissue in the back of rats after the success of anesthesia. The thickness of granulation tissue and the inflammatory reaction were observed by hematoxylin ( HE ) and immunohistochemical staining. Results The wound healing rate in the experimental group was significantly higher than that in the control group at 4, 6, 8, 10, 12 and 14 days after operation (P<0.05). The number of neutrophils in the experimental group was less than that in the control group, and no significant edema was found in the granulation tissue. The quantity of neutrophils in the experimental group at day 7 was reduced accompanied by local capillary proliferation. The basic repair with new granulation tissue in the experimental group completed at day 14, while the wound in the control group is still accompanied by mild inflammatory response . Immunohistochemical staining showed that the positive rate of TNF-α of the wound in the experimental group was significantly lower than that in the control group at day 14 (561.4 ± 18.7 vs 685.0 ± 31.6, t=5.830, P=0.004); the positive area of VEGF of the wound in the experimental group was significantly higher than that in the control group at day 14 (688.1 ± 19.3 vs 584.2 ± 14.6, t=7.436, P=0.002).Conclusion Heparin sulfate may promote wound healing by increasing angiogenesis and inhibiting wound inflammation.