硫酸乙酰肝素疗法促进糖尿病大鼠创面愈合的实验研究

Experimental study of the effect of heparin sulfate on the process of wound healing in diabetic rats

目的观察硫酸乙酯肝素疗法在糖尿病大鼠创面愈合过程中对成纤维细胞生长因子(FGF)、角化细胞生长因子(KGF)、转化生长因子-β(TGF-β)和白介素-6(IL-6)水平的影响,探讨硫酸乙酰肝素促进糖尿病创面愈合的机制。方法建立糖尿病大鼠背部皮肤创面模型,采用随机表数字法将30只糖尿病大鼠分为两组:实验组(n=15)采用皮下注射0.5 ml硫酸乙酰肝素(0.3 mg/L),对照组(n=15)则采取皮下注射0.5 ml生理盐水,术后不同时期处死取材,并通过苏木精-伊红(HE)染色和免疫组化染色研究两组大鼠创面炎症反应和FGF、KGF、TGF-β和IL-6含量水平的变化。结果实验组术后5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10天中性粒细胞较前减少,伴有局部毛细血管增生;术后15天新生肉芽组织基本完成修复,而对照组创面仍有少量中性粒细胞浸润。免疫组化结果提示:术后第10和15天,实验组FGF和TGF-β阳性表达含量水平较对照组高,差异具有统计学意义(P<0.05);术后第15天,实验组KGF阳性表达含量水平较对照组高,而IL-6低于对照组,差异具有统计学意义(P<0.05)。结论硫酸乙酰肝素可能通过抑制糖尿病创面炎症反应,调节FGF、KGF、TGF-β和IL-6含量水平,促进创面肉芽组织的生长,从而达到创面愈合。

Objective To evaluate the effect heparin sulfate on fibroblast growth factor(FGF),keratinocyte growth factor(KGF), transforming growth factor-β(TGF-β), interleukin 6(IL-6) expression levels in the process of wound healing in diabetic rats. Methods Thirty diabetic rats were made full-thickness wound at the back and were randomly divided into two groups: the experimental group(n =15) accepted,0.5 ml heparin sulfate(0.3 ml/L) injection subcutaneously, and the control group accepted 0.5 ml normal saline injection subcutaneously. The hematoxylin(HE) and immunohistochemistry staining methods were applied to evaluate the histological change of wound healing at different postoperative periods. Results The HE results showed that the number of neutrophils in the experimental group was less than that in the control group with less obvious edema in the granulation tissue. The quantity of neutrophils in the experimental group at day 10 reduced and accompanied by local capillary proliferation. The basic repair with new granu-lation tissue in the experimental completed at day 15, while the wound in the control group is still accompa-nied by mild inflammatory response. The immunohistochemistry staining results showed that the positive rate of FGF and TGF-β of the diabetic wound in the experimental group was significantly higher than in the control group at 10 th and 15 th postoperative day(P<0.05);there was obvious difference in the level of KGF between the two groups at 15 th postoperative day(P<0.05); however the level of IL-6 in the experimental group was lower in the control group at the 15 th postoperative day(P<0.05). Conclusion Heparin sulfate may promote wound healing of diabetic rats by inhibiting wound inflammation and the level of FGF 、KGF、TGF-β and IL-6.