凝血酶肽类对创面愈合及表皮细胞增生与移行的作用研究

目的探讨新合成的凝血酶受体激活肽TP508对创面愈合及表皮细胞增生与移行的作用。方法SD大鼠18只，麻醉下在每一大鼠背部作4个直径1．5cm圆形皮肤切除创面。TP508治疗组9只大鼠，每1创面用TP5080.1μg(40μ1)治疗。对照组9只大鼠用40μl等渗盐水处理。用JAVAJandel和IMAGEPRO图形处理软件计算创面面积。人表皮细胞NHEK945增生采用MTT测定法和细胞计数仪直接计数法。细胞移行实验采用48孔Bovden细胞移行测定板进行，移行至滤膜下表面的细胞数以斑点密度表示。结果术后7d和14d，TP508处理组创面面积分别为盐水组的73.7％和45.4％(P<0.01)。凝血酶、TP508显著促进表皮细胞增生。培养液组细胞移行斑点密度为76.6±13.8，添加1μg／ml凝血酶和10μg／mlTP508，其斑点密度分别增至104.4±12.2和109.4±14.6结论凝血酶及TP508对大鼠皮肤切除创面愈合及人表皮细胞增生与移行均有显著作用，后者在一定程度上解释了TP508促进创面愈合的机理。

Effects of thrombln peptides on wound healing and proliferation and migration of normal human epidermal kerafinocyte (NHEK)

OBJECTIVE: To define the effects of thrombin peptides on wound healing and NHEK proliferation and migration. METHODS: A wound model was made with four 1.5 cm circular full thickness dermal excisions on the back of each Sprague-Dawley rat. 0.1 microgram (40 microliter) TP508 was applied to each circular excisional wound in 9 rats, the other 9 received saline only. Wound area was calculated with JAVA Jandel and IMAGE PRO software. NHEK945 proliferation was assessed by MTT assay and direct cell count with a Coulter Counter. Cell migration was determined by 48-well Boyden Chamber. Cells migrated onto the lower surface of the filter were assessed by a Chemi Imager 4000 Image Analyzer and expressed as spot density. RESULTS: Wound area in rats treated with TP508 was 73.7% and 45.4% of saline control on day 7 and 14, respectively. NHEK945 proliferation was accelerated after adding thrombin and TP508. The spot density of migrated cells was 76.7 plus minus 13.8 in medium alone. After adding 1 microgram/ml of thrombin and 10 microgram/ml of TP508, the spot density was 104.4 plus minus 12.2 and 109.4 plus minus 14.6, respectively. CONCLUSION: Results of this study suggest that both thrombin and TP508 have significant actions on wound healing and NHEK proliferation and migration, which is important in wound repair.