重组胸腺素β4调节ICAM-1、MMP-2和LN-5促进创伤愈合的实验研究  

An Experimental Study: Recombinant Thymosin β4 Accelerates Skin Wound Healing by Regulating ICAM-1, MMP-2 and LN-5

作　　者：李艳[1] 王冠[1] 于虎[1] 杨小凡[1] 许松山[2] 聂李亚[2] 车永哲[1] 

机构地区：[1]南开大学医学院,天津市300071 [2]北京诺思兰德生物技术有限公司,北京市100085

出　　处：《组织工程与重建外科杂志》2008年第3期142-145,163,共5页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家大学生创新性实验计划(NK0707)

摘　　要：目的探讨重组胸腺素β4(Thymosin β4,Tβ4)通过改善创面基质环境促进大鼠皮肤创伤愈合的作用机制。方法在大鼠背部制作直径8mm的全层皮肤损伤模型,动物分为对照组(PBS)、Tβ4低剂量组、Tβ4中剂量组、Tβ4高剂量组,每组5只,每天两次给药,2d、4d、7d取材,行ICAM-1(Intercellular adhesion molecule-1)、MMP-2(Matrix metallopoproteinase-2)、LN-5(Laminin-5)的免疫组化并进行平均积分光密度(IOD)分析。结果经Tβ4处理后的创面,ICAM-1阳性表达多,早期中、高剂量组表达较强(p<0.01),中期均减弱,高剂量组相对较强;晚期各组表达增强,高剂量组最强(p<0.01)。MMP-2早期对照组表达最高(p<0.01),高剂量组最弱(p<0.01);中期中剂量组表达明显(p<0.05);晚期基质阳性明显增多,对照组表达最强(p<0.01),高剂量组最弱(p<0.01)。LN-5早期各组均可见较多的阳性表达,对照组与低剂量阳性强(p<0.01),中剂量组阳性最少;中期中剂量组表达最强(p<0.01);晚期中剂量组仍然有较强的阳性表达(p<0.01)。结论Tβ4可能是通过调节ICAM-1、MMP-2、LN-5的表达,改善创面基质环境,促进皮肤创伤愈合。Objective To study the mechanisms of recombinant thymosin β4 （Tβ4） accelerates skin wound healing in rats by regulating expression of ICAM-1,MMP-2 and LN-5, which are the key cytokines of ECM reforming. Methods Two full thickness 8 mm punch wounds were made on the dorsal surface each side of the rat. Tβ4 （Beijing Northland Biotech, China） was applied topically at the time of wounding （2 μg, 6 μg, 18μg in 50 μl PBS） and every 12h post-wounding. Control group received identical amounts of phosphate buffered saline （PBS） at the same time. At days 2, 4 and 7 post-wounding, materials were collected from wound tissue and fixed in 4 % paraformaldehyde for 24 h. The paraffin sections which from the middle of the wound were performed ICAM-1, MMP-2, LN-5 immunohistochemistry stain. This experiment was repeated twice using five rats for each test condition. Results Compared with control, ICAM-1 immuno-positive cells and vessels were the weakest at 4d post-wounding, higher positive at 2 d post-wounding and the peak positive at 7 d post-wounding while an excess expression in Tβ4 18μg group （p〈0.01）. MMP-2 immuno-stain were showed maintain high level, linear intensive density and lots of positive cells and ECM expressing in Tβ4 6μg group. LN-5 immunohistochemistry stain were showed a little more positive expressing early, including positive cells and ECM, but Tβ4 6μg group were expressed fewer, only at the borderline and bottom of the wound, while a significant increasing at 4d post-wounding （p 〈0.01） and a relative high expressing with cells in good directivity at 7d post-wounding （p〈0.01）. Conclusion Tβ4 may be modulate ICAM-1, MMP-2 and LN-5 proteic expression to regulate the reformation of ECM accelerating skin wound healing.

关 键 词：重组胸腺素β4 皮肤创伤愈合 细胞间粘附分子-1 基质金属蛋白酶-2 层粘连蛋白-5 

分 类 号：R641[医药卫生—外科学]