Molecular Biological Analysis of Wound Healing

1用糖尿病小鼠和正常兔评价重组人胰岛素样生长因子-I和重组人胰岛素样生长因子结合蛋白-1对创面愈合的促进作用。在C57BL/KSJ db/db糖尿病小鼠背部和正常兔耳内侧制备直径6 mm的全层创面。将不同浓度的重组人胰岛素样生长因子-I和/或重组人胰岛素样生长因子结合蛋白-1局部应用于db/db小鼠开放创面，每日1次，连续5天，正常兔创面伤后局部应用1次。术后第7天或第8天取材，通过测量再上皮化率(%)、肉芽组织面积(Mm2)和毛细血管数进行组织学评价。RhIGF-I+rhIGFBP-1组、rhIGF-I组、rhIGFBP-1组、单独用药组均能促进创面修复。在db/db小鼠中，50毫克重组人胰岛素样生长因子-I和165毫克重组人胰岛素样生长因子结合蛋白-1(等摩尔比)显著刺激GR…。更多的绒毛组织形成(p&lt；0.01)和毛细血管数量(p&lt；0.05)。[参考文献1]2用体外培养的正常人角质形成细胞观察中波紫外线(UVB)照射对人角质形成细胞内皮素-1(ET-1)及其受体表达的影响。角质形成细胞分泌ET-1的量为2.1pg/天/105个细胞。UVB照射至10mJ/cm2时，ET-1的分泌增加3倍，ET-1的mRNA表达增强。ETA和ETB受体mRNAs在角质形成细胞中均有表达，5mJ/cm2 UVB照射可上调其表达。(参考文献2)3用动物模型评估糖和聚维酮碘(SP)在创面上的局部应用。与未处理的小鼠相比，用SP处理的db/db小鼠的伤口显示出良好的修复率。与未处理兔相比，SP处理的正常兔创面在三个参数上均有良好的修复率。(参考文献3)4利用体外模型研究IGF-I和IGFBP-1在伤口收缩中的作用。人真皮成纤维细胞(1×105个/ml)在无血清条件下掺入猪I型胶原(最终浓度为0.21%)。将包埋在12孔板中的成纤维细胞胶原凝胶从孔中漂浮出来，然后将各种试剂加入到测定介质中。通过测量胶原凝胶的直径来计算凝胶的表面积。高剂量(30和100 ng/ml)的IGF-I分别显示6.8%和7.7%的凝胶收缩，10 ng/ml的DES(1-3)IGF-I则产生4.5%的凝胶收缩。同时，IGFBP-1没有引起任何明显的收缩。IGF-I和IGFBP-1的组合在每个试剂的浓度为1 ng/ml时，当单独测试每个试剂时没有观察到凝胶收缩，产生14%的凝胶收缩。当加入抗IGFBP-1、抗IGFBP-1或抗IGFBP-I受体抗体时，IGF-I和IGFBP-1诱导的凝胶收缩明显受到抑制(p&lt；0.001)。(参考文献4)少

1 The stimulatory effect of recombinant human insulin-like growth factor-I (rhIGF-I) and recombinant human insulin-like growth factor-binding protein-1 (rhIGFBP-1) on wound healing was assessed using diabetic db/db mice and normal rabbits. Full thickness wounds of 6 mm diameter were prepared on the backs of diabetic C57BL/KsJ db/db mice and on the inner sides of normal rabbit ears. Various concentrations of rhIGF-I and/or rhIGFBP-1 were applied locally to the open wounds of db/db mice once daily for 5 days and to the covered wounds of normal rabbits once after wounding. Sections of the wounds were evaluated histologically on the 7th or 8th day by measuring reepithelialization (%), area of granulation tissue (mm2) and capillary numbers. Wound repair was accelerated by each of the treatments in descending order of rhIGF-I plus rhIGFBP-1, rhIGF-I,rhIGFBP-1, and vehicle alone. In db/db mice, the combination of 50 mg rhIGF-I and 165 mg rhIGFBP-1 (equimolar ratio) significantly stimulated gr … More anulation tissue formation (p<0.01) and capillary numbers (p<0.05). (Ref 1)2 The effect of ultraviolet B (UVB) irradiation on Endothelin-1 (ET-1) and ET receptor expressions was examined using cultured normal human keratinocytes. Keratinocytes secreted ET-1 in the medium at a level of 2.1 pg/day/105 cells. UVB irradiation up to 10 mJ/cm2 increased ET-1 secretion 3-fold, and potenitated expressions of mRNA for ET-1. Both ETA and ETB receptor mRNAs were detected in kerationocytes, and their expressions were up-regulated by 5 mJ/cm2 UVB irradiation. (Ref 2)3 Topical application of sugar and povidone iodine (SP) on wounds was assessed using animal models. The wounds in db/db mice treated with SP displayd good repair rates compared to those for non-treated mice. The wounds in normal rabbits treated with SP had good repair rates for all three parameters compared with those for non-treated rabbits. (Ref 3)4 Involvement of IGF-I and (IGFBP-1 in wound contraction was studied using an in vitro model. Human dermal fibroblasts (1*105 cells/ml) were incorporated into a porcine type I collagen (0.21% final) in serum-free medium. The fibroblast-embedded collagen gels in a 12-well plate were floated from the well, and various reagents were then added to the assay medium. The surface area of the gel was calculated by measuring the diameters of the collagen gel. IGF-I at high doses (30 and 100 ng/ml) revealed 6.8% and 7.7% gel contraction, respectively, and des (1-3) IGF-I at 10 ng/ml produced a 4.5% gel contraction. Meanwhile, IGFBP-1 did not induce any significant contraction. A combination of IGF-I and IGFBP-1 at 1 ng/ml of each reagent, a concentration at which gel contraction was not observed when each of the reagents was tested individually, produced a 14% gel contraction. When anti-IGF-I,anti-IGFBP-1 or anti-IGF-I receptor antibody was added to the assay medium, the IGF-I and IGFBP-1-induced gel contraction was significantly suppressed (p<0.001). (Ref 4) Less