Molecular Biological Analysis of Wound Healing

1采用db/db糖尿病小鼠和正常家兔，观察重组人胰岛素样生长因子-I（rhIGF-I）和重组人胰岛素样生长因子结合蛋白-1（rhIGFBP-1）对创面愈合的促进作用。在糖尿病C57 BL/KsJ db/db小鼠的背部和正常兔耳的内侧制备直径为6 mm的全层伤口。将不同浓度的rhIGF-I和/或rhIGFBP-1局部应用于db/db小鼠的开放伤口，每天一次，持续5天，并在创伤后一次应用于正常家兔的覆盖伤口。在第7天或第8天通过测量再上皮化（%）、肉芽组织面积（mm 2）和毛细血管数对伤口切片进行组织学评价。每种处理均加速伤口修复，以rhIGF-I + rhIGFBP-1、rhIGF-I、rhIGFBP-1和单独溶剂的降序排列。在db/db小鼠中，50 mg rhIGF-I和165 mg rhIGFBP-1（等摩尔比）联合使用显着刺激生长 ...更多信息 血管环形成（P<0.01）和毛细血管数（P<0.05）。(Ref 1）2观察中波紫外B（UVB）照射对体外培养的正常人角质形成细胞内皮素-1（ET-1）及其受体表达的影响。角质形成细胞在培养基中以2.1pg/天/105个细胞的水平分泌ET-1。UVB照射高达10 mJ/cm 2增加ET-1分泌3倍，并增强ET-1的mRNA表达。在角膜基质细胞中检测到ETA和ETB受体mRNA，并且它们的表达在5 mJ/cm 2 UVB照射后上调。(Ref 2）3采用动物模型评价糖和聚维酮碘（SP）在伤口上的局部应用。与未治疗的小鼠相比，用SP治疗的db/db小鼠的伤口显示出良好的修复率。与未治疗的兔子相比，SP治疗的正常兔子的伤口的所有三个参数都具有良好的修复率。(Ref 3）4使用体外模型研究IGF-1和（IGFBP-1）在伤口收缩中的参与。将人真皮成纤维细胞（1*105个细胞/ml）掺入无血清培养基中的猪I型胶原（最终0.21%）中。将12孔板中的成纤维细胞包埋的胶原凝胶从孔中漂浮出来，然后将各种试剂加入到测定培养基中。通过测量胶原凝胶的直径计算凝胶的表面积。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产生4.5%的凝胶收缩。同时，IGFBP-1没有引起任何显著的收缩。IGF-I和IGFBP-1以1 ng/ml的每种试剂的组合产生14%的凝胶收缩，当每种试剂单独测试时，在该浓度下没有观察到凝胶收缩。当将抗-IGF-I、抗-IGFBP-1或抗-IGF-I受体抗体加入到测定培养基中时，IGF-I和IGFBP-1诱导的凝胶收缩被显著抑制（p<0.001）。(Ref 4）更少

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