Molecular Biological Analysis of Wound Healing

伤口愈合的分子生物学分析

• 批准号: 07670967
• 负责人: TSUBOI Ryoji
• 金额: $ 1.22万
• 依托单位: Juntendo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670967/
• 关键词: wound healing; animal model; endothelin; IGF-I; IGFBP-1; gel contraction; diabetes; povidone iodine; IGF

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

1 使用糖尿病 db/db 小鼠和正常兔子评估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) 对伤口愈合的刺激作用。在糖尿病C57BL/KsJ db/db小鼠的背部和正常兔耳的内侧制备直径6mm的全层伤口。将不同浓度的rhIGF-I和/或rhIGFBP-1局部施用到db/db小鼠的开放性伤口上，每天一次，持续5天，并且在受伤后将不同浓度的rhIGF-I和/或rhIGFBP-1局部施用到正常兔的覆盖伤口上一次。在第7天或第8天通过测量再上皮化(%)、肉芽组织面积(mm2)和毛细血管数量对伤口切片进行组织学评估。按照降序排列的rhIGF-I加rhIGFBP-1、rhIGF-I、rhIGFBP-1和单独载体的每种治疗加速了伤口修复。在 db/db 小鼠中，50 mg rhIGF-I 和 165 mg rhIGFBP-1（等摩尔比）的组合显着刺激血管组织形成 (p<0.01) 和毛细血管数量 (p<0.05)。 (参考文献1)2 使用培养的正常人角质形成细胞检查紫外线B (UVB) 照射对内皮素-1 (ET-1) 和ET 受体表达的影响。角质形成细胞在培养基中以2.1皮克/天/105个细胞的水平分泌ET-1。高达 10 mJ/cm2 的 UVB 照射使 ET-1 分泌增加 3 倍，并增强 ET-1 mRNA 的表达。在角质形成细胞中检测到ETA和ETB受体mRNA，并且它们的表达在5 mJ/cm2 UVB照射下上调。 （参考文献 2）3 使用动物模型评估了糖和聚维酮碘 (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，用 SP 治疗的正常兔子的伤口在所有三个参数上都有良好的修复率。 (参考文献 3)4 使用体外模型研究 IGF-I 和 (IGFBP-1) 参与伤口收缩的情况。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 型胶原蛋白 (终浓度 0.21%) 中。将 12 孔板中的成纤维细胞包埋的胶原蛋白凝胶从孔中漂浮出来，并加入各种试剂 然后添加到测定介质中。通过测量胶原凝胶的直径来计算凝胶的表面积。高剂量（30 ng/ml 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没有引起任何明显的收缩。 每种试剂1ng/ml的IGF-I和IGFBP-1的组合（当单独测试每种试剂时未观察到凝胶收缩的浓度）产生14%的凝胶收缩。当抗 IGF-I、抗 IGFBP-1 或抗 IGF-I 受体抗体添加到测定介质中时，IGF-I 和 IGFBP-1 诱导的凝胶 收缩被显着抑制（p<0.001）。 （参考文献 4）更少

项目成果

R.Tsuboi: "Ultraviolet B irradiation increases endothelin-1 and endothelin receptor expression in cultured human keratinocytes." FEBS Lett. 371. 188-190 (1995)

R.Tsuboi：“紫外线 B 照射会增加培养的人角质形成细胞中内皮素 1 和内皮素受体的表达。”

石重 明: "糖尿病マウスに作製した皮膚潰瘍に対する精製白糖ポビドンヨードの創傷治癒促進効果:特に病理組織学的検討を中心に" 日皮会誌. 106. 403-408 (1996)

Akira Ishishige：“纯化白糖聚维酮碘对糖尿病小鼠皮肤溃疡的伤口愈合促进作用：关注组织病理学检查”日本皮肤病学会杂志 106. 403-408 (1996)。

Tsuboi R,Shi CM,Sato C,Cox CN,Ogawa H: "Co-administration of insulin-like growth factor (IGF)-I and IGF-binding protein-1 stimulates wound healing in animal models." J Invest Dermatol. 104. 199-203 (1995)

Tsuboi R、Shi CM、Sato C、Cox CN、Okawa H：“胰岛素样生长因子 (IGF)-I 和 IGF 结合蛋白-1 的共同给药可刺激动物模型中的伤口愈合。”

坪井 良治: "創傷治癒の基礎と臨床." 前実績研修講習会必須Aコース.日本皮膚科学会, 27 (1995)

Ryoji Tsuboi：“伤口愈合的基础和临床实践。”日本皮肤病学会的先决条件培训课程，27（1995）。

Lee Y-R,Oshita Y,Tsuboi R,Ogawa H: "Combination of insulin-like growth factor (IGF)-I and IGF-binding protein-1 promotes fibroblast-embedded collagen gel contraction." Endocrinology. 137. 1-6 (1996)

Lee Y-R、Oshita Y、Tsuboi R、Okawa H：“胰岛素样生长因子 (IGF)-I 和 IGF 结合蛋白-1 的组合可促进成纤维细胞嵌入的胶原凝胶收缩。”