Molecular biological analyses of wound healing process and application of the gene therapy

伤口愈合过程的分子生物学分析及基因治疗的应用

• 批准号: 11470373
• 负责人: FUJII Tohru
• 金额: $ 9.09万
• 依托单位: NAGASAKI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470373/
• 关键词: wound healing; gene therapy; abnormal wound healing; Insulin-like growth factor-1, IGF-1; Leukemia Inhibitory Factor, LIF; immune tolerance; 局所サイトカイン

Among molecular biological analysis for wound healing, keloid represents the abnormal wound healing pattern. Keloid most abundantly expresses the insulin-like growth factor-1, IGF-1 as confirmed by degenerative receptor tyrosine kinase assay. IGF-1 involves in the invasiveness of keloid via post-receptor signal transduction. Moreover, keloid resists the ceramide-induced apoptosis. Ceramide normally induces the fibroblast apoptosis in dose-dependent and in time-dependent manner ; however, fibroblasts derived from keloid showed the G2 arrest by FACS cell sorter and confirmed by reduced cell number by WST-1 assay. When IGF-1 added, keloid fibroblasts resist the ceramide-induced apoptosis. This was positively correlated with IGF-1 receptor expression level. PI-3 inhibitor, Wortmaninn pre-treatment, was markedly abolished inhibition of apoptosis via IGF-1 signaling.On the other hand, on behalf of prolonged skin allograft survival, the effective modalities such as cytokine gene therapy were tested. In between BALB/c and B6D2F1 strain mice, Leukemia Inhibitory Factor, LIF plasmid cDNA was injected into the skin allograft donor. LIF expressions were observed in 24 hours and 21 days post-transplantation. The gp130, signal-transduction component of LIF, was expressed along with LIF.When B6D2F1 to BALB/c skin allografting, helper T type2 cytokine (IL-10) was observed, thus immune tolerance was indicated for skin allografting in LIF gene therapy.In bone metabolism, LIF was tested in the cranial bone defect model. LIF was efficiently enhanced the bone formation via osteocyte activation.

在创伤愈合的分子生物学分析中，瘢痕疙瘩代表了创伤愈合的异常模式。变性受体酪氨酸激酶检测证实瘢痕疙瘩中胰岛素样生长因子-1（IGF-1）表达最丰富。IGF-1通过受体后信号转导参与瘢痕疙瘩的侵袭。瘢痕疙瘩对神经酰胺诱导的细胞凋亡有抵抗作用。神经酰胺通常以剂量依赖性和时间依赖性方式诱导成纤维细胞凋亡;然而，来自瘢痕疙瘩的成纤维细胞通过流式细胞仪显示G2阻滞，并通过WST-1试验细胞数量减少证实。当加入IGF-1时，瘢痕疙瘩成纤维细胞抵抗神经酰胺诱导的凋亡。这与IGF-1受体表达水平呈正相关。PI-3抑制剂Wortmaninn预处理可明显消除IGF-1信号通路对细胞凋亡的抑制作用。另一方面，为了延长移植皮肤的存活时间，本研究还尝试了细胞因子基因治疗等有效方式。在BALB/c和B6 D2 F1系小鼠之间，将白血病抑制因子（LIF）质粒cDNA注射到皮肤移植供体中。在移植后24小时和21天观察LIF的表达。LIF的信号转导成分gp 130与LIF一起沿着表达，B6 D2 F1与BALB/c皮肤移植后可产生IL-10，提示LIF基因治疗皮肤移植可产生免疫耐受。LIF通过激活骨细胞有效地促进骨形成。

项目成果

Ishihara.H., Hirano A., Fujii T., et al.: "Keloid fibroblasts resist ceramide-induced apoptosis by overexpression of insulin-like growth factor I receptor."J Invest Dermatol. 115. 1065-1071 (2000)

Ishihara.H.、Hirano A.、Fujii T. 等人：“瘢痕疙瘩成纤维细胞通过胰岛素样生长因子 I 受体的过度表达来抵抗神经酰胺诱导的细胞凋亡。”J Invest Dermatol。

Rashid Mohammad Abdur,Sadanori Akita,Tohru Fujii,et al.: "Coadministration of Easic Fibroblast Growth Factor and Sucrose Octasulfate (Sucralfate) Facilitates the Rat Dorsal Flap Survival and Viability"Plast Reconstr Surg.. 103. 941-948 (1999)

Rashid Mohammad Abdur、Sadanori Akita、Tohru Fujii 等人：“Easic 成纤维细胞生长因子和蔗糖八硫酸盐 (Sucralfate) 的共同给药促进大鼠背皮瓣的存活和活力”Plast Reconstr Surg.. 103. 941-948 (1999)

Fujioka Masaki,Fujii Tohru,Hirano Akiyoshi: "Complete breakage of'3-D miniplates: unusual complication of osteosynthesis after sagittal split osteotomies"Scand J Plast Recons. (in press).

Fujioka Masaki、Fujii Tohru、Hirano Akiyoshi：“3-D 微型钢板完全断裂：矢状劈开截骨术后接骨术的罕见并发症”Scand J Plast Recons。

Akita, S., Fujii T.: "Necrotizing fasciitis after underlying illness and steroid intake."Ann Plas Surg. 44. 112-113 (2000)

Akita, S., Fujii T.：“潜在疾病和类固醇摄入后坏死性筋膜炎。”Ann Plas Surg。

藤井徹: "標準形成外科学"医学書院. 327 (2000)

藤井彻：“标准整形外科” Igaku Shoin 327 (2000)。