Molecular biological analysis for regenerated epithelial tissues on cutaneous wound healing process

再生上皮组织对皮肤伤口愈合过程的分子生物学分析

• 批准号: 13672130
• 负责人: TAKAI Yoshiaki
• 金额: $ 2.24万
• 依托单位: Asahi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672130/
• 关键词: cutaneous wound healing; lipopolysaccharide; keratinocyte; endotoxemia; cytokine; 遺伝子発現; RT-PCR

The skin function as a non-specific general defence line against infections and injury from the environment, and is an important region as an immune defense system composed of host cells such as keratinocytes, epidernal Langerhans cells, and T lymphocytes. In the present study, we examined various gene expressions in regenerated epithelial tissues in cutaneous wound healing model mice, and the response of human keratinocytes to v IL-6, IFN-y, and keratin 6 Mrna expressions in the regenerated epithelial tissues of mice were increased just after construction of cutaneous would and their expressions continued throughout the period of observation (21 days). Cutaneous would healing in mice administered with Escherichia coli LPS was delayed as compared with that of the control mice without LPS. Human keratinocyte cell line HaCaT expressed predominantly Mrna of TNFR and IFNGR, whereas Mrna of CD14, TLR2 and TLR4 were not detected. TNF-α upregulated IL-8 and MCP-1 Mrna expression in HaCaT cells, and IFN-y downregulated IL-8 Mrna expression and upregulated MCP-1 Mrna expression. On the other hand, these cytokine-related Mrna expressions were not seen in HaCaT cells after stimulation with E. coil LPS and Staphylococcus aureus peptidoglycan. IL-8 production in culture supematants of HaCaT cells stimulation with these test specimens was coincided with their IL-8 Mrna expression. HaCaT cell growth was delayed with treatment of TNF-α and IFN-y, but not of bacterial components. Thus HaCaT cells responded to endogenous factors such as TNF-α, and IFN-y, but not exogenous factors such as E. coli LPS and S. aureus peptidoglycan. These results suggest that human keratinocytes did not directly respond to bacterial cell components, however, the cells were activated by endogenous factors induced by host immune cells stimulated with pathogenic factors

皮肤的功能是抵抗环境感染和损伤的非特异性一般防线，是由角质形成细胞、表皮朗格汉斯细胞和T淋巴细胞等宿主细胞组成的免疫防御系统的重要区域。在本研究中，我们检测了皮肤创伤愈合模型小鼠皮肤再生上皮组织中各种基因的表达，在小鼠皮肤Will构建后，人角质形成细胞对v IL-6、干扰素-γ和角蛋白6mRNA表达的反应性增加，并在整个观察期间(21天)持续表达。与未注射内毒素的对照组相比，注射内毒素的小鼠皮肤愈合延迟。人角质形成细胞株HaCaT主要表达TNFR和IFNGR的mRNA，未检测到CD14、TLR2和TLR4的表达。肿瘤坏死因子α可上调HaCaT细胞IL-8和MCP-1mRNA的表达，而干扰素-γ可下调IL-8mRNA的表达，上调MCP-1mRNA的表达。另一方面，经细菌脂多糖和金黄色葡萄球菌肽聚糖刺激后，HaCaT细胞中未见上述细胞因子相关基因的表达。这些实验标本刺激HaCaT细胞培养上清液中IL-8的产生与其IL-8mRNA的表达相一致。肿瘤坏死因子-α和干扰素-γ对HaCaT细胞生长有延缓作用，但对细菌成分无明显影响。因此，HaCaT细胞对内源性因子如肿瘤坏死因子-α和干扰素-γ有反应，而对外源因子如大肠杆菌脂多糖和金黄色葡萄球菌肽多聚糖无反应。这些结果表明，人角质形成细胞对细菌细胞成分没有直接的反应，但细胞被致病因子刺激的宿主免疫细胞诱导的内源性因素激活。

项目成果

Toshihiko Umemoto: "Chemotaxis of oral Treponemes toward sera and albumin of rabbit"Microbiology and Immunology. 45・8. 571-577 (2001)

Toshihiko Umemoto：“口腔密螺旋体对兔子血清和白蛋白的趋化性”微生物学和免疫学45・8（2001）。

Yasuyuki Asai: "Bacterial fimbriae and their peptides activate human gingival epithelial cells through Toll-like receptor 2"Infection and Immunity. 69・12. 7387-7395 (2001)

Yasuyuki Asai：“细菌菌毛及其肽通过 Toll 样受体 2 激活人类牙龈上皮细胞”感染与免疫 69・12 (2001)。

Hirai, K. et al.: "Two cases of pleomorphic adenomas of the upper lip"J. Gifu Dent. Soc. 29(1). 57-61 (2002)

Hirai, K. 等：“上唇多形性腺​​瘤两例”J.

Sumitomo, S. et al.: "CASE REPORT: Congenital sinus of the upperlip with idiopathic precocious puberty"Oral Diseases. 8. 308-309 (2002)

Sumitomo, S. 等人：“病例报告：先天性上唇窦伴特发性性早熟”口腔疾病。

小川 知彦: "Porphyromonas gingivalis合成リピドAに対するTLR欠損マウス由来歯肉線維芽細胞の認識機構"エンドトキシン研究. 4. 73-80 (2001)

小川智彦：“来自 TLR 缺陷小鼠的牙龈成纤维细胞对牙龈卟啉单胞菌合成脂质 A 的识别机制”内毒素研究 4. 73-80 (2001)。