中华皮肤科杂志

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低氧对系统性硬化病患者皮肤成纤维细胞胶原蛋白合成的影响

辛崇美1, 李明2   

  1. 1. 山东临沂市皮肤病医院, 276000;
    2. 复旦大学附属中山医院皮肤科, 复旦大学上海医学院皮肤病与性病学系, 上海, 200032
  • 收稿日期:2006-07-10 出版日期:2007-06-15 发布日期:2007-06-15
  • 通讯作者: 李明,email:liming@zshospital.net E-mail:liming@zshospital.net

Effects of hypoxia on collagen synthesis in skin fibroblasts of systemic scleroderma

XIN Chong-mei1, LI Ming2   

  1. Department of Dermato-venereology, Zhongshan Hospital, Shanghai Medical College, Fudan University, Shanghai 200032, China
  • Received:2006-07-10 Online:2007-06-15 Published:2007-06-15

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摘要： 目的 研究低氧对系统性硬化病(SSc)患者和正常人皮肤成纤维细胞胶原蛋白合成及胶原基因表达的影响。方法 分别以5例SSc患者和5例正常人皮肤成纤维细胞系作为实验对象。将成纤维细胞分别放入20%正常氧和2%低氧分压环境中培养,用分光光度仪测定细胞培养基上清液中羟脯氨酸含量,并折算成胶原蛋白含量。用Trizol提取细胞总RNA,用RT-PCR方法测定其中Ⅰ型和Ⅲ型前胶原mRNA的表达。结果 培养至第6天,低氧分压条件下,SSc患者皮肤成纤维细胞培养基中胶原蛋白含量比正常氧分压条件下显著增加(t=3.97,P=0.0041);正常人皮肤成纤维细胞培养基中胶原蛋白含量此时也显著增加(t=2.63,P=0.0302)。培养至第3天,低氧分压条件下,SSc成纤维细胞Ⅰ型和Ⅲ型前胶原mRNA表达量比正常氧分压条件下均有显著增加(Ⅰ型:t=5.81,P=0.0004;Ⅲ型:t=6.44,P=0.0002);正常人皮肤成纤维细胞此时的Ⅰ型和Ⅲ型前胶原mRNA表达量与SSc患者相似,低氧分压条件下比正常氧分压条件下均有显著增加(Ⅰ型:t=4.40,P=0.0023;Ⅲ型:t=2.24,P=0.0453)。结论 低氧状态下,SSc患者和正常人皮肤成纤维细胞胶原蛋白合成均有增加,Ⅰ型和Ⅲ型前胶原mRNA表达也均增加.低氧可能是皮肤硬化的一个相关因素。

关键词: 成纤维细胞, 硬皮病, 系统性, 缺氧, 胶原

Abstract: Objective To study the effects of hypoxia on collagen synthesis and mRNA expression in systemic scleroderma(SSc)and normal dermal fibroblasts.Methods Skin specimens were resected from 5 SSc patients and 5 normal human controls.Fibroblasts were isolated from the specimens,and subjected to primary culture and passage.Then,the cells were exposed to either low(2%)or standard(20%)oxygen tension for 3 to 6 days.The collagen production level in the cultured medium was calculated based on hydroxyproline level detected by speetro-photometry.RT-PCR was used to detect the mRNA expression of proαl(Ⅰ)and proαl(Ⅲ)collagen in the fibroblasts.Results At the end of the 6th day,the concentration of collagen was significantly higher under hypoxia condition than that under normoxic condition,in the cul-tured medium of both SSc(t=3.973,P=0.0041)and normal(t=2.630,P=0.0302)skin fibroblasts. At the end of 3rd day,the mRNA expression levels of both proαl(Ⅰ)and proαl(Ⅲ)collagen were signifi-cantly higher in hypoxia condition than in normoxic condition in both SSc(typeⅠ:t=5.811,P=0.0004; typeⅢ:t=6.442,P=0.0002)and normal(typeⅠ:t=4.396,P=0.0023;typeⅢ:t=2.237,P=0.0453) skin fibroblasts.Conclusions Under hypoxia condition,the collagen synthesis,typeⅠand typeⅢprocol-lagen mRNA expression are all increased in SSc and normal skin fibroblasts.Hypoxia may be a contributory factor in skin sclerosis.

Key words: Scleroderma, systemic, Fibroblasts, Anoxia, Collagen