中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (11): 680-682.

• 论著 • 上一篇    下一篇

白介素1受体拮抗剂对紫外线辐射的成纤维细胞分泌基质金属蛋白酶1的影响

王小勇, 毕志刚   

  1. 南京医科大学第一附属医院皮肤科, 南京 210029
  • 收稿日期:2004-12-08 发布日期:2005-11-15
  • 通讯作者: 毕志刚, E-mail:eltonbi@2lcn.com E-mail:eltonbi@2lcn.com
  • 基金资助:
    国家自然科学基金资助项目(30271195)

Effects of the EL-1 Receptor Antagonist on Matrix Metalloproteinase 1 Expression in UV-irradiated Fibroblasts

WANG Xiao-yong, BI Zhi-gang   

  1. Department of Dermatology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2004-12-08 Published:2005-11-15

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摘要： 目的 探讨白介素1受体拮抗剂(IL—1Ra)对紫外线(UV)辐射的成纤维细胞分泌基质金属蛋白酶1(MMP-1)的影响.方法 模拟环境中UV对人体皮肤的作用方式,UVA辐射的人成纤维细胞换以UVB辐射的HaCaT培养上清继续培养.用ELISA方法检测不同处理后成纤维细胞培养上清中的MMP-1;用实时荧光定量RT-PCR方法(荧光染料掺入法)检测IL—1Ra处理后成纤维细胞C-Jun、C-Fos及内参照GAPDH的mRNA表达变化.结果 UVA(10J/cm2)辐射的成纤维细胞在加入不同剂量UVB(0、1、5、10、15mJ/cm2)辐射HaCaT的培养上清液后,其MMP—1的分泌呈增高趋势.与UVB0mJ/cm2比较,于15mJ/cm2时差异有统计学意义(t=8.413,P=0.014).而IL—1Ra以剂量依赖方式抑制成纤维细胞的MMP-1分泌.统计C—Jun和C—Fos的初始拷贝数以及其与GAPDH初始拷贝数的比值,结果显示IL—1Ra以剂量依赖方式抑制成纤维细胞C—Jun的mRNA表达,对C—Fos的mRNA表达则无显著性影响.本实验应用实时荧光定量RT—PCR,其标准曲线均有良好相关性,熔解曲线证实扩增产物是特异的.结论 UVB辐射的HaCaT培养上清液能促进UVA辐射的成纤维细胞分泌MMP-1.IL—1Ra通过抑制C-Jun的mRNA表达降低成纤维细胞的MMP—1分泌.

关键词: 间质胶原酶, 紫外线, 成纤维细胞, 白介素1受体拮抗剂

Abstract: Objective To study the effects of the IL-1 receptor antagonist (IL-1Ra) on matrix metalloproteinase 1 (MMP-1) expression in UV-irradiated fibroblasts. Methods Simulating the impact of environmental ultraviolet (UV) light on human skin, UVA-irradiated human fibroblasts were cultured in medium obtained from UVB-irradiated HaCaT cells. MMP-1 was detected by ELISA in the culture medium of fibroblasts. After treatment with IL-1Ra, the mRNA expression levels of C-Jun, C-Fos and GAPDH (internal control) of fibroblasts were measured by real-time fluorescent quantitative RT-PCR. Results Production of MMP-1 by UVA (10 J/cm2)-irradiated fibroblasts was increased in culture medium from UVB-irradiated HaCaT cells. The fibroblasts produced significantly higher levels of MMP-1 in culture medium from HaCaT cells treated without UVB than those with 15 mJ/cm2 UVB (t=8.413,P=0.014). However, IL-1Ra inhibited MMP-1 production of fibroblasts in a dose-dependent manner. Standard curves of real-time fluorescent quantitative RT-PCR showed a linear correlation between the copy number and the threshold cycle (Tc). Melting curves confirmed the specificity of PCR products. The original copy numbers of C-Jun and C-Fos as well as the ratios of the numbers to the GAPDH copy number showed that IL-1Ra inhibited the C-Jun mRNA expression of fibroblasts in a dose-dependent manner but had no significant effects on C-Fos mRNA expression. Conclusions The culture medium from UVB-irradiated HaCaT cells can promote MMP-1 production by UVA-irradiated fibroblasts. IL-1Ra reduces MMP-1 production via inhibition of C-Jun mRNA expression.

Key words: Interstitial collagenase, Ultraviolet rays, Fibroblasts, IL-1 receptor antagonist