1,25-二羟基维生素D(3) 保护人类角质形成细胞免受紫外线B诱导的损伤：细胞活力/增殖、DNA损伤及修复的体外分析
1,25-dihydroxyvitamin D(3) protects human keratinocytes against UV-B-induced damage: In vitro analysis of cell viability/proliferation, DNA-damage and -repair.
作者信息
Trémezaygues Lea, Seifert Markus, Tilgen Wolfgang, Reichrath Jörg
机构信息
Department of Dermatology; The Saarland University Hospital; Homburg/Saar, Germany.
出版信息
Dermatoendocrinol. 2009 Jul;1(4):239-45. doi: 10.4161/derm.1.4.9705.
The skin is the only organ that has the capacity to photo-synthesize the biological active vitamin D metabolite 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] from 7-dehydocholesterol (7-DHC), following exposure to ultraviolet (UV)-B irradiation. The aim of the present work was to investigate the capacity of 1,25(OH)(2)D(3) to protect human keratinocytes (HaCaT) and squamous cell carcinoma cell lines (SCL-1) against the hazardous effects of UV-B irradiation. Human keratinocytes (HaCaT) and squamous cell carcinoma cell lines (SCL-1) were pretreated with 1,25(OH)(2)D(3) over 48 hours and then irradiated once with UVB-radiation. We evaluated the results of several assays (colony-forming-unit-culture assay, WST-1-assay and crystal violet assay), comparing viability/proliferation in 1,25(OH)(2)D(3)-pretreated cells with controls that were pretreated with the carrier substance ethanol alone. Additionally, we analyzed the effects of 1,25(OH)(2)D(3) on UV-induced DNA damage in HaCaT-keratinocytes by detection of cyclobutane pyrimidine dimers (CPDs) via dot blot analysis. We prove that 1,25(OH)(2)D(3), in a concentration of 10(-7) M, protects human keratinocytes (HaCaT) as well as squamous cell carcinoma cell lines (SCL-1) against the hazardous effects of UV-B-radiation (100 J/cm(2)-1,000 J/cm(2)) in vitro. Moreover, we demonstrate that the number of CPDs induced in HaCaT-keratinocytes after irradiation with UV-B (100 J/cm(2)-1,000 J/cm(2)) was decreased after pretreatment with 1,25(OH)(2)D(3), as compared to carrier-treated controls. Analysis of the time course revealed that the elimination of UV-B-induced DNA-damage in HaCaT-keratinocytes occurs quicker when cells are pretreated with 1,25(OH)(2)D(3) (as compared to controls). To put it in a nutshell, our data support the hypothesis that 1,25(OH)(2)D(3) protects cultured human keratinocytes against the hazardous effects of UV-B radiation.
皮肤是唯一能够在暴露于紫外线B（UV-B）照射后，将生物活性维生素D代谢物1,25-二羟基维生素D(3) [1,25(OH)(2)D(3)] 从7-脱氢胆固醇（7-DHC）光合成的器官。本研究的目的是调查1,25(OH)(2)D(3) 保护人角质形成细胞（HaCaT）和鳞状细胞癌细胞系（SCL-1）免受UV-B照射有害影响的能力。人角质形成细胞（HaCaT）和鳞状细胞癌细胞系（SCL-1）用1,25(OH)(2)D(3) 预处理48小时，然后用UVB辐射照射一次。我们评估了几种检测方法（集落形成单位培养检测、WST-1检测和结晶紫检测）的结果，将1,25(OH)(2)D(3) 预处理细胞的活力/增殖与仅用载体物质乙醇预处理的对照进行比较。此外，我们通过斑点印迹分析检测环丁烷嘧啶二聚体（CPD），分析了1,25(OH)(2)D(3) 对HaCaT角质形成细胞中UV诱导的DNA损伤的影响。我们证明，浓度为10(-7) M的1,25(OH)(2)D(3) 在体外可保护人角质形成细胞（HaCaT）以及鳞状细胞癌细胞系（SCL-1）免受UV-B辐射（100 J/cm(2)-1,000 J/cm(2)）的有害影响。此外，我们证明，与载体处理的对照相比，用1,25(OH)(2)D(3) 预处理后，HaCaT角质形成细胞在UV-B（100 J/cm(2)-1,000 J/cm(2)）照射后诱导的CPD数量减少。时间进程分析表明，当细胞用1,25(OH)(2)D(3) 预处理时，HaCaT角质形成细胞中UV-B诱导的DNA损伤的消除比对照更快。简而言之，我们的数据支持1,25(OH)(2)D(3) 保护培养的人角质形成细胞免受UV-B辐射有害影响这一假设。
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