皮肤型红斑狼疮皮损中SASPase的表达和意义  

Expression and significance of skin aspartic protease in lesions of cutaneous lupus erythematosus

作　　者：方圣[1] 陈爱军[1] 单葵[1] 周汛[1] 李惠[1] 

机构地区：[1]重庆医科大学附属第一医院皮肤科,400016

出　　处：《中华皮肤科杂志》2012年第1期19-21,共3页Chinese Journal of Dermatology

摘　　要：目的 探讨SASPase在皮肤型红斑狼疮皮损中的表达水平及其在发病机制中的意义.方法 无血清培养原代角质形成细胞,将提取的蛋白样品采用固相pH梯度双向凝胶电泳进行分离,应用ImageMaster 2D Platinum 5.0软件对图像进行匹配分析,选取差异表达蛋白质点,经基质辅助激光解吸附飞行时间质谱进行质谱鉴定.并通过免疫印迹验证表达水平.结果 成功培养角质形成细胞,获得重复性较好的双向电泳图谱,匹配点数在1200个左右,匹配率＞80％.鉴定SASPase在CLE皮损角质形成细胞表达升高,免疫印迹验证与双向电泳结果一致.结论 皮肤型红斑狼疮皮损的发生发展可能与SASPase的异常激活和过度表达有关.Objective To study the expression of skin aspartic protease （SASPase） in lesions of cutaneous lupus erythematosus （CLE） and its role in the pathogenesis of CLE.Methods Skin samples were resected from the lesions and normal skin of 9 patients with CLE,including 3 cases of subacute cutaneous lupus erythematosus （SCLE）,4 cases of discoid lupus erythematosus （DLE） and 2 cases of acute cutaneous lupus erythematosus （ACLE）.Keratinocytes were isolated from the tissue samples and cultured in serum-free medium.Total proteins were extracted from the keratinocytes and separated by two-dimensional gel electrophoresis.ImageMaster 2D analysis software was used to assess differentially expressed proteins in keratinocytes between the lesional and normal skin,which were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）.The expression levels of SASPase were further determined by Western blot.The data were analyzed statistically by Student＇s t test.Results Keratinocytes were isolated from the tissue samples and successfully cultured in vitro.Two-dimensional electrophoresis profiles of proteins from the keratinocytes were obtained with high resolution and reproducibility,and the average matching protein spots were about 1200 with the matching rate higher than 80%.As Western blot showed,the relative expression level of SASPase was 0.463 ± 0.018 in keratinocytes from the lesional skin,and 0.145 ± 0.011 in those from the normal skin （P 〈 0.05）.The Western blot results were consistent with those of two-dimensional electrophoresis.Conclusion The initiation and progression of CLE seem to be associated with the abnormal activation and overexpression of SASPase.

关 键 词：红斑狼疮 皮肤 SASPase 角质形成细胞 

分 类 号：R593.241[医药卫生—内科学]