CAJ | 학술논문

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.
목적 탐토SASPase재피부형홍반랑창피손중적표체수평급기재발병궤제중적의의.방법 무혈청배양원대각질형성세포,장제취적단백양품채용고상pH제도쌍향응효전영진행분리,응용ImageMaster 2D Platinum 5.0연건대도상진행필배분석,선취차이표체단백질점,경기질보조격광해흡부비행시간질보진행질보감정.병통과면역인적험증표체수평.결과 성공배양각질형성세포,획득중복성교호적쌍향전영도보,필배점수재1200개좌우,필배솔＞80％.감정SASPase재CLE피손각질형성세포표체승고,면역인적험증여쌍향전영결과일치.결론 피부형홍반랑창피손적발생발전가능여SASPase적이상격활화과도표체유관.
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.