芥子气诱导小鼠耳廓皮肤差异蛋白的鉴定  

Identification of differential proteins of pinnal skin of mice poisoned by sulfur mustard

作　　者：安文华[1] 钟玉绪[1] 应翔宇[1] 时辉宁[1] 应莺[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国药理学与毒理学杂志》2006年第2期152-156,共5页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的 通过芥子气染毒后小鼠耳廓皮肤蛋白质组的改变。探索芥子气皮肤损伤的分子机制。方法 5μL（160g·L^-1）芥子气经小鼠耳廓内侧皮肤染毒，于染毒后6和24h制备蛋白样品，采用双向电泳技术分离蛋白样品，ImageMaster 5．01软件处理双向电泳图谱，基质辅助激光解吸附飞行时间质谱分析差异蛋白点的肽质量指纹谱。结果 正常对照组、染毒后6和24h组分别检测到1442，1451和1158个蛋白质点，各组蛋白质的匹配百分比为64．5％-82．0％。共鉴定了15个差异蛋白。主要与过氧化应激、细胞凋亡及能量代谢等相关。染毒后的表达上升的蛋白有载脂蛋白A-I和前白蛋白；表达下降的蛋白有：过氧化物氧还酶6、超氧化物歧化酶、羰基还原酶3、肌动蛋白、谷胱甘肽S．转移酶Mu2和△^3,5，△^2,4-二烯酰辅酶A异构酶；6h时表达上升，24h时下降的蛋白有：丙糖磷酸异构酶、脂肪酸结合蛋白、微丝结合蛋白-1、肌动蛋白解聚因子和蛋白突触结合蛋白样蛋白-3的剪接同工型4。6h表达下降，24h表达上升的蛋白是半胱氨酸天冬氨酸蛋白酶抑制剂B。钙粒蛋白B仅出现于染毒后6h组，对照组与染毒后24h组没有出现。结论 芥子气染毒能诱导小鼠皮肤蛋白表达谱的改变。AIM To investigate the molecular mechanism of skin lesion in sulfur mustard-poisoned mice by differential proteomic analysis. METHODS Mouse ears were isolated from normal mice and mice treated topically with sulfur mustard（5 μL,160 g·L^-1 ）. The total protein samples used in 2-D electrophoresis were prepared by discarding cartilage and ultracentrifugation. The protein maps were compared with ImageMaster 5.01 analysis software and analyzed with matrix-assisted laser desorption ionization-time of flight mass spectrometry （MALDI-TOF-MS）. RESULTS The number of protein spots in normal, 6-h post-poisoned and 24-h post-poisoned groups were 1442,1451 and 1158, respectively. The matching percentage of protein spots between different groups were 64.5% - 82.0%. Fifteen different proteins identified by peptide mass fingerprinting were involved in cellular events, such as oxidative stress responses, apoptosis and energy metabolism. Among.them, the expression of apolipoprotein A- I and prealbumin was upregulated in post-poisoned group, and the expression peroxiredoxin 6, superoxide dismutase [ Cu- Zn], carbonyl reductase 3, actin, glutathione S-transferase Mu2, △^3,5 and △^2,4-dienoyl-CoA isomerase was decreased in post-poisoned group. Compared with the normal group, triosephosphate isomerase, profilin-1, actin-depolymerizing factor （Destrin） , splice isoform 4 of synaptotagmin-like protein 3 and cystatin B （stefin B） were increased in 6-h post-poisoned group and decreased in 24-h post-poisoned group. Fatty acid-binding protein was decreased in 6-h post-poisoned group and increased in 24-h post-poisoned group. Calgranulin B was found in 6-h post-poisoned group, but not in normal group and 24-h post-poisoned group. CONCLUSION Sulfur mustard can alter the protein expression patterns of mouse skin.

关 键 词：芥子气 蛋白表达 皮肤 

分 类 号：R114[医药卫生—卫生毒理学]