摘要
目的观察急性内皮剥脱损伤对血管壁基质金属蛋白酶（matrix metalloproteinase，MMP）9的活化及其可能的信号调控机制。方法以C57BL／6小鼠为研究对象，实施股动脉内皮剥脱损伤后，分别制备股动脉总蛋白和膜蛋白提取液，用明胶酶谱法检测MMP9活性，蛋白质印迹（Western Blot）检测MMP9抗原表达和膜蛋白中蛋白激酶c（protein kinaseC，PKc）多种异构体的表达水平；并以PKc8抑制剂Ruboxistaurin预处理小鼠72h后，观察急性血管损伤对MMP9表达的影响。结果内皮剥脱损伤4～24h，小鼠股动脉MMP9活性增加，最大诱导表达量在损伤后8h，为对照组的（99．33±9．50）倍（F=51．49，P〈0．01）；MMP9抗原表达与其活性呈现平行变化。内皮剥脱损伤15～120min，股动脉膜蛋白提取液中PKCβⅡ被快速诱导表达，30min时达最大值，为对照组的（7．50±0．60）倍（F=207．06，P〈0．01）。PKCβ抑制剂Ruboxistaurin可减少血管损伤诱导的MMP9活化（F=53．96，P〈0．01）。结论在小鼠股动脉内皮剥脱模型中，MMP9活化至少部分由PKCβ信号所介导，阻断PKCβ信号通路可望成为抑制血管损伤后MMP9过度活化的干预靶点。
Objective To investigate the activation of matrix metalloproteinase （MMP）9 by acute arterial injury and the involved signaling mechanism in murine femoral artery. Methods In the C57BL/6 mice femoral artery denudation injury were performed. Total protein and membrane protein extracts were prepared from targeted arteries. The MMP 9 activity was measured by zymography assay, the expressions of MMP 9 antigen and protein kinase C （PKC） isoforms were measured by Western blot. Seventy-two hours after mice fed with PKCβ inhibitor （ruboxistaurin）, the denudation injury triggered MMP 9 activation was reassessed. Results Within 4-24 hours after denudation injury, MMP 9 activity in femoral arteries was significantly increased, with a peak induction of （99.3± 9.5） times the sham control （F=51.49,P〈0.01） at 8 h. MMP 9 antigen increased in parallel with MMP 9 activity. Within 15-120 minutes after denudation injury, there was a significant induction of PKCβⅡin membrane fraction of femoral arteries, with a maximum induction of （7.50±0.60） times the sham control （F=207.06,P〈0.01）at 30 min. Injury-induced MMP 9 activation was significantly inhibited by ruboxistaurin. Conclusions MMP 9 activation is, at least in part, mediated by PKCβ in acute arterial denudation injury, it highlights the new target for therapeutic intervention to suppress the over-activation of MMP 9, which plays a critical role in restenosis.
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2011年第7期589-592,共4页
Chinese Journal of Geriatrics
基金
福建省医学创新课题（2009CXB-18）
福建省教育厅课题（JA07103）
关键词
蛋白激酶C
基质金属蛋白酶9
内皮
血管
Protein kinase C-delta
Matrix metalloproteinase 9, Endothelium, vascular
