【摘要】目的：观察蛋白激酶C(protein kinase C，PKC)在内毒素/脂多糖(lipopolysacchride， LPS)诱导血管内皮细胞生长因子(vascular endothelial growth factor, VEGF)表达上调中可能发挥的作用。方法：以肺微血管内皮细胞为研究对象，用PKC抑制剂预处理内皮细胞，以RT-PCR检测VEGF mRNA水平的表达变化;Western Blot检测其蛋白质水平的表达变化。结果：(1)VEGF mRNA的表达水平与LPS呈剂量和时间依赖的关系;(2)PKC抑制剂calphostin c预处理内皮细胞能显著抑制LPS诱导VEGF mRNA和蛋白的表达。结论：LPS可能通过PKC信号通路而诱导肺微血管内皮细胞VEGF表达上调。
【关键词】 脂多糖 蛋白激酶C 肺血管内皮细胞 血管内皮细胞生长因子
Protein kinase C mediates lipopolysacchride-induced VEGF gene expression in rat pulmonary microvascular endothelial cells
GE Haiyan, FENG Jian, BEN Suqin, et al (Department of Respiratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001)
[Abstract] Objective: To investigate whether lipopolysacchride(LPS)-induced vascular endothelial growth factor (VEGF) expression was mediated by protein kinase C(PKC). Methods: The rat pulmonary microvascular endothelial cells (RPMVECs) were challenged with LPS before PKC inhibitor(calphostin c) or vehicle, the levels of VEGF mRNA were assayed by RT- PCR, and the levels of VEGF protein were assayed by Western Blot. Results: LPS induced VEGF expression at time and dose dependent manner in RPMVEC. Pretreatment of these cells with the PKC inhibitor(calphostin c) markedly inhibited the LPS-induced VEGF expression. Conclusion: LPS may induce VEGF expression in RPMVEC mediated in part by PKC signal pathway.
[Key words] Lipopolysaccharide; Protein kinase C; Pulmonary microvascular endothelial cell ; Vascular endothelial growth factor血管内皮细胞(vascular endothelial cell, VEC)
处在血液与血管平滑肌细胞之间,能分泌多种生理活性物质,调节血管功能，血管内皮细胞生长因子(vascular endothelial growth factor, VEGF) 即为其中之一。VEGF 具有促进VEC分裂、诱发新血管形成及增加血管通透性的作用[1,2]。本文以培养的肺微血管内皮细胞为模型, 以细菌脂多糖(lipopolysacchride, LPS) 为诱导因素,从VEGF和蛋白激酶C(protein kinase c, PKC)着手，以阐明其发生机制。
1 材料和方法
1.1 材料 LPS(Escherichia coli,0111:B4), calphostin C均购自Sigma公司，DMEM培养基和Trizol 试剂购自Gibco BRL公司，小牛血清购自Hyclone公司，细胞培养板购自Corning公司，VEGF多克隆抗体购自Santa Cruz公司，ECL
[1] [2] [3] [4] 下一页
您现在的位置： 绿色健康网 >> 医学论文 >> 内科论文 >> 正文