血小板活化因子对大鼠肺微血管内皮细胞src抑制蛋白激酶c底物基因表达影响-effect of platelet activating factor on src inhibitor protein kinase c substrate gene expression in rat pulmonary microvascular endothelial cells.docxVIP

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血小板活化因子对大鼠肺微血管内皮细胞src抑制蛋白激酶c底物基因表达影响-effect of platelet activating factor on src inhibitor protein kinase c substrate gene expression in rat pulmonary microvascular endothelial cells.docx

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血小板活化因子对大鼠肺微血管内皮细胞src抑制蛋白激酶c底物基因表达影响-effect of platelet activating factor on src inhibitor protein kinase c substrate gene expression in rat pulmonary microvascular endothelial cells

approach using digoxin-labelled oligonucleotide probes. 3. Normal RPMVEC expressed SSeCKS mRNA at a low level, which appeared throughout the cytoplasm with specific hybridization signals. 1.5 h of 10-10, 10-9, 10-8, 10-7 mol/L PAF incubation induced a progressive increase in SSeCKS mRNA expression. When compared SSeCKS mRNA’s mean optical density with the normal control group, difference in each group had statistically significance. Within 0.5, 1.5, 3, 6, 12, 24 h of 10-7 mol/L PAF challenge, the level of SSeCKS mRNA expression markedly raised at 0.5 h, peaked at 1.5 h, then began to decline gradually, and still persisted at a higher level than the normal control group until 24 h. 4. Pre-incubation of 10 μmol/L pyrrolidine dithiocarbamate(PDTC) that inhibits activity of nuclear factor-kappa B(NF-κB) in RPMVECs caused a conspicuous attenuation of PAF- or LPS-induced SSeCKS mRNA expression, whereas no change was found by pretreatment of protein kinase C(PKC) inhibitor bis-indolylmaleimide(BIM).Conclusion: 1. We have successfully cultured and demonstrated the primary RPMVEC in vitro. 2. Through the combination of ISH and computer image analysis software ，We have successfully detected the change of SSeCKS gene expression under differentconditions in RPMVEC. 3. PAF can up regulate expression of SSeCKS mRNA in a dose- and time-dependent manner in RPMVECs. 4. It is NF-κB rather than PKC signal pathway that involve in modulation of the intracellular signaling process that induce SSeCKS mRNA expression.Key words: Platelet-activating factor; Vascular endothelial cell; Lung; Inflammation; Src-suppressed C kinase substrate; Rat; In situ hybridization英文缩略词表Abbreviation英文缩写ALI英文全名acute lung injury中文全名急性肺损伤ARDS BIM BSA DEPC DMEM DNAFBSacute respiratory distress syndrome bis-indolylmaleimidebovine serum albumin diethyl pyrocarbonatedulbeccos modified eagles medium deoxyribonucleic acidfetal bovine serum急性呼吸窘迫综合征双吲哚基顺丁烯二酰亚胺牛血清白蛋白二乙基焦磷酰胺改良 Eagle 氏培养基