剪切修复基因xpd抑制pdgf-bb诱导人脐静脉平滑肌细胞增殖的作用及信号机制-effect of xpd on pdgf - bb - induced proliferation of human umbilical vein smooth muscle cells and its signaling mechanism.docxVIP

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剪切修复基因xpd抑制pdgf-bb诱导人脐静脉平滑肌细胞增殖的作用及信号机制-effect of xpd on pdgf - bb - induced proliferation of human umbilical vein smooth muscle cells and its signaling mechanism.docx

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剪切修复基因xpd抑制pdgf-bb诱导人脐静脉平滑肌细胞增殖的作用及信号机制-effect of xpd on pdgf - bb - induced proliferation of human umbilical vein smooth muscle cells and its signaling mechanism

摘要（2） GSK3β 介导 XPD 抑制 PDGF-BB 诱导的 VSMC 增殖；（3） GSK3β 可能通过 CDK4、cyclinD1 介导 XPD 抑制 VSMC 增殖的作用。关键词：剪切修复基因；细胞增殖；血小板源性生长因子；糖原合成激酶 3βABSTRACTOBJECTIVE:To investigate the effects of XPD on vascular smooth muscle cell （VSMC） proliferation induced by platelet-derived growth factors-BB (PDGF-BB ) , and the role of Glycogen synthase kinase-3β (GSK3β) in the Inhibitory Effects of XPD on HUVSMC Proliferation induced by PDGF-BB.METHODS：Human Umbilical Vein Smooth muscle Cell were cultured; Establish the model of PDGF-BB inducing vascular smooth muscle cell proliferation ,and cultivate it under various conditions.Optimize the PDGF-BB concentration and the incubation time. Harvest cells for 24 hours with final PDGF-BB concentration in the medium at 0ng/ml,10 ng/ml,30 ng/ml,50 ng/ml,80 ng/ml. Cell was detected by MTT and the best concentration of PDGF-BB was chosen .Cells were incubated with PDGF-BB at the concentration selected for 0,12,24,36,and 48 hours, followed by MTT detection.Incubate cells with PDGF-BB at the concentration selectedfor 0,1,8,16, 24,32,40and 48 hours, then harvest cells and extract cellular proteins. The expression of XPD protein was detected by Western Blotting.Extract the recombinant plasmid pEGFP-N2 ／ XPD and vacant vectorplasmid . The expression of XPD protein was detected by Western Blotting.Incubate transfected recombinant plasmid pEGFP-N2／XPD and transfected pEGFP-N2 VSMC cells with PDGF-BB. The experiments were divided into five groups:(1) Blank control group;(2)pEGFP-N2 group;(3)pEGFP-N2/XPD group; (4)PDGF-BB group;(5)PDGF-BB+pEGFP-N2 group;(6)PDGF-BB+ pEGFP- N2/XPD group. By means of Western Blotting, the expressions of XPD, GSK3β, p-GSK3β, CDK4,cyclinD1 protein were detected. By means of Flow Cytometry, cell cycle was detected.HUVSMC were transfected with recombinant plasmidpEGFP-N2／XPD,andcultured 24 hours with 0.25μM GSK3β inhibitor. The experiments were divided into four groups:（1）Blank control；（2）DMSO group；(3)pEGFP-N2/XPD group；(4)