人参总皂苷对PDGF-BB所致血管平滑肌细胞增殖周期的影响  

Effect of total ginsenosides on the cell cycle of rat vascular smooth muscle cell proliferation induced by PDGF-BB

作　　者：黄警[1] 黄燮南[1] 张纾[1] 杨丹莉[1] 吴芹[1] 邓江[1] 高杨[1] 

机构地区：[1]遵义医学院药理学教研室暨贵州省基础药理重点实验室,贵州遵义563003

出　　处：《中国药理学通报》2010年第6期787-791,共5页Chinese Pharmacological Bulletin

基　　金：贵州省优秀科技教育人才省长基金项目〔No.黔省合字(2007)53号〕

摘　　要：目的研究人参总皂苷(totel ginsenosides,TG)对血小板源性生长因子BB型(PDGF-BB)所致血管平滑肌细胞(VSMC)增殖周期的影响并探讨其可能的机制。方法组织块贴壁法培养SD大鼠胸主动脉平滑肌细胞,MTT比色法观察TG(10、30、100mg·L-1)对PDGF-BB(25μg·L-1)所致的VSMC增殖的影响;流式细胞仪分析细胞增殖周期;Re-al-timeRT-PCR检测VSMC中内皮型一氧化氮合酶(eNOS)、周期蛋白依赖性蛋白激酶抑制因子P27(KIP1)、原癌基因c-fos、周期蛋白D1(CyclinD1)mRNA的表达。结果在正常细胞中加入TG100mg·L-1不影响细胞的吸光度值及G0/G1期、G2/M期、S期细胞比例(P>0.05);PDGF-BB可明显升高吸光度值(P<0.01),增加S期细胞比例而降低G0/G1期细胞比例(P<0.01),并明显增加c-fos、CyclinD1 mRNA的表达,下调eNOS和P27(KIP1)的表达(P<0.01)。TG低、中、高浓度均明显抑制PDGF-BB诱导的吸光度值升高(P<0.01),降低S期细胞比例而升高G0/G1期细胞比例,明显下调PDGF-BB所致的c-fos及CyclinD1 mRNA高表达(P<0.01),并使eNOS mRNA表达升高(P<0.05),但对P27(KIP1)的表达无明显影响(P>0.05)。结论 TG通过阻止VSMC由G0/G1期进入S期而抑制PDGF-BB所致的增殖,其作用机制可能与其提高eNOSmRNA表达、降低c-fos和CyclinD1 mRNA高表达有关。Aim To observe the effect of total ginsenosides（TG） on the cell cycle of rat vascular smooth muscle cell（VSMC） proliferation induced by platelet-derived growth factor-BB（PDGF-BB） and to probe especially into its mechanism. Methods VSMCs from the thoracic aorta of SD rats were cultured by tissue explant method. Effects of TG （10 mg·L^-1,30 mg·L^-1,100 mg·L^-1） on PDGF-BB-induced VSMC proliferation were evaluated by MTT assay and the cell cycle was analyzed by flow cytometry. Expressions of endothelial NO synthase （eNOS）,proto-oncogene c-fos （c-fos）,CyclinD1 and P27 （KIP1） mRNA in VSMCs were detected by real-time quantitative reverse transcription-polymerase chain reaction（Real-time RT-PCR）. Results TG 100 mg·L^-1 had no effect on the growth and proliferation of normal VSMCs. PDGF-BB could significantly increase the absorbance of VSMCs in MTT assay （P0.01） and the percentage of S phase cells,and degrade the G0/G1 phase cell percentage in the cell cycle （P0.01）. At the same time,PDGF-BB could up-regulate c-fos and CyclinD1 mRNA expressions,and down-regulate eNOS,P27（KIP1） mRNA expressions （P0.01）. Addition of TG （10 mg·L^-1,30 mg·L^-1,100 mg·L^-1） markedly inhibited the PDGF-BB-induced proliferation of the VSMCs,decreased the S phase cell percentage and upgraded the G0/G1 phase cell percentage in the cell cycle; TG could also depress the elevated expressions of c-fos and CyclinD1 mRNA induced by PDGF and up-regulate the expression of eNOS mRNA,but it had no effect on the mRNA level of P27（KIP1）. Conclusions TG could inhibit the VSMC proliferation induced by PDGF-BB through preventing the transformation of the G0/G1 phase cell to S-phase cell in the cell cycle. The mechanisms may be related to its up-regulatory effect on eNOS mRNA and inhibition on C-fos and CyclinD1 mRNA expressions.

关 键 词：人参总皂苷 血管平滑肌细胞 增殖 细胞周期 内皮型一氧化氮合酶 原癌基因C-FOS 血小板源性生长因子 

分 类 号：R-332[医药卫生] R284.1