人参皂苷Rb1逆转白消安干预人脐静脉内皮细胞NO生成作用及其信号转导机制  

Mechanism of Ginsenoside Rb1 on rerersing busulfan-inhibited nitric oxide synthesis in human umbilical vein endothelial cells via activating PI3K/AKT signaling

作　　者：方峻[1] 陈红霞[1] 谢薇[1] 毛星宁[1] 夏凌辉[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液科,湖北武汉430022

出　　处：《中国医院药学杂志》2008年第20期1761-1764,共4页Chinese Journal of Hospital Pharmacy

摘　　要：目的:研究人参皂苷Rb1对白消安(BU)干预人脐静脉内皮细胞(HUVEC)一氧化氮(NO)合成作用的影响,并进一步探讨此影响的信号转导机制。方法:体外培养HUVEC,以硝酸还原酶法检测各组细胞培养上清液中NO的含量,实时荧光定量PCR(RealTime-PCR)法检测HUVEC中内皮型一氧化氮合酶(eNOS)mRNA表达;以特异性抑制剂LY294002阻断PI3K/AKT信号转导通路,Western blot印迹法检测细胞内磷酸化丝/苏氨酸蛋白激酶AKT(P-AKT)表达。结果:①BU明显降低HUVEC中的eNOSmRNA表达及细胞培养上清中的NO水平;预先用人参皂苷Rb1干预可以拮抗BU对HUVEC的这种影响,提高eNOS mRNA表达及NO合成;②BU明显抑制HUVEC表达P-AKT,Rb1则可增强P-AKT表达,拮抗白消安对P-AKT的抑制作用;③人参皂苷Rb1拮抗白消安抑制内皮细胞NO合成的作用能被LY294002拮抗;40μmol.L-1的LY294002即可明显抑制磷酸化AKT的表达和NO的合成。结论:BU可抑制HUVEC合成NO,人参皂苷Rb1可逆转此作用,其机制与激活PI3K/AKT信号转导通路密切相关。人参皂苷可能籍此机制保护血管内皮细胞免受化疗药物损伤。OBJECTIVE To investigate the effect of ginsenoside Rhl on busulfan-inhibited nitric oxide（NO） synthesis in human umhilical vein endothelial cells（HUVECs） and its signaling mechanism. METHODS HUVECs were cultured. NO levels in the cuhural supernatant were examined by carbonate reductase assay. Endothelial nitric oxide synthase（eNOS） mRNA levels of HUVECs were tested by real time RT-PCR. Ly294002 were used to specifically inhibit PI3K/AKT signal transduction. Expression of phosphorylated AKT were explored by Western bloting. RESULTS （1）Busulfan（BU） significantly decreased the expression of eNOS mRNA in HUVECs and NO level in cultural supernatant. Pre-treatment with ginsenoside Rb1 reversed the BU-inhibited NO synthesis in a dose-dependent manner, resulted in enhanced eNOS mRNA expression and NO level. （2）BU inhibited P-ANT expression while Rbl increased AKT phosphorylation against this effect of BU. （3）Reverse of BU-inhibited NO synthesis by Rbl was depressed by LY 294002. 40μmol·L^-1 Ly294002 significantly blocked gensinoside Rbl-enhanced P-AKT expression and NO synthesis. CONCLUSION BU could inhibit NO synthesis but ginsenoside Rbl could reverse this effect via activating PI3K/ANT signaling in HUVECs. Therefore, ginsenoside Rbl could protect endothelial cells against chemotherapy induced damage through this mechanism.

关 键 词：人参皂苷 一氧化氮 信号通路 人脐静脉内皮细胞 

分 类 号：R969[医药卫生—药理学]