摘要
目的构建高表达磷脂转运蛋白的细胞模型,探讨半枝莲总黄酮对磷脂转运蛋白(PLTP)表达的影响。方法利用RT-PCR方法扩增磷脂转运蛋白(PLTP)cDNA片段,将其克隆到p3XFLAG-CMV-14载体,得到重组载体PLTP/p3XFLAG-CMV-14,转染于人肝癌细胞系Bel-7402。以不同浓度的半枝莲总黄酮作用于重组细胞,用Western blotting和免疫细胞化学方法检测细胞中PLTP的表达变化。结果转染的人肝癌细胞系Bel-7402细胞模型中PLTP的表达显著增强;与模型(转染于人肝癌细胞系Bel-7402)组比较,终浓度为160 mg/L的半枝莲总黄酮干预Bel-7402细胞24h后,差异有显著性(P<0.05);48h时差异非常显著(P<0.01);72h时差异极显著(P<0.001)。与阳性对照组比较,半枝莲总黄酮高、中、低3个剂量下,作用24、48、72h均无显著性差异(P>0.05)。结论应用RT-PCR、基因重组技术构建了基因重组细胞模型Bel-7402;半枝莲总黄酮可显著抑制重组Bel-7402细胞PLTP的表达。
Objective To construct recombinant phospholipid transfer protein（PLTP） cell model and evaluate the effect mechanisms of Barbata flavonoids （BF） , through the test of their suppressive function of PLTP in Bel-7402 ceils. Methods PLTP cDNA flag was amplified with reverse transcription-polymerase chain reaction （RT-PCR）, PLTP/ p3XFLAG-CMV-14 was constructed and transfected into Bel-7402 cells. The Bel-7402 cells influenced by Barbata flavonoids were analyzed by immunohistochemical assay. Result The expression of PLTP was decreased in Bel-7402 cells treated with 160 mg/L Barbata flavonoids after 24hours,48hours,72hours as compared with the controls （P 〈 0. 05, P 〈0. 05 ,P 〈0.05）. Under high, middle, low-dose BF, treated after 24hours,48hours,72hours, the expression of PLTP had no significant difference as compared with model group（P 〉 0.05）. Conclusion We used gene recombination technology to build recombinant gene cell Bel-7402 . Barbata flavonoids can suppress expression of PLTP in recombinant Bel-7402 cells.
出处
《解剖学报》
CAS
CSCD
北大核心
2011年第2期185-189,共5页
Acta Anatomica Sinica
基金
教育部博士点基金资助项目(20093250120007)
江苏省普通高校研究生科研创新项目(CX09B_320Z)
