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采用组织贴壁培养法建立氧化型低密度脂蛋白(ox-LDL)诱导血管平滑肌细胞(VSMC)的增殖模型,以四甲基偶氮唑蓝(MTT)法观察车前子多糖(plantain seed polysaccharide,PSP)对VSMC增殖的影响,结果表明ox-LDL诱导的VSMC增殖明显(P<0.05),认为该增殖模型成功建立。利用比色法测定丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、一氧化氮(NO)含量以及一氧化氮合酶(NOS)的活性,探讨PSP在细胞水平的抗氧化作用,结果显示给予PSP后,MDA含量显著下降,SOD、NO和NOS水平明显升高(P<0.05),表明PSP具有一定的抗氧化作用。RT-PCR检测原癌基因(c-myc)和单核细胞趋化蛋白-1(MCP-1)水平,观察PSP对c-myc和MCP-1mRNA表达的影响,以寻找药物的靶点并探讨PSP抗动脉粥样硬化作用的分子机制。结果表明,PSP下调c-mycmRNA和MCP-1mRNA的表达,可能是抗动脉粥样硬化的机制之一。
The vascular smooth muscle cells (VSMC) proliferation model induced by oxidized low-density lipoprotein (ox-LDL) was established by tissue adherent culture method. Plantain seed polysaccharide was observed by MTT assay. , PSP) on the proliferation of VSMC, the results show that ox-LDL-induced proliferation of VSMC significantly (P <0.05), that the successful establishment of the proliferation model. The malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, nitric oxide (NO) content, and nitric oxide synthase (NOS) activity were measured by colorimetry to investigate the antioxidant activity of PSP at the cellular level. The results showed that after administration of PSP, the content of MDA significantly decreased and the levels of SOD, NO, and NOS increased significantly (P<0.05), indicating that PSP has a certain anti-oxidation effect. RT-PCR was used to detect the levels of c-myc and monocyte chemoattractant protein-1 (MCP-1). The effect of PSP on the expression of c-myc and MCP-1 mRNA was observed to find the target of drug and explore PSP anti-atherosclerosis molecular mechanism. The results showed that down-regulation of c-myc mRNA and MCP-1 mRNA expression by PSP may be one of the mechanisms of anti-atherosclerosis.