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为观察高密度脂蛋白是否能拮抗氧化型低密度脂蛋白对血管平滑肌细胞释放一氧化氮的抑制作用,培养了人脐动脉平滑肌细胞并用脂多糖和γ-干扰素诱导一氧化氮的释放。应用GRIESS试剂测定细胞培养基中亚硝酸盐的含量.以免疫组织化学染色和逆转录--聚合酶链反应分析诱导型一氧化氮合酶基因的表达。结果发现,氧化型低密度脂蛋白可使脂多糖和γ-干扰素诱导的平滑肌细胞一氧化氮合酶及其mRNA水平下降、抑制一氧化氮释放。而高密度脂蛋白能拮抗氧化型低密度脂蛋白对平滑队细胞释放一氧化氮的抑制作用。0.25g/L高密度脂蛋白即能显著增加一氧化氮的释放(P<0.05),高密度脂蛋白浓度达到1.5g/L可使得一氧化氮的释放增加4.5倍。结果表明,高密度脂蛋白能提高一氧化氮合酶基因表达水平,促进一氧化氮的释放。高密度脂蛋白的作用具有浓度效应。
To observe whether high-density lipoprotein (LDL) can antagonize the inhibitory effect of oxidized low-density lipoprotein on nitric oxide release from vascular smooth muscle cells, human umbilical artery smooth muscle cells were cultured and the release of nitric oxide was induced by lipopolysaccharide and interferon-γ. The nitrite content in cell culture media was determined using GRIESS reagent. The expression of inducible nitric oxide synthase gene was analyzed by immunohistochemical staining and reverse transcription - polymerase chain reaction. The results showed that oxidized low density lipoprotein could decrease the level of nitric oxide synthase and its mRNA in smooth muscle cells induced by lipopolysaccharide and IFN-γ and inhibit the release of nitric oxide. High density lipoprotein (LDL) can antagonize the inhibitory effect of oxidized low density lipoprotein on the release of nitric oxide from smooth muscle cells. 0.25g / L high density lipoprotein can significantly increase the release of nitric oxide (P <0.05), high density lipoprotein concentration of 1.5g / L can make the release of nitric oxide increased 4.5 times. The results show that high-density lipoprotein can increase nitric oxide synthase gene expression and promote the release of nitric oxide. The role of HDL has a concentration effect.