低密度脂蛋白和氧化型低密度脂蛋白对HepG2细胞肝脂酶活性及肝脂酶mRNA表达的影响
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Effect of Low Density Lipoprotein and Oxidized Low Density Lipoprotein on Activity and mRNA Expression of Hepatic Lipase in HepG2 Cells
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    摘要:

    为观察低密度脂蛋白和氧化型低密度脂蛋白对肝脂酶合成和肝脂酶活性的影响,将不同浓度的低密度脂蛋白、氧化型低密度脂蛋白与人肝癌细胞株HepG2细胞共同培养,用MTT法测定细胞增殖抑制率,用组织化学法测定细胞肝脂酶活性,用逆转录聚合酶链反应测定细胞肝脂酶mRNA的表达水平。结果发现,低密度脂蛋白浓度在2 7.4 7、5 4 .95mg/L时HepG2细胞逆转录聚合酶链反应产物的吸光度值分别是细胞对照的2 .8倍和2 .3倍,浓度在10 9.89mg/L时逆转录聚合酶链反应产物的吸光度值与细胞对照比较降低2 0 %。细胞肝脂酶活性随低密度脂蛋白浓度增加而降低,两者呈负相关(r=- 0 .95 6 14 ,P<0 .0 5 ) ;低密度脂蛋白浓度在5 4 .95mg/L时开始产生细胞增殖抑制作用,其细胞增殖抑制率与低密度脂蛋白浓度呈正相关(r=0 .91199,P<0 .0 5 )。不同浓度的氧化型低密度脂蛋白处理HepG2细胞其肝脂酶mRNA逆转录聚合酶链反应产物的吸光度值均低于细胞对照孔;脂蛋白浓度为2 7.4 7、5 4 .95及10 9.89mg/L时低密度脂蛋白组细胞肝脂酶活性是氧化型低密度脂蛋白组的3~4倍;浓度在5 4 .95mg/L时氧化型低密度脂蛋白对细胞的增殖抑制作用是低密度脂蛋白的2倍。结果提示,低密度脂蛋白在一定浓度范围能诱导HepG2细胞肝脂酶mRNA表达上调;高浓度低密度脂蛋白及各种浓度氧化型低密度脂蛋白均表现抑制HepG2细胞肝脂酶活性及肝脂酶mRNA表达。

    Abstract:

    Aim To evaluate the effect of LDL and ox LDL on the activity and mRNA expression of hepatic lipase (HL) in HepG2 cells. Methods Gradient concentrations (1.7~1 758 mg/L) of LDL or ox LDL was added into the medium and coincubated with HepG2 cell. The cytostasis rate of HepG2 cell was estimated by MTT and the activity and mRNA expression of HL in these cells were measured with cytochemistry and RT PCR respectively. Results In the 27.47 mg/L and 54.95 mg/L LDL groups, the mRNA expression of HL was up regulating 2.8 times and 2.3 times respectively to the cell control. In the 109.8 mg/L LDL, the mRNA expression of HL decreased 20% to the cell control. The HL activity in HepG2 cells incubated with LDL was negative related with the LDL concentration (r=-0.95614, p<0.05). The cytostasis rate in LDL increased by dose dependent model (r=0.91199, p<0.05). In the ox LDL group, the mRNA expression of HL was inhibited at all concentrations. In 27.47 mg/L and 54.95 mg/L ox LDL, the activity of HL was 3~4 times as much as that of LDL groups. Moreover, at the same concentration of 54.95 mg/L, the cytostasis rate in the ox LDL group was one time higher than in the LDL group. Conclusions At the appropriate range of concentration, LDL can induce the up regulation of mRNA expression of HL. High concentration of LDL and all concentration of ox LDL can inhibit not only mRNA expression but the activity of HL in HepG2 cells.

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张晓刚,陈运贞.低密度脂蛋白和氧化型低密度脂蛋白对HepG2细胞肝脂酶活性及肝脂酶mRNA表达的影响[J].中国动脉硬化杂志,2003,11(2):147~150.

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  • 收稿日期:2002-07-01
  • 最后修改日期:2002-12-19
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