摘要
目的探讨血凝素样氧化低密度脂蛋白受体1(LOX-1)在氧化低密度脂蛋白(ox-LDL)活化人脐静脉内皮细胞p38丝裂素活化蛋白激酶(p38MAPK)信号通路中的作用。方法培养人脐静脉内皮细胞(HUVEC),用不同浓度的ox-LDL孵育,通过W estern印迹检测LOX-1、p38MAPK蛋白水平,逆转录聚合酶链法观察LOX-1 mRNA表达,并以LOX-1特异阻断性抗体(JTX92)预处理内皮细胞,检测p38MAPK蛋白水平。四氮唑蓝法观察ox-LDL对细胞活力的影响。结果ox-LDL引起内皮细胞形态结构的改变,而且抑制内皮细胞的生长(P<0.05);ox-LDL呈浓度依赖性地上调LOX-1蛋白和mRNA表达(均P<0.05),并呈浓度依赖性激活p38MAPK通路,不同浓度ox-LDL(25μg/m l,50μg/m l,100μg/m l)组p-p38MAPK蛋白质表达水平均明显高于对照组(48±7,79±15,113±14 vs 24±5,P<0.01);JTX92+ox-LDL(100μg/m l)组p-p38MAPK蛋白水平明显低于ox-LDL(100μg/m l)组,(58±13 vs 113±14,P<0.01)。结论ox-LDL通过LOX-1途径激活p38MAPK信号通路,LOX-1上调是ox-LDL致动脉粥样硬化的重要环节。
Objective To investigate the effects of lecitin-like oxidized low-density lipoprotein receptor-1 （LOX-1） on the p38 mitogen-activated protein kinase （p38MAPK） induced by oxidized low- density lipoprotein （ox-LDL） in endothelial ceils. Methods Human umbilical vein endothelial cells of the line ECV304 were cultured and treated with LDL 25 μg/ml and ox-LDL of 3 different concentrations （ 25 μg/ml, 50 μg/ml, and 100 μg/ml） for 24 hours. ECV304 ceils without treatment of ox-LDL were used as control group. MTT method was used to detect the optical density （OD） of different groups. RT-PCR was used to detect the mRNA expression of LOX-1. Western blotting was used to detect the protein expression of LOX-1. Another ECV304 cells were randomly divided into 7 groups to be treated with LDL （25 μg/ml）, ox- LDL of the concentrations of 25 μg/ml, 50 μg/ml, and 100 μg/ml, ox-LDL 100 μg/ml ＋ JTX92 （ LOX-1 blocking antibody） 10 μg/ml, or JTX92 10μg/ml, or not to be treated with LOX-1 and/or JTX92 （as control group）. The p38MAPK protein expression and phophorylated p38MAPK （p-p38MAPK） protein expression were detected by Western blotting. Results The A values of the ECV304 ceils treated with ox- LDL were significantly lower than that of the control group dose-dependently （ all P 〈 0.05 ）, however, the A value of the LDL group was not significantly different from that of the control group （ P 〉 0.05 ）. The LOX- 1 mRNA expression and protein expression were significantly increased in the ox-LDL groups dosedependently （beth P 〈0.01 ）. However, the LOX-1 mRNA expression and protein expression of the LDL group were not significantly different from those of the control group （ beth P 〉 0.05 ）. The p38MAPK protein expression level was not significantly different among different groups （ all P 〉 0.05 ）. ＂The p- p38MAPK protein expression was up-regnlated by ox-LDL dose -dependently （P 〈 0.05 ）. The p-p38MAPK protein level of the ox-LDL ＋ JTX92 group was s
出处
《中华医学杂志》
CAS
CSCD
北大核心
2006年第24期1697-1700,共4页
National Medical Journal of China
