Caveolin-1表达对血管平滑肌细胞胆固醇逆转运的调节作用
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国家自然科学基金 (3 0 1710 84)资助;国家 973(G2 0 0 0 0 5 690 5 )资助


Influence of Caveolin-1 on Cholesterol Efflux in Vascular Smooth Muscle Cells
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    摘要:

    通过观察天然和氧化型低密度脂蛋白对细胞内胆固醇转运蛋白caveolin1表达的影响,探索动脉粥样硬化发生过程中血管平滑肌细胞胆固醇逆转运的分子机制。运用Western印迹、逆转录—聚合酶链反应等方法观察天然和氧化修饰的低密度脂蛋白对血管平滑肌细胞caveolin1表达的影响及其差异;运用反义寡核苷酸技术和高效液相色谱、放射性胆固醇标记法等方法初步判断下调caveolin1表达对血管平滑肌细胞胆固醇流出的影响;运用外源基因导入技术观察caveolin1过度表达对氧化型低密度脂蛋白诱导血管平滑肌细胞内胆固醇聚集的调节作用。结果发现:①以正常的未修饰低密度脂蛋白(50mgL)与血管平滑肌细胞孵育96h,caveolin1表达增加60.2%±3.9%,胆固醇流出率为81.7%±4.3%;而在相同量的氧化型低密度脂蛋白刺激下,caveolin1表达减少了54.7%±5.8%,胆固醇流出率下降至26.8%±5.1%。②caveolin1反义寡核苷酸预处理血管平滑肌细胞,再予50mgL的正常低密度脂蛋白处理,细胞胆固醇流出率下降至45.3%±7.3%。③抗氧化剂普罗布考显著对抗氧化型低密度脂蛋白对血管平滑肌细胞caveolin1表达的抑制作用。④外源导入caveolin1表达质粒显著促进氧化型低密度脂蛋白损伤后的细胞胆固醇转运,抑制细胞内胆固醇聚集,细胞总胆固醇含量由175.7±8.9mgg蛋白下降至88.6±3.8mgg蛋白。上述结果提示,caveolin1介导的细胞胆固醇转运和流出是调节血管平滑肌细胞胆固醇平衡的重要机制;氧化型低密度脂蛋白抑制血管平滑肌细胞caveolin1表达,可能是其导致细胞胆固醇代谢障碍、诱导平滑肌源性泡沫化细胞形成的分子机制之一。

    Abstract:

    Aim to study the role of caveolin 1 in vascular smooth muscle cells (VSMC) cholesterol homeostasis. Method Western blot and RT PCR was performed to evaluate the effects of native low density lipoprotein (n LDL) or oxidized low density lipoprotein (ox LDL) on expression of caveolin 1 in VSMC cellular cholesterol efflux, cholesterol content and lipid droplets of VSMC were determined by using cholesterol detection, HPLC and nile red stain respectively Results The results showed that n LDL (50 mg/L) increased caveolin 1 expression by 60.2%±3.9%, and promoted cholesterol efflux rate to 81.7%±4.3%. However, ox LDL (50 mg/L) downregulated caveolin 1 abundance by 54.7%±5.8% and deduced cholesterol efflux rate to 26.8%±5.1%. In addition, the cholesterol efflux rate also significantly decreased when VSMC harboring caveolin 1 antisence. Caveolin 1 overexpression by transfected Pcl neo cav 1 in VSMC results a dramatically decrease in total cellular cholesterol content and lipid droplets induced by ox LDL (50 mg/L). Conclusions Caveolin 1 mediates cellular cholesterol efflux in vascular smooth muscle cells, which is an important mechanism of regulating cellular cholesterol balance. Dowmregulation caveolin 1 expression is involved in foam cell formation derived from vascular smooth muscle cell induced by ox LDL.

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严鹏科,廖端芳,杨永宗. Caveolin-1表达对血管平滑肌细胞胆固醇逆转运的调节作用[J].中国动脉硬化杂志,2002,10(5):379~382.

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  • 收稿日期:2002-08-23
  • 最后修改日期:2002-09-20
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