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目的构建血管平滑肌源性泡沫细胞模型,探讨泡沫细胞形成过程中,酰基辅酶A:胆固醇酰基转移酶1(ACAT1)在转录、表达和酶活性水平上的改变。方法培养小鼠血管平滑肌细胞(VSMC),采用100g/mL氧化低密度脂蛋白(ox-LDL)作用于VSMC72h,建立泡沫细胞模型。油红染色证明泡沫细胞形成。用半定量RT-PCR、Westernblot和放射性同位素标记的方法,分别检测VSMC和泡沫细胞中ACAT1mRNA、蛋白水平及酶活性。结果在VSMC转化为泡沫细胞过程中,ACAT1酶活性上升(P<0.05),同时ACAT1蛋白及mRNA水平约提高了1倍。结论 VSMC转变为泡沫细胞过程中,ACAT1转录水平上调导致ACAT1蛋白表达及其酶活性提高,促使肌源性泡沫细胞的形成。
OBJECTIVE: To construct vascular smooth muscle-derived foam cell model and to investigate the changes of acyl-CoA: cholesterol acyltransferase 1 (ACAT1) transcription, expression and enzyme activity during the formation of foam cells. Methods Vascular smooth muscle cells (VSMCs) were cultured in vitro. Foam cell models were established by using ox-LDL (100g / mL) for 72h. Oil red staining confirmed foam cell formation. Semi-quantitative RT-PCR, Western blot and radioisotope labeling methods were used to detect ACAT1 mRNA, protein level and enzyme activity in VSMC and foam cells respectively. Results The ACAT1 activity increased (P <0.05) during VSMC transformation into foam cells, and the protein and mRNA level of ACAT1 increased by about 1 time. Conclusion The up-regulation of ACAT1 transcription leads to the increase of ACAT1 protein expression and its enzymatic activity during the process of transforming VSMC into foam cells, which leads to the formation of myogenic foam cells.