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目的考察不同载脂蛋白E基因型存在时,氟伐他汀对L-02细胞3-羟基-3-甲基戊二酰辅酶A(HMG-Co A)还原酶活性的影响。方法建立通过检测还原辅酶(NADPH)反应酶活性的高效液相色谱检测方法及样品处理方法。摸索细胞总蛋白与酶的反应体系,研究氟伐他汀对HMG-Co A还原酶活性的抑制作用。结果建立的高效液相色谱检测方法精密度、准确度及重现性均符合要求。NADPH在2.5~1000.0μmol·L-1线性关系良好。0~5μg·m L-1氟伐他汀对L-02细胞中HMG-Co A还原酶活性的抑制作用成浓度依赖性,最大抑制率为49.73%。载脂蛋白E2、E3、E4分别存在时,氟伐他汀对HMG-Co A还原酶活性的抑制率分别为38.02%,25.12%,21.92%,与野生型E3相比,Apo E2差异有统计学意义(P>0.05)。结论载脂蛋白E不同基因型存在时,氟伐他汀对L-02细胞HMG-Co A还原酶活性抑制作用有差异。
Objective To investigate the effect of fluvastatin on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-Co A) reductase activity in L-02 cells in the presence of different apolipoprotein E genotypes. Methods High performance liquid chromatography (HPLC) detection method and sample processing method were established by detecting the enzyme activity of reducing coenzyme (NADPH). To explore the reaction system of total cellular protein and enzyme to study the inhibitory effect of fluvastatin on HMG-Co A reductase activity. Results The precision, accuracy and reproducibility of the established HPLC method met the requirements. NADPH in 2.5 ~ 1000.0μmol·L-1 good linear relationship. The inhibitory effect of fluvastatin at 0 ~ 5μg · m L -1 on HMG-Co A reductase activity in L-02 cells was concentration-dependent, with the maximum inhibition rate of 49.73%. The inhibitory rates of fluvastatin to HMG-Co A reductase were 38.02%, 25.12% and 21.92% in the presence of apolipoprotein E2, E3 and E4, respectively. Compared with wild type E3, Apo E2 showed statistically significant difference Significance (P> 0.05). Conclusion Fluvastatin can inhibit the HMG-Co A reductase activity in L-02 cells in the presence of different apolipoprotein E genotypes.