Madasamy Shanmugavel, Liu David, Lundry Jason, Alderete Benjamin, Kong Raymond, Robinson J Paul, Wu Alan H B, Amento Edward P
Plaxgen Inc;
Plaxgen Inc.
J Vis Exp. 2017 Nov 10(129):56596. doi: 10.3791/56596.
Treatment of dyslipidemia patients with lipid-lowering drugs leads to a significant reduction in low-density lipoproteins (LDL) level and a low to moderate level of increase in high-density lipoprotein (HDL) cholesterol in plasma. However, a possible role of these drugs in altering morphology and distribution of cholesterol particles is poorly understood. Here, we describe the in vitro evaluation of lipid-lowering drug effects in modulating morphological features of cholesterol particles using the plaque array method in combination with imaging flow cytometry. Image analyses of the cholesterol particles indicated that lovastatin, simvastatin, ezetimibe, and atorvastatin induce the formation of both globular and linear strand-shaped particles, whereas niacin, fibrates, fluvastatin, and rosuvastatin induce the formation of only globular-shaped particles. Next, purified very low-density lipoprotein (VLDL) and LDL particles incubated with these drugs showed changes in the morphology and image texture of cholesterol particles subpopulations. Furthermore, screening of 50 serum samples revealed the presence of a higher level of linear shaped HDL cholesterol particles in subjects with dyslipidemia (mean of 18.3%) compared to the age-matched normal (mean of 11.1%) samples. We also observed considerable variations in lipid-lowering drug effects on reducing linear shaped LDL and HDL cholesterol particles formation in serum samples. These findings indicate that lipid-lowering drugs, in addition to their cell-mediated hypolipidemic effects, may directly modulate morphology of cholesterol particles by a non-enzymatic mechanism of action. The outcomes of these results have potential to inform diagnosis of atherosclerosis and predict optimal lipid-lowering therapy.
用降脂药物治疗血脂异常患者可显著降低低密度脂蛋白（LDL）水平，并使血浆中高密度脂蛋白（HDL）胆固醇有低至中度的升高。然而，这些药物在改变胆固醇颗粒形态和分布方面的潜在作用却鲜为人知。在此，我们描述了使用噬菌斑阵列法结合成像流式细胞术对降脂药物在体外调节胆固醇颗粒形态特征方面的作用进行的评估。对胆固醇颗粒的图像分析表明，洛伐他汀、辛伐他汀、依折麦布和阿托伐他汀可诱导球状和线性链状颗粒的形成，而烟酸、贝特类药物、氟伐他汀和瑞舒伐他汀仅诱导球状颗粒的形成。接下来，用这些药物孵育纯化的极低密度脂蛋白（VLDL）和LDL颗粒，结果显示胆固醇颗粒亚群的形态和图像纹理发生了变化。此外，对50份血清样本的筛查显示，与年龄匹配的正常样本（平均11.1%）相比，血脂异常患者中线性形状的HDL胆固醇颗粒水平更高（平均18.3%）。我们还观察到降脂药物在降低血清样本中线性形状的LDL和HDL胆固醇颗粒形成方面的效果存在相当大的差异。这些发现表明，降脂药物除了具有细胞介导的降血脂作用外，还可能通过非酶促作用机制直接调节胆固醇颗粒的形态。这些结果可能为动脉粥样硬化的诊断和预测最佳降脂治疗提供依据。
