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为了观察氧化型及正常极低密度脂蛋白对血管平滑肌细胞内脂类堆积的不同影响,探索平滑肌细胞摄取氧化型极低密度脂蛋白的受体途径,用200mg/L的氧化型及正常极低密度脂蛋白与猪主动脉平滑肌细胞温育48h后,细胞内甘油三酯含量分别为对照组的3.2倍和10.6倍(P〈0.01),且两个实验组之间相比,差异也有显著性意义(P<0.01);细胞内总胆固醇明显升高(P<0.05).但是两个实验组之间无显著差异(P>0.05)。荧光素异硫氰酸酯标记的氧化型极低密度脂蛋白结合试验表明;平滑队细胞可通过低亲和力受体途径可饱和性地摄取完整的氧化型极低密度脂蛋白(Kd=48.82mg/L,Bmax=2067μg/g)。与正常极低密度脂蛋白相比(Kd=37.17mg/L,Bmax=2596μg/g),Kd值升高,Bmax值减小。竞争抑制实验表明,氧化型极低密度脂蛋白、正常极低密度脂蛋白及正常低密度脂蛋白均对荧光素异硫氰酸酯标记的氧化型极低密度脂蛋白的结合摄取表现出明显的竞争作用(50%),而乙酰化低密度脂蛋白仅能抑制10%。提示氧化型极低密度脂蛋白通过低密度脂蛋白受体和/或极低密度脂蛋白受体途径进入平滑肌细胞。
In order to observe the different effects of oxidized and normal very-low-density lipoproteins on lipids accumulation in vascular smooth muscle cells and to explore the uptake of oxidized very low density lipoprotein receptor by smooth muscle cells, 200mg / L oxidative and normal low The density of lipoprotein and porcine aortic smooth muscle cells after 48h incubation, intracellular triglyceride content were 3.2 times and 10.6 times the control group (P <0.01), and between the two experimental groups The difference was also significant (P <0.01); intracellular total cholesterol was significantly increased (P <0.05). However, there was no significant difference between the two experimental groups (P> 0.05). Fluorescein isothiocyanate labeled oxidized very-low density lipoprotein binding assay showed that smooth cell can uptake intact oxidized very-low density lipoprotein (Kd = 48.82 mg / L, Bmax = 2067 μg / g). Compared with normal very-low-density lipoprotein (Kd = 37.17 mg / L, Bmax = 2596 μg / g), the Kd value increased and the Bmax value decreased. Competitive inhibition experiments showed that the combination of oxidized very-low-density lipoprotein, normal very-low-density lipoprotein and normal low-density lipoprotein showed a significant uptake of fluorescein isothiocyanate labeled oxidized very-low density lipoprotein Competition (50%), while acetylated LDL only inhibited 10%. It is suggested that oxidized very-low-density lipoproteins enter the smooth muscle cells through the low-density lipoprotein receptor and / or the very-low-density lipoprotein receptor pathway.