Lipoprotein Receptor Activity To Regulate Agents Screening Model And Applied Research

Atherosclerosis is the disease of the highest mortality in many countries. The elevation of serum cholesterol especially LDL-c levels is one of the most relative risk factors that cause atherosclerosis. LDL can be modified by many kinds of cells in the arterial wall. Receptor-mediated endocytosis of modified LDL by macrophages has been implicated in the pathogenesis of atherosclerosis.Research results demonstrate that SR-A and CD36 are responsible for the preponderance of modified LDL uptake in macrophages. Thus, inhibition abilty of SR-A and CD36 to scavenge modified LDL will provide atheroprotection in invo.The majority of LDL-c are taken up and cleared by hepatic LDLR. Upregulation of hepatic LDLR is an effective method to lower serum LDL-c levels. In our anti-atherosclerosis research, aimed at these important receptors tightly concerned with atherosclerotic progression, our research is described as below. 1. Establishment and application of a novel screening model for human scavenger receptor CD36 antagonist:With the RNA of human macrophage U937 as template, the complementary DNA (cDNA) fragment encoding human CD36 was amplified by RT-PCR. The sequence cloned CD36 cDNA was the same as the known sequence. The recombinant transfer vector pVL1393-CD36 was constructed and was co-transfected Sf9 with the baculovirus genome. The recombinant CD36 baculovirus could be harvested from the supernatant of the transfected Sf9 cells 96 hours post-transfection. Purified and amplified the recombinant baculovirus. Compared with Sf9 co-transfected with the transfer vector pVL1393 and the baculovirus genome (mock group), Sf9 cells transfected with recombinant baculovirus expressed CD36 protein detected by Western blot and Flowcytometry. Ligand-receptor binding assay showed that CD36 had been expressed effectively on Sf9 cells and had ligand-binding activities.A high-throughput screening model for CD36 antagonist was established after...