Decrescent role of recombinant HSP۶۰ antibody against atherosclerosis in high-cholesterol diet immunized rabbits

Objective(s): Atherosclerosis is the main cause of cardiovascular disease (CVD) which has a key role in the development of coronary artery disease (CAD). Based on clinical studies, HSP۶۰ is the only HSP that can cause atherosclerosis. In this paper, the expression level of HSP۶۰ and the pathogenic degree of its cloned part was investigated in atherosclerosis condition.Materials and Methods:  After the designation of the specific primers for HSP۶۰, PCR was done by the Pfu enzyme. Subsequently, the PCR products were cloned into a prokaryotic expression vector pET-۲۸a. The resultant recombinant vector was transferred in BL۲۱ and purified. Purification of protein was done by the Nickel affinity column. After confirmation of Western blotting and HSP۶۰ protein purification, purified protein concentration was measured by the Bradford method, and purity was analyzed by SDS PAGE ۱۲%. New Zealand rabbits were tested as an animal model. At the next step, the recombinant protein was injected into the animal model that was on a fatty diet.Results: The prokaryotic expression plasmid pET۲۸a-hps۶۰ was successfully constructed, the HSP۶۰ protein was expressed and purified in Escherichia coli BL۲۱ (DE۳). We found that the rabbit that was receiving the recombinant vaccine with the fatty diet showed a lower amount of fat deposition at the media endothelial level than the rabbit which received only the fatty diet. Conclusion: Taking recombinant protein concomitant with a fatty diet, causes betterment of atherosclerosis via decreasing aggregation of cholesterol and thickness of the endothelial media.

Keywords:

Atherosclerosis , Coronary Artery Disease , Heat- shock proteins , Plasmids , Recombinant protein