| [Background]:Cardiovascular disease is the main cause of death in the developed and developing countries. Atherosclerosis(AS) is a chronic multi-factor disease and the risky factors leading to AS have been more than200, which the number is still growing. AS is closely related to cardiovascular disease. Coronary artery AS can lead to coronary heart disease and myocardial infarction, and aortic AS can lead to dissecting aneurysm. AS develops in a progressive way, including three stages:fatty streak formation, fibrous plaque formation and atheromatous plaque formation. Endothelium dysfunction is the early sign of AS, and apoptosis of smooth muscle cell is the late sign of AS. Currently, statins, angiotensin-converting enzyme inhibitors (ACEI) and antiplatelet drugs are the main options for the treatment of AS. The mechanism for the treatment includes reducing blood lipid, alleviating vascular inflammation, protecting and improving the endothelium function, anti-proliferation of smooth muscle cell and inhibiting the platelet functions. Interventions and surgical treatments are considered when patients have serious AS disease. Due to the high rate of morbidity and mortality of AS and the complicated mechanism underlying the development of AS, looking for a new target and a more potent treatment for atherosclerosis seems neccessary. Angiotensin-converting enzyme2(ACE2) is a new member of renin-angiotensin system(RAS). Studies have found that ACE2inhibited the early AS plaque formation through inhibiting vascular smooth muscle proliferation and improve the function of endothelial cells. Current study support the idea that increased expression and improved function of ACE2may probably play an important role in the treatment of AS and other related disease.[Objective]:To construct replication-deficient recombinant adenovirus Ad-ACE2and to investigate the effects of ACE2on the severity of AS in apolipoprotein-E knockout (ApoE-/-) mice.[Methods]:Replication-deficient recombinant adenovirus(Ad-ACE2) was constructed firstly, and high-fat feeding was applied to establish16mice models of atherosclerosis, which were then divided into two groups randomly. Furthermore,8mice receiving normal diet were included for control. The three groups were then received Ad-ACE2, Ad-EGFP and saline by tail vein injection respectively. The size of plaque, the lipid content, macrophage filtration, the protein expression of monocyte chemoattractant protein-1(MCP-1), vascular cell adhesion molecule-1(VCAM-1) and E-selectin were evaluated by HE, Oil Red O stainting, immunohistochemical method and Western blotting; Oxidative stress in each group was also measured by using a kit.[Results]:ACE2gene and pMD18-T plasmid were obtained by digesting the recombinant plasmid pMD18-T-ACE2with EcoRI+Sall enzyme, proving the recombinant plasmid pMD18-T-ACE2was constructed successfully; ACE2gene(2.4Kb) and pDC316plasmid(3.9Kb) were obtained by digesting the shuttle plasmid pDC316-ACE2with EcoRI+Sall enzyme, proving the shuttle plasmid pDC316-ACE2was constructed successfully, and PCR and DNA sequencing results also supported the fact that the shuttle plasmid pDC316-ACE2was constructed successfully; Ad-ACE2was successfully constructed as proved by PCR. ACE2gene and protein expression were found in human umbilical vein endothelial Cell(HUVEC) transfected by Ad-ACE2; The size of plaque, the lipid content, macrophage filtration, MCP-1and the protein expression of VCAM-1and E-selectin were significantly lower in ACE2gene treatment group than in ACE2gene control group; Oxidative stress in ACE2gene treatment group was alleviated compared with in ACE2gene control group.[Conclusion]:Ad-ACE2is constructed successfully. ACE2gene and protein expression in HUVEC were detectable after HUVEC was transfected by Ad-ACE2, Overexpression of ACE2gene can reduce the size of plaque, the lipid content, macrophage filtration, and the protein expression of MCP-1, VCAM-1and E-selectin in the abdominal atherosclerotic plaque, and alleviate the oxidative stress simultaneously, thus alleviate the severity of atherosclerotic plaque in ApoE-/-mice. |
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