Establishment Of The Model Of Atherosclerosis In Mice With Lupus Erythematosus And Its Interventional Mechanism

Purposes1. Modelling systemic lupus erythematosus (SLE) accompanied with atherosclerosis in mice.2. Observe the effect of systemic lupus erythematosus (SLE) on atherosclerosis and lipid metabolism in murine models.3. Observe the feature of lupus atherosclerosis, especially in atherosclerotic plaques, and discuss the possible mechanisms behind.4. We also studied the effect of hydroxychloroquine on lupus atherosclerosis and lipid. Furthermore, we analyzed the possible mechanism behind.Methods1. ApoE-/- and C57BL/6 mice were given intraperitoneal injection of 0.5ml pristane to induce lupus. Some mice were treated with hydroxychloroquine (HCQ) through drinking water.2. Onset of lupus was evaluated by measuring anti-dsDNA antibody concentration. Onset of atherosclerosis was measured by aorta oil O staining.3. Mice were sacrificed after 6 months. We examined urine protein. Serum was collected and was tested for lipid, albumin, IgG and interferon-a.4. Some aortas were stained by oil O. Others were digested and stained with CD45+. CD 19+. CD68+、CD11c+ cells and applied for flow cytometry. The roots of aortic valve were sliced and applied for oil O staining, immunofluorescence staining of IgG,CD19+、CD68+、CD11c+cells.5. Spleen single cell suspensions were prepared and were stained with CD3+、 CD 19+、CD11b+、CD11c+ cells for flow cytometry.Results1. All of pristane injected mice presented with elevated dsDNA antibodies. HCQ reduced dsDNA antibody positive rate.2. Part of lupus mice showed unhairing and skin hemorrhage. These phenomenons were restricted to lupus mice.3. ApoE-/- mice were heavier.4. LDL-C and TC values were higher in apoE-/- mice compared to C57BL/6 mice. TC values were lower in lupus mice compared to non-lupus mice. HCQ administration had no effect on lipid level. Serum albumin levels of lupus mice were lower compared to non-lupus ones, while HCQ had no effect on albumin levels.5. Atherosclerotic plaques were larger in lupus mice. Only apoE-/- mice developed atheroma. HCQ inhibited the formation of atheroma.6. The number of total aortic cells, WBCs, macrophages dendritic cells and IgG deposition increased in aortas of lupus mice. B cell ratio, on the other hand, decreased in aortas from lupus mice. HCQ reversed the effect of lupus. HCQ had little effect on non-lupus mice. The localization of macrophages and dendritic cells were almost identical. Serum IgG of lupus mice were higher than non-lupus ones, and serum IgG were negatively correlated with IgG deposition in atheroma.7. Spleen index of lupus mice were higher. Spleen lymphocytes reduced in lupus mice, while HCQ had no significant impact on spleen lymphocytes ratio. Spleen dendritic cells in lupus mice increased, while HCQ had no impact on it.8. Urine protein were higher in lupus mice. HCQ was capable of reducing urine protein of C57BL/6 mice.9. Serum IFN-α level were not changed in lupus mice.Conclusions1. Lupus was successfully induced by pristane injection. HCQ is capable of reducing autoantibodies.2. Atherosclerotic plaques were only observed in apoE-/- mice.3. Lupus accelerated atheroma development.4. Lupus mice had lower lipid profile. On the other hand, lupus atheroma were characterized by inflammatory cell infiltration, such as gathered dendritic cells (DCs), macrophages, IgG and reduced B cells. Decreased lymphocytes and magnified DCs in spleen were also observed in lupus mice. Serum albumin were reduced and IgG were increased in lupus mice.5. HCQ administration inhibited dsDNA antibodies generation and attenuated atherosclerosis.6. HCQ had no effect on lipid profile. HCQ showed the ability to reverse lupus atheroma content.7. High fat diet leaded to elevated lipid and abnormal immune system.8. Serum IFN-a level were not changed in lupus mice.