赣南医学院学报2024，Vol.44Issue(1)：1-8,58,9.DOI:10.3969/j.issn.1001-5779.2024.01.001
极光激酶A通过促进巨噬细胞源性泡沫细胞形成参与动脉粥样硬化发生发展
The role of Aurora kinase A in atherosclerosis by promoting macrophage-derived foam cell formation
摘要
Abstract
Objective:To determine whether Aurora kinase A(AurkA)participates in the development of atherosclerosis(AS)by regulating the formation of macrophage-derived foam cells.Methods:Gene expression profile of atherosclerotic lesions from AS mice was downloaded from GEO data(GSE 19286)and analyzed for AurkA expression changes using specific probes.Eight-week-old C57BL/6J and ApoE-/-male mice were divided into normal-diet-fed C57BL/6J control group(CC group),normal-diet-fed ApoE-/-control group(AN group)and high-fat-diet-fed ApoE-/-AS group(AH group).The high-fat-diet contained 21%fat and 0.5%cholesterol.Peripheral blood,aorta,liver and spleen samples were collected at 16-week timepoint,and blood lipid(TC,TG,LDL-C)levels and en face aorta plaque buildup were analyzed to ensure the successful establishment of AS model.Total RNA was extracted from aorta,liver and spleen to assess AurkA mRNA levels using qPCR method.After that,8-week-old ApoE-/-male mice were randomly divided into vehicle-treated control group(MC group)and AurkA inhibitor MLN8237-treated group(M group),and fed with high-fat diet for 16 weeks.Starting at 12-week timepoint,M group mice were orally administrated with MLN8327(20 mg·kg-1,twice per week)for 4 weeks and MC group mice were treated with vehicle(10%2-hydroxypropyl-β-cyclodextrin和1%sodium bicarbonate)following the same routine.The mice of both groups were weighed weekly and tissues including peripheral blood,heart,aorta were excised at week 16.Peripheral blood samples were used to assess blood lipid levels and determine the proportion of myeloid cells using flowcytometry.Aorta samples were fixed for en face oil red staining and heart samples were used to prepare aortic root sections which were then stained with HE and oil red to evaluate the degree of plaque accumulation and lipid deposition.Lastly,the expression pattern of AurkA within different hematopoietic cell lineages were analyzed using bloodspot database and cellular model was constructed to study the role of AurkA during foam cell formation.Bone marrow cells were collected from 8-week-old C57BL/6J mice and stimulated with 10 ng·mL-1 M-CSF to induce bone marrow-derived macrophage cells.The induced cells were replated into control group(Con),ox-LDL-stimulated group(ox-LDL)and AurkA inhibitor-treated group(ox-LDL+TCS),and the latter two groups were stimulated with 100 μg·mL-1 ox-LDL to induce foam cell formation.Meanwhile,ox-LDL+TCS group received 10 nM AurkA inhibitor TCS7010 treatment and ox-LDL group received same amount of vehicle treatment.After 48-hour incubation,oil red staining was performed to assess the lipid accumulation within foam cells.Results:Gene expression profile data showed that AurkA was upregulated in the atherosclerotic lesion area.Compared to either CC group or AN group,the serum levels of TC,TG and LDL-C within AH group were increased(P＜0.05),and there was a substantial accumulation of atherosclerotic plaques in the aorta from AH group mice,indicating the successful construction of AS model.In the AH group,the AurkA mRNA levels were higher in liver,spleen and aorta than that in the AN group(P＜0.05).With the treatment of MLN8237,the body weight,serum lipid levels and aortic total plaque area were not different between MC group and M group(P＞0.05),but the plaque accumulation and its lipid content within the aortic root were decreased for M group compared to MC group(P＜0.05).Flow cytometry data revealed that the myeloid cell percentages in the peripheral blood were comparable between two groups(P＞0.05),but the monocyte proportion was lower in M group than that in MC group(P＜0.05).Expression pattern analysis showed that AurkA is highly expressed in myeloid cells,especially in macrophages.In the cellular experiments,the oil red-positive area was reduced for ox-LDL+TCS group compared to ox-LDL group(P＜0.05),suggesting that AurkA inhibitor decreased lipid accumulation in macrophage and thereby constrained the formation of foam cells.Conclusion:AurkA was overexpressed during the development of atherosclerosis,and it promoted the plaque formation by regulating the number of monocytes in peripheral blood and the generation of macrophage-derived foam cells.Overall,inhibition of AurkA activity effectively limited the progression of atherosclerosis.关键词
极光激酶A/动脉粥样硬化/泡沫细胞形成/斑块生成/髓系细胞生成Key words
Aurora kinase A/Atherosclerosis/Foam cell formation/Plague formation/Myelogenesis分类
医药卫生引用本文复制引用
耿戈戈,张峰华,刘晓炎,樊雯婷,甘滔..极光激酶A通过促进巨噬细胞源性泡沫细胞形成参与动脉粥样硬化发生发展[J].赣南医学院学报,2024,44(1):1-8,58,9.基金项目
国家自然科学基金项目(31960201) （31960201）
江西省教育厅科学技术研究项目(GJJ170861) （GJJ170861）
赣南医学院校级科技创新团队课题(TD201903) （TD201903）
赣南医学院校级重点课题(ZD201702) （ZD201702）
研究生创新创业基金项目(YC2022-X010) （YC2022-X010）
