circ_0084043/miR-31-5p调节HMGA1参与动脉粥样硬化作用的机制研究  

circ_0084043/miR-31-5p Is Involved in Atherosclerosis by Regulating HMGA1:Investigation of the Mechanisms

作　　者：王克华 李颖[1] 任小璐[1] 王磊[1] WANG Kehua;LI Ying;REN Xiaolu;WANG Lei(General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院,银川750004

出　　处：《四川大学学报(医学版)》2024年第6期1533-1542,共10页Journal of Sichuan University(Medical Sciences)

基　　金：宁夏医科大学2021年校级科研项目(No.XM2021001)资助。

摘　　要：目的探究circ_0084043/miR-31-5p调节HMGA1参与动脉粥样硬化(atherosclerosis,AS)作用的具体机制。方法收集2020年9月–2023年9月49例AS患者的颈动脉斑块及血管组织与对应斑块旁正常组织,qRT-PCR检测circ_0084043、miR-31-5p和HMGA1表达。通过CCK-8、克隆形成、划痕、Transwell实验检测氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导人血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖、迁移、侵袭能力,通过生物信息学、双荧光素酶报告基因检测验证circ_0084043、miR-31-5p、HMGA1之间的靶向调节关系。建立AS小鼠模型,通过油红O、HE染色观察主动脉病变情况,生化检测静脉血血脂指标,qRT-PCR检测miR-31-5p、HMGA1表达情况,免疫组化检测主动脉HMGA1、α-SMA表达情况。结果AS患者斑块、病变血管组织circ_0084043、HMGA1表达水平升高(P<0.05),miR-31-5p表达水平下降(P<0.05)。circ_0084043与miR-31-5p表达水平呈负相关(r=-0.3855,P=0.0062),HMGA1与circ_0084043表达水平呈正相关(r=0.3317,P=0.0199),与miR-31-5p表达水平呈负相关(r=-0.3351,P=0.0186)。miR-31-5p与circ_0084043、HMGA1存在靶向结合位点。与对照组相比,sh-NC组、scramble组、circ_0084043+miR-31-5p mimics组、miR-31-5p inhibitor+sh-HMGA1组、circ_0084043+sh-HMGA1组增殖、克隆形成、迁移、侵袭能力升高(P<0.05);与sh-NC组相比,sh-0084043组则降低(P<0.05);miR-31-5p mimics组、sh-HMGA1组较scramble组增殖、克隆形成、迁移、侵袭能力下降(P<0.05)。与sh-NC组相比,sh-0084043组主动脉脂质斑块、坏死核心面积减小(P<0.05),TC、TG、LDL-C、HMGA1 mRNA及蛋白、α-SMA表达水平下降(P<0.05),HDL-C、miR-31-5p表达水平上升(P<0.05)。结论circ_0084043通过抑制miR-31-5p提高HMGA1表达促进VSMCs增殖、迁移,推动AS进展。Objective To explore the specific mechanism by which circ_0084043/miR-31-5p regulates high mobility group AT-hook 1(HMGA1)and thus participates in atherosclerosis(AS).Methods Carotid plaque tissues,diseased vascular tissue,and corresponding normal tissue from areas adjacent to the plaques were collected from 49 AS patients between September 2020 and September 2023.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to determine circ_0084043 expression.The proliferation,migration,and invasion of vascular smooth muscle cells(VSMCs)induced by oxidized low-density lipoprotein(ox-LDL)were determined by CCK-8,clone formation,scratch,and Transwell assays.The targeted regulatory relationship among circ_0084043,miR-31-5p,and HMGA1 was verified by bioinformatics and dual luciferase reporter gene assay.An AS mouse model was established,and the pathological lesions in the aorta of AS model mice were observed by oil red O and HE stainings.Biochemical testing was performed to assess blood lipids in venous blood.qRT-PCR was used to determine the expression of miR-31-5p and HMGA1.Immunohistochemistry was performed to assess the expression of HMGA1 andα-smooth muscle actin(α-SMA)in the aorta of the AS model mice.Results In AS patients,the expression levels of circ_0084043 and HMGA1 in carotid plaque tissues and diseased vascular tissues were elevated(P<0.05),while the expression level of miR-31-5p was decreased(P<0.05).The expression of circ_0084043 was negatively correlated with the expression of miR-31-5p(r=-0.3855,P=0.0062).HMGA1 expression was positively correlated with circ_0084043 expression(r=0.3317,P=0.0199),and negatively correlated with miR-31-5p(r=-0.3351,P=0.0186).MiR-31-5p had target binding sites for both circ_0084043 and HMGA1.Compared with the control group,the proliferation,clone formation,migration,and invasion abilities of the sh-NC group,the scramble group,the circ_0084043+miR-31-5p mimics group,the miR-31-5p inhibitor+sh-HMGA1 group,and the circ_0084043+sh-HMGA1 group increased(P<0

关 键 词：circ_0084043 miR-31-5p 高迁移率族蛋白A1 动脉粥样硬化 血管平滑肌细胞 

分 类 号：R73[医药卫生—肿瘤]