姜黄素通过激活Smad1/5-Smad4-β-catenin信号通路调控孤核受体4A1在易损斑块血管新生中的作用  

Curcumin regulates the role of NR4A1 in angiogenesis of vulnerable plaques by activating the Smad1/5-Smad4-β-catenin signaling pathway

作　　者：张永霞[1] 陈志红[1] 邹润凤 ZHANG Yong-xia;CHEN Zhi-hong;ZOU Run-feng(Department of Cardiovascular Medicine,the Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,Guangdong,China)

机构地区：[1]广州医科大学附属第三医院心血管内科,广东广州510150

出　　处：《川北医学院学报》2024年第12期1598-1603,共6页Journal of North Sichuan Medical College

基　　金：广东省中药局科研项目(20222128)。

摘　　要：目的:探究姜黄素激活Smad1/5-Smad4-β-catenin信号通路调控孤核受体4A1(NR4A1)在易损斑块血管新生中的作用。方法:体外培养人脐静脉内皮细胞(HUVEC),将HUVEC分为模型组、Si-NR4A1组(NR4A1-/-HUVEC组)、姜黄素组(给予姜黄素20μg/mL干预处理),每组各10孔。在TNF-ɑ或氧化低密度脂蛋白(oxLDL)+BMP-9的刺激下行内皮细胞增殖、迁移、Matrigel基质胶管腔形成实验,并采用Matrigel基质胶法检测细胞管腔形成能力,用ELISA法检测细胞上清液炎症因子,用Transwell法检测各组迁移能力,使用Western blot法检测Smad1/5、Smad4、β-catenin的表达水平。结果:RT-qPCR结果显示,各组NR4A1表达水平比较:姜黄素组<Si-NR4A1组<模型组(P<0.05);各组细胞迁移率、细胞迁移面积、细胞管腔形成能力、细胞增殖能力比较:姜黄素组<Si-NR4A1组<模型组(P<0.05);与Si-NR4A1组及模型组相比,姜黄素组肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、内皮素1(ET-1)、白细胞介素12(IL-12)水平下降(P<0.05);各组细胞的Smad1、Smad5、Smad4的蛋白表达水平比较:姜黄素组>Si-NR4A1组>模型组(P<0.05)。各组细胞的β-catenin蛋白表达水平比较:姜黄素组<Si-NR4A1组<模型组(P<0.05)。结论:姜黄素具有抑制细胞迁移与增殖等作用,并对内皮细胞功能起保护作用,其可能与通过NR4A1激活Smad1/5-Smad4-β-catenin表达有关。Objective:To explore the role of curcumin in activating Smad homologous protein 1/5(Smad1/5)-(Smad homologous 4(Smad4)protein)-β-catenin signaling pathway in regulating the angiogenesis of the orphan nucleus receptor 4A1(NR4A1)in vulnerable plaques.Methods:Human umbilical vein endothelial cells(HUVEC)were cultured in vitro,and HUVEC was divided into model group(HUVE group),Si-NR4A1 group(NR4A1-/-HUVEC group)and curcumin group(treated with 20μg·mL-1 curcumin intervention)with 10 Wells in each group.The TNF-ɑor oxygenic low-density lipoprotein(oxLDL)+BMP-9 stimulation induced endothelial cell proliferation,migration and Matrigel matrix rubber tube lumen formation experiments were performed.Matrigel matrix glue method was used to detect the lumen formation ability of cells,and the inflammatory factors of cell supernatant were detected by ELISA.Transwell method was used to detect the migration ability of each group,and Western Blot method was used to detect the expression levels of Smad1/5,Smad4 andβ-catenin.Results:RT-qPCR results showed that expression of NR4A1 in Si-NR4A1 and curcumin groups decreased compared with model group(P<0.05),and NR4A1 in curcumin group was lower than Si-NR4A1 group(P<0.05).Compared with model group,the cell mobility,cell migration area,cell lumen formation capacity and cell proliferation capacity of curcumin group and Si-NR4A1 group decreased(P<0.05),and the curcumin group was lower than the Si-NR4A1 group(P<0.05).However,compared with Si-NR4A1 group,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),endothelin-1(ET-1),and interleukin-12(IL-12)levels in curcumin group were decreased(P<0.05).Compared with model group,protein expressions of Smad1,Smad5 and Smad4 from Si-NR4A1 group and curcumin group were increased(P<0.05),and curcumin group was higher than Si-NR4A1 group(P<0.05).β-catenin protein expression in Si-NR4A1 group and curcumin group was lower than that of model group(P<0.05),curcumin group was lower than Si-NR4A1 group(P<0.05).Conclusion:Curcumin can inhibit cell migrat

关 键 词：姜黄素 孤核受体4A1 Smad同源物1/5-Smad同源物4-β-连环蛋白 易损斑块 血管新生 

分 类 号：R543.3[医药卫生—心血管疾病]