胡彦武;李海涛;刘凯;闫梦彤;张洋;任立群;
HU Yan-wu;LI Hai-tao;LIU-Kai;YAN Meng-tong;ZHANG Yang;REN Li-qun;School of Pharmaceutics and Food Science,Tonghua Normal University;Affiliated Hospital of Changchun University of Traditional Chinese Medicine;School of Pharmacy,Jilin University;

• 1:通化师范学院制药与食品科学学院
• 2:长春中医药大学附属医院
• 3:吉林大学药学院

摘要(Abstract)：

研究淫羊藿苷(ICA)对氧化低密度脂蛋白(ox-LDL)诱导血管平滑肌细胞(VSMC)增殖的抑制作用,以及对增殖细胞核抗原(PCNA)表达和丝裂原活化蛋白激酶(MAPK)信号通路的影响。以ox-LDL(50 mg·L-1)诱导VSMC增殖,采用MTT法检测ICA对ox-LDL诱导VSMC增殖的抑制作用;Real-time PCR和Western blot方法检测ICA对ox-LDL诱导增殖VSMC内细胞周期相关蛋白PCNA基因及蛋白表达的影响;Western blot方法检测ICA对ox-LDL诱导增殖VSMC内p38MAPK,ERK1/2,JNK信号通路相关信号蛋白磷酸化的影响。结果显示,经ox-LDL(50 mg·L-1)刺激后,VSMC增殖活性显著增强,细胞存活率显著提升,S期、G2/M期细胞显著增加,G0/G1期细胞显著减少;同时,PCNA基因及蛋白表达水平显著提升;不同浓度的ICA(40,20,10μmol·L-1)可显著抑制ox-LDL诱导的VSMC增殖,减少S期、G2/M期细胞,增加G0/G1期细胞,降低ox-LDL诱导的PCNA基因及蛋白的表达水平,下调ox-LDL诱导的p-p38和p-ERK1/2蛋白的表达,但不影响p-JNK蛋白的表达。上述结果表明,ICA对ox-LDL诱导的VSMC增殖具有抑制作用,可能通过降低PCNA表达和阻断p38MAPK和ERK1/2信号通路抑制VSMC增殖。
This paper was aimed to study the effects of icariin( ICA) on the proliferation of vascular smooth muscle cell( VSMC)induced by oxidized low density lipoprotein( ox-LDL),and the molecular mechanism of the expression of proliferating cell nuclear antigen( PCNA) and MAPK signaling pathway. In this study,VSMC was induced by ox-LDL( 50 mg·L-1),the effect of ICA on the proliferation of VSMC was detected by MTT assay,Western blot and Real-time PCR. The results showed that after stimulation of ox-LDL,the proliferation activity of VSMC was increased,S phase,G2/M phase cells were increased,G0/G1 phase cells were decreased,PCNA protein expression was enhanced; ICA( 40,20,10 μmol·L-1) could effectively inhibit ox-LDL-induced VSMC proliferation,S phase and G2/M phase cells were decreased,the percentage of cells in G0/G1 phase were increased,PCNA expression was decreased,p38 MAPK and ERK1/2 activation were inhibited. These results indicate that ICA can inhibit the proliferation of VSMC by reducing the expression of PCNA and blocking the p38 MAPK and ERK1/2 signaling pathway.

关键词(KeyWords)： 淫羊藿苷;血管平滑肌细胞;增殖细胞核抗原;MAPK信号通路;细胞周期
icariin;vascular smooth muscle cell;proliferating cell nuclear antigen;MAPK signal pathway;cell cycle

Abstract:

Keywords:

基金项目(Foundation): 吉林省自然科学基金项目(20150101221JC);; 吉林省教育厅“十三五”科学技术项目(吉教科合字[2016]第247号);; 通化师范学院应用方向研究项目(2014096)

作者(Authors): 胡彦武;李海涛;刘凯;闫梦彤;张洋;任立群;
HU Yan-wu;LI Hai-tao;LIU-Kai;YAN Meng-tong;ZHANG Yang;REN Li-qun;School of Pharmaceutics and Food Science,Tonghua Normal University;Affiliated Hospital of Changchun University of Traditional Chinese Medicine;School of Pharmacy,Jilin University;

参考文献(References)：

• [1]Lacolley P,Regnault V,Nicoletti A,et al.The vascular smooth muscle cell in arterial pathology:a cell that can take on multiple roles[J].Cardiovasc Res,2012,95(2):194.
• [2]Lovren F,Teoh H,Verma S.Obesity and atherosclerosis:mechanistic insights[J].Can J Cardiol,2015,31(2):177.
• [3]胡彦武,刘凯,闫梦彤.淫羊藿苷对大鼠糖尿病心肌缺血再灌注损伤模型的治疗作用及机制研究[J].中国中药杂志,2015,40(21):4234.
• [4]Hu Yanwu,Liu Kai,Yan Mengtong,et al.Effects and mechanisms of icariin on atherosclerosis[J].Int J Clin Exp Med,2015,8(3):3585.
• [5]Huang G L,Wei Z,Ren H Y,et al.Retinoid X receptorαenhances human cholangiocarcinoma growth through simultaneous activation of Wnt/β-catenin and nuclear factor-κB pathways[J].Cancer Sci,2015,106(11):1515.
• [6]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method[J].Methods,2001,25(4):402.
• [7]Rudijanto A.The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis[J].Acta Med Indones,2007,39(2):86.
• [8]赵京山,方明星,郭浅妤,等.羟基红花黄色素A通过影响PCNA表达和MEK-ERK1/2信号通路抑制大鼠血管平滑肌细胞增殖[J].中国药理学通报,2015,31(7):984.
• [9]曹浪,邓常清.补阳还五汤苷类有效组分及其有效成分配伍对血管平滑肌细胞增殖及相关信号转导通路的影响[J].中国中药杂志,2016,41(10):1889.
• [10]Kwon H,Lee J J,Lee J H,et al.Cinnamon and its components suppress vascular smooth muscle cell proliferation by up-regulating cyclin-dependent kinase inhibitors[J].Am J Chin Med,2015,43(4):621.
• [11]Hu Yanwu,Liu Kai,Yan Mengtong,et al.Icariin inhibits oxidized low-density lipoprotein-induced proliferation of vascular smooth muscle cells by suppressing activation of extracellular signal-regulated kinase 1/2 and expression of proliferating cell nuclear antigen[J].Mol Med Rep,2016,13(3):2899.
• [12]Hu Yanwu,Sun Bo,Liu Kai,et al.Icariin attenuates high-cholesterol diet induced atherosclerosis in rats by inhibition of inflammatory response and p38 MAPK signaling pathway[J].Inflammation,2016,38(1):228.
• [13]Wang G F,Shi C G,Sun M Z,et al.Tetramethylpyrazine attenuates atherosclerosis development and protects endothelial cells from ox-LDL[J].Cardiovasc Drugs Ther,2013,27(3):199.
• [14]Yu W,Ying H,Tong F,et al.Protective effect of the silkworm protein 30Kc6 on human vascular endothelial cells damaged by oxidized low density lipoprotein(ox-LDL)[J].PLo S ONE,2013,8(6):e68746.