期刊文献+

烟酸对p38丝裂原活化蛋白激酶信号通路介导的内皮细胞功能障碍的早期干预研究

Effects of niacin on cell adhesion and early atherogenesis:involvement of the p38 mitogen-activated protein kinases pathway
原文传递
导出
摘要 目的 探讨烟酸对p38MAPK通路介导的内皮细胞功能障碍的早期干预作用及可能机制.方法 人脐静脉内皮细胞株(HUVECs),用Medium200培养基培养,实验分组:①阴性对照组:培养基;②溶血磷脂酰胆碱(LPC)不同作用时间组:培养基中加入终浓度为20 μmol/L的LPC,分别培养10 min、8h、24 h;③LPC+ p38MAPK的抑制剂(SB203580)组:培养基中加入SB203580 10 μmol/L培养1h,再分别加入LPC培养10 min、8h、24 h;④LPC+不同剂量烟酸组:培养基中分别加入终浓度为0.25、0.5、1 mmol/L烟酸培养18h,再加入LPC培养10 min、8h及24 h.应用Western blot定量分析检测内皮细胞磷酸化的p38 MAPK(pp38 MAPK)、细胞间黏附分子(ICAM-1)蛋白含量,实时定量PCR方法检测内皮细胞ICAM-1 mRNA表达,细胞免疫荧光方法检测LPC诱导的ICAM-1蛋白表达.结果 ICAM-1的蛋白表达LPC 24 h组为0.786±0.021,LPC+烟酸(1 mmol/L)组培养24 h为0.487±0.015,LPC+ SB203580组培养24 h为0.461±0.011,LPC+烟酸组和LPC+ SB203580组均低于LPC 24 h组,差异有统计学意义(F=6.3,P<0.01),但均未达到阴性对照组水平(0.134±0.012).pp38MAPK蛋白在LPC 10 min组最高,为0.47±0.02,烟酸能降低pp38MAPK,LPC+烟酸(1 mmol/L)组为0.07±0.02,LPC+ SB203580为0.11±0.02,均低于LPC 10 min组(F=91.91,P<0.01).加入LPC培养8h后,ICAM-1 mRNA的表达(8.16±0.15),高于阴性对照组(1.00±0.02),差异有统计学意义(t=24.34,P<0.01);与LPC培养8h比较,烟酸降低LPC诱导的ICAM-1 mRNA的表达,LPC+烟酸(1 mmol/L)组为3.85±0.14(F =8.06,P<0.01),而SB203580则不能有效的降低ICAM-1的mRNA的表达(8.09±0.11).结论 在LPC诱导的HUVECs中,ICAM-1的蛋白与mRNA的表达均明显增强,pp38MAPK蛋白明显增强,烟酸干预可降低ICAM-1的蛋白与mRNA的表达,同时亦可降低pp38 MAPK的蛋白表达,p38MAPK的抑制剂SB203580仅能降低ICAM-1的蛋白的表达,而不能影响其mRNA表达,其作用机制有待进一步研究. Objective To examine the effects of niacin on lysophosphatidylcholine (LPC)-induced intercellular adhesion molecule-1 (ICAM-1), and gained insight to the mechanisms. Method Human umbilical vein endothelial cell line was cultured using Medium 200 medium in incubator at 37 ℃ and 5% CO2 condition. Experimental groups: (1) the negative control group: medium; (2) LPC different time groups:the medium added with 20 μmol/L final concentration of LPC, were cultured for 10 min and 8 h, 24 h ; ( 3 ) LPC ± p38-mitogen-activated protein kinase (p38MAPK) inhibitor ( SB203580 ) group: the medium added with 10 txmol/L p38 MAPK inhibitor (SB203580) was cultured for 1 h, then human umbilical vein endothelial cells (HUVECs) added with the LPC were cultured for 10 rain, 8 h and 24 h. (4) LPC ± different niacin dose group ~ after separately adding with O. 25, O. 5, 1 mmol/L niacin, the cells were cultured for 18 h, then HUVECs added with the LPC were cultured for 10 min, 8 h and 24 h. Cell concentration in each group was 5× 105/ml, inoculated in 6-well plates, each well 1 ml. Detected by Western blot analysis of pp38MAPK, ICAM-1 protein content, real-time quantitative PCR to detect endothelial cell ICAM-! mRNA expression, cell immunofluorescence to detect LPC-induced ICAM-1 protein expression. Result In LPC 24 h group, the expression of ICAM-1 protein was significantly increased 0. 786 ± 0.02, the LPC ± niacin group, ICAM-1 protein levels (0. 487 ±0. 015) was significantly lower than the LPC 24 h group (P 〈 0. 01 ), in LPC ± SB203580 intervention group, ICAM-1 protein levels (0. 461 ± 0.011 ) was significantly lower than that of the LPC 24 h group ( P 〈 0.01 ), but did not reach the level of the control group. Adding LPC to culture for 10 min, phosphorylation of p38MAPK (pp38MAPK) reached its peak (0. 47 ± 0. 02), niacin could reduce the pp38MAPK (0.07 -± 0.02), SB203580 could also reduce its activity (0. 11 ± 0.02 ) ~ Adding LPC to culture for 8 h, ICAM-1 mRNA expression (8. 16 ±-0. 15) compared with the control group ( 1.00 _± 0.02 ) had a significant increase ( t = 24. 34, P 〈 0. 01 ). Compared with the LPC 8 h, niacin reduced LPC-induced ICAM-1 mRNA expression ( 3.85 ± 0. 14 ) , and showed a dose-dependent manner (F = 8. 06, P 〈 0. 01 ), while SB203580 could not effectively reduce the ICAM-1 mRNA (8. 09 ± 0. 11 ). Conclusion Niacin prevented LPC-induced endothelial dysfunction by reducing expression of ICAM-1. These mechanisms appeared to be at least partly mediated by suppression of the pp38MAPK in endothelial ceils. These pleiotropic effects of niacin may potentially contribute to the beneficial effects of risk reduction for atherosclerotic disease.
出处 《中华儿科杂志》 CAS CSCD 北大核心 2013年第11期825-830,共6页 Chinese Journal of Pediatrics
基金 山东省优秀中青年科学家科研奖励基金(BS2010YY056)
关键词 内皮细胞 P38丝裂原活化蛋白激酶类 烟酸类 Endothelial cell p38 mitogen-activated protein kinases Nicotinic acids
  • 相关文献

参考文献18

  • 1Schwartz CJ,VaIente AJ,Sprague EA.A modem view ofatherogenesis.Am J Cardiol,1993,71:9B-14B.
  • 2Principi M,Scicchitano P,Scicchitano P,et al.Endothelialfunction and cardiovascular risk in active inflammatory boweldiseases.J Crohns Colitis,2013,doi:pii:SI873-9946(13)00065-2.10.1016/j.crohns.2013.02.001.Epub ahead ofprint.
  • 3McGinn S,Ssad P,Pomnnik,et al.High gouclose-mediatedeffects on endothelial cell proliferation occur via p38MAP kinase.Am J Physiol Endocrinol Metab,2003,285 ;E708-E717.
  • 4Christopher S,Ramadan A,Sameer H,et al.Revisiting niacin:reviewing the evidence.J Clin Lipidol,2007,I:248-255.
  • 5Aoyama T,Fujiwara H,Masaki T,et al.Induction of lectin-likeoxidized LDL receptor by oxidized LDL andlysophosphatidylcholine in cultured endothelial cells.J Mol CellCardiol,1999,31:2101-2114.
  • 6Tavintharan S,Lim SC,Sum CF.Effects of niacin on celladhesion and early atherogenesis:biochemical and functionalfindings in endothelial cells.Basic Clin Pharmacol Toxicol,2009,104:206-210.
  • 7Erdogan A,Schaefer MB,Kuhlmann CR,et al.Activation of Ca2+-activated potassium channels is involved iniysophosphatidylcholine-induced monocyte adhesion to endothelialcells.Atherosclerosis,2007,190:100-105.
  • 8Yamakawa T,Ohnaka K,Tanaka S,et al.Cyclooxygenase-2induction by lysophosphatidylcholine in cultured rat vascularsmooth muscle cells:involvement of p38MAPK pathway.BiomedRes,2008,29:1-8.
  • 9Shen J,DiCorleto PE.ADP stimulates human endothelial cellmigration via P2Y1 nucleotide receptor-mediated mitogen-activatedprotein kinase pathways.Circ Res,2008,102:448456.
  • 10Ganji SH,Qin S,Zhang L,et al.Niacin inhibits vascularoxidative stress,redox-sensitive genes,and monocyte adhesion tohuman aortic endothelial cells.Atherosclerosis,2009,202:68.

二级参考文献19

  • 1于永慧,董波,汪翼.LOX-1在动脉粥样硬化中的作用[J].国外医学(心血管疾病分册),2004,31(5):280-283. 被引量:6
  • 2黄东,葛均波,梁春,罗育坤,贾庆哲,王克强.凝集素样氧化低密度脂蛋白受体在兔自体静脉移植物粥样硬化病变中的表达及洛沙坦干预作用的研究[J].中华心血管病杂志,2004,32(10):915-918. 被引量:12
  • 3朱敏恒,吴越,陈虹,田卉,刘洪海.烟酰水杨酸对Collagen、ADP、AA诱导的兔血小板聚集的影响[J].中国药理学通报,2005,21(10):1232-1234. 被引量:9
  • 4O'Gorman CS,O'Neill MB,Conwell LS. Considering statins for cholesterol-reduction in children if lifestyle and diet changes do not improve their health:A review of the risks and benefits[J].Vasc Health Risk Manag,2010,(07):1-14.
  • 5McKenney J,Bays H,Koren M. Safety of extended-release niacin/laropiprant in patients with dyshpidemia[J].J Clin Lipidol,2010,(02):105-112.doi:10.1016/j.jacl.2010.02.002.
  • 6Rubenfire M,Brook RD,Rosenson RS. Treating mixed hyperlipidemia and the atherogenic lipid phenotype for prevention of cardiovascular events[J].American Journal of Medicine,2010,(10):892-898.doi:10.1016/j.amjmed.2010.03.024.
  • 7Olsson AG. Recent advances in preventing cardiovascular disorders by managing lipid levels[J].F1000 Med Rep,2010.66-80.
  • 8AIM-HIGH Investigators. The role of niacin in raising high-density lipoprotein cholesterol to reduce cardiovascular events in patients with atherosclerotic cardiovascular disease and optimally treated low-density lipoprotein cholesterol Rationale and study design.The Atherothrombosis Intervention in Metabolic syndrome with low HDL/high triglycerides:Impact on Global Health outcomes (AIM-HIGH)[J].American Heart Journal,2011,(03):471-477.
  • 9施新猷.医用实验动物学[M]北京:人民军医出版社,1999388.
  • 10华琦.单纯肥胖症基础与临床[M]北京:北京科学技术出版社,199858.

共引文献9

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部