冠心合剂对冠心病PCI术后患者血管内皮功能的影响及其主要成分对TNF-α损伤人脐静脉内皮细胞保护作用机制的研究
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摘要
目的观察冠心合剂对冠心病患者PCI术前后血清中NO、ET-1水平的影响以及探讨冠心合剂主要成分(黄芪甲苷、槲皮素、异鼠李素、β谷甾醇)对TNF-α损伤人脐静脉内皮细胞(HUVECs)保护作用的机制。
方法临床观察选取新近冠脉造影阳性且植入支架的冠心病患者36例,随机分为对照组和治疗组,分别给予基础治疗和基础治疗加冠心合剂治疗,疗程二周后,Elisa法测定两组血清中NO、ET-1水平。细胞实验采用传代后的人脐静脉内皮细胞培养作为实验对象,随机分为正常对照组、模型组、黄芪甲苷组、槲皮素组、异鼠李素组、β谷甾醇组。据前期试验结果,除正常组和模型组外,药物组以各药物成分(浓度分别为黄芪甲苷80μg/ml、槲皮素30μg/ml、异鼠李素20μg/ml、β-谷甾醇20μg/ml)与HUVECs共培养18小时后,模型组和各药物组分别再加入浓度为200ng/ml的TNF-α刺激HUVECs6小时后,分别用RT-PCR检测细胞黏附分子-1、血管细胞粘附分子-1、E-选择素、PAI-1mRNA的表达；以及流式细胞术和Western-blot检测激活蛋白AP-1、核因子NF-кB的表达水平以及HUVECs细胞的凋亡率。
结果临床观察结果与对照组相比,治疗组NO水平升高有统计学意义(P<0.05),ET-1水平降低明显(P<0.05),和冠心合剂治疗前NO水平亦明显升高(P<0.05),ET-1水平降低明显(P<0.05)；两组患者术后与术前相比,NO和ET-1水平两组间差异均无统计学意义(P>0.05)。细胞实验发现①与正常组比较,模型组ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达有极显著性差异,具有统计学意义(P<0.01)；与模型组相较,黄芪甲苷、槲皮素、异鼠李素均可降低TNF-α诱导的HUVECs的ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达(P<0.05、P<0.05、P<0.05),而与正常组相较差异无统计学意义(P>0.05、P>0.05、P>0.05)；而p-谷甾醇组对ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达抑制不明显,与模型组相较无统计学意义(P>0.05)。②流式细胞术检测细胞凋亡,与正常组比较,模型组凋亡率有显著统计学意义(P<0.01)；黄芪甲苷、槲皮素、异鼠李素均可明显降低细胞凋亡率,与模型组相较,黄芪甲苷组具有极显著统计学意义(P<0.01),槲皮素组和异鼠李素组差异也具有显著统计学意义(P<0.05、P<0.05),其中黄芪甲苷组较槲皮素组、异鼠李素组有统计学意义(P<0.05),但与正常组相较,槲皮素组、异鼠李素组仍有明显差异(P<0.05)；而p-谷甾醇组对凋亡的抑制作用不明显,差异无统计学意义(P>0.05)。③与正常组比较,模型组激活蛋白AP-1、核因子NF-кB的表达显著上调,有显著统计学意义(P<0.01)：黄芪甲苷、槲皮素、异鼠李素均可降低TNF-α诱导的HUVECs的激活蛋白AP-1、转录因子NF-кB的表达(P<0.01、P<0.01、P<0.01),p-谷甾醇组对AP-1和NF-кB的表达无明显抑制作用,差异无统计学意义(P>0.05)。
结论冠心合剂能够升高冠心病患者PCI术后血清NO并降低ET-1水平。而其主要成分黄芪甲苷等对TNF-α损伤人血管内皮细胞的保护作用可能源于通过对激活蛋白AP-1、核因子NF-кb蛋白表达的抑制,减少细胞凋亡的同时,调节ICAM-1、VCAM-1、E.sel、PAI-1mRNA的表达等途径而实现的。
Objective To observe the influences on ET-1and NO in plasma with the treatment of Guanxin decoction on the basic therapy to the CHD patients treated by PCI in clinical. And in cell experimental reseacrh,we will investigate part protective mechanism of Guanxin Decoction on the HUVECs exposed to TNF-a. Accomplishment of the topic will provide some important experimental evidences for treatment mechanisms of Guanxin Decoction
Method In clinical Thirty-six CHD patients were randomly divided into the trial group and the control group after treat with PCI recently. The patients of trial group were treated by the Guanxin decoction, and simultaneously accept conventional therapy. as same as the control group. The period of treatment was two week. The level of ET-1and NO in plasma of two groups after treatment were compared by Elisa.In cell experiment, The cultured HUVECs from third to sixth generations were randomly divided into the trial group and the control group(model group, control group, Astragaloside group, Quercetin group, Isorhamnetin group, β-Sitosterol group).Except model group and control group, every research groups cultured with different main active ingredient of Guanxin Decoction level (Astragaloside80μg/ml,Quercetin30μg/ml,Isorhamnetin20ug/ml,β-Sitosterol20μg/ml)for18hours,And then were added TNF-a (2ng/ml),after6hours,ICAM-1、VCAM-1,E-select,PAI-1were detected by reverse transcriptase polymerase chain reaction (RT-PCR) in HUVECs; AP-1and NF-Kb were detected by ANNEXIN-V and Western-blot.
Result After treatment in clinical, the level of ET-1in plasma of the trial group reduced and the level of NO in plasma rose. Compared with that before treatment and the control group, the results had significant difference (P<0.05),and the level of ET-1and NO in plasma of two groups was no significant difference compared with before PCI (P>0.05). In cell experimental reseacrh,we found①After cultured by TNF-a, Comparing with normal group, the levels of ICAM-1,VCAM-1,E-select,PAI-1mRNA of model group changed insignificantly (P<0.01); except β-Sitosterol group,there were no statistical differences in levels of ICAM-1,VCAM-1, E-select and PAI-1mRNA between treated group and normal control group (P>0.05).And comparing with model group,the expression of ICAM-1,,VCAM-1,E-select and PAI-1mRNA of Astragaloside group,Quercetin group and Isorhamnetin group changed significantly (P<0.01, P<0.01,P<0.01)but β-Sitosterol group.②Comparing with normal group,apoptosis percentage of HUVECs exposed to TNF-a are higher than those in normal group(P<0.01), exsept β-Sitosterol group,there were no statistical differences between treated group and normal control group (P>0.05); And comparing with model group, apoptosis percentage of HUVECs decreased significantly (P<0.01,P<0.01,P<0.01) but P-Sitosterol group.③Comparing with normal group, expression of AP-1and NF-Kb of model group exposed to TNF-a are higher than those in normal group(P<0.01),there were no statistical differences between treated group and normal control group (P>0.05); And comparing with model group, expression of AP-1and NF-Kb were inhibited significantly (P<0.01, P<0.01, P<0.01)but β-Sitosterol group(P>0.05).
Conclusion Guanxin decoction can improve the level of NO in plasma of CHD patients,and reduce the level of ET-1in clinical. And in cell experimental reseacrh the main active ingredient of Guanxin decoction(Astragaloside,Quercetin and Isorhamnetin) can decrease the expression of ICAM-1,VCAM-1,E-select and PAI-1mRNA on HUVECs exposed to TNF-a;at the same time,can inhibite the expression of AP-1and NF-Kb;and degree apoptosis percentage of HUVECs,but the β-Sitosterol have none of the above.So anti-atherosclerosis of Guanxin decoction maybe realize in this way.
方法临床观察选取新近冠脉造影阳性且植入支架的冠心病患者36例,随机分为对照组和治疗组,分别给予基础治疗和基础治疗加冠心合剂治疗,疗程二周后,Elisa法测定两组血清中NO、ET-1水平。细胞实验采用传代后的人脐静脉内皮细胞培养作为实验对象,随机分为正常对照组、模型组、黄芪甲苷组、槲皮素组、异鼠李素组、β谷甾醇组。据前期试验结果,除正常组和模型组外,药物组以各药物成分(浓度分别为黄芪甲苷80μg/ml、槲皮素30μg/ml、异鼠李素20μg/ml、β-谷甾醇20μg/ml)与HUVECs共培养18小时后,模型组和各药物组分别再加入浓度为200ng/ml的TNF-α刺激HUVECs6小时后,分别用RT-PCR检测细胞黏附分子-1、血管细胞粘附分子-1、E-选择素、PAI-1mRNA的表达；以及流式细胞术和Western-blot检测激活蛋白AP-1、核因子NF-кB的表达水平以及HUVECs细胞的凋亡率。
结果临床观察结果与对照组相比,治疗组NO水平升高有统计学意义(P<0.05),ET-1水平降低明显(P<0.05),和冠心合剂治疗前NO水平亦明显升高(P<0.05),ET-1水平降低明显(P<0.05)；两组患者术后与术前相比,NO和ET-1水平两组间差异均无统计学意义(P>0.05)。细胞实验发现①与正常组比较,模型组ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达有极显著性差异,具有统计学意义(P<0.01)；与模型组相较,黄芪甲苷、槲皮素、异鼠李素均可降低TNF-α诱导的HUVECs的ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达(P<0.05、P<0.05、P<0.05),而与正常组相较差异无统计学意义(P>0.05、P>0.05、P>0.05)；而p-谷甾醇组对ICAM-1、VCAM-1、E-sel、PAI-1mRNA的表达抑制不明显,与模型组相较无统计学意义(P>0.05)。②流式细胞术检测细胞凋亡,与正常组比较,模型组凋亡率有显著统计学意义(P<0.01)；黄芪甲苷、槲皮素、异鼠李素均可明显降低细胞凋亡率,与模型组相较,黄芪甲苷组具有极显著统计学意义(P<0.01),槲皮素组和异鼠李素组差异也具有显著统计学意义(P<0.05、P<0.05),其中黄芪甲苷组较槲皮素组、异鼠李素组有统计学意义(P<0.05),但与正常组相较,槲皮素组、异鼠李素组仍有明显差异(P<0.05)；而p-谷甾醇组对凋亡的抑制作用不明显,差异无统计学意义(P>0.05)。③与正常组比较,模型组激活蛋白AP-1、核因子NF-кB的表达显著上调,有显著统计学意义(P<0.01)：黄芪甲苷、槲皮素、异鼠李素均可降低TNF-α诱导的HUVECs的激活蛋白AP-1、转录因子NF-кB的表达(P<0.01、P<0.01、P<0.01),p-谷甾醇组对AP-1和NF-кB的表达无明显抑制作用,差异无统计学意义(P>0.05)。
结论冠心合剂能够升高冠心病患者PCI术后血清NO并降低ET-1水平。而其主要成分黄芪甲苷等对TNF-α损伤人血管内皮细胞的保护作用可能源于通过对激活蛋白AP-1、核因子NF-кb蛋白表达的抑制,减少细胞凋亡的同时,调节ICAM-1、VCAM-1、E.sel、PAI-1mRNA的表达等途径而实现的。
Objective To observe the influences on ET-1and NO in plasma with the treatment of Guanxin decoction on the basic therapy to the CHD patients treated by PCI in clinical. And in cell experimental reseacrh,we will investigate part protective mechanism of Guanxin Decoction on the HUVECs exposed to TNF-a. Accomplishment of the topic will provide some important experimental evidences for treatment mechanisms of Guanxin Decoction
Method In clinical Thirty-six CHD patients were randomly divided into the trial group and the control group after treat with PCI recently. The patients of trial group were treated by the Guanxin decoction, and simultaneously accept conventional therapy. as same as the control group. The period of treatment was two week. The level of ET-1and NO in plasma of two groups after treatment were compared by Elisa.In cell experiment, The cultured HUVECs from third to sixth generations were randomly divided into the trial group and the control group(model group, control group, Astragaloside group, Quercetin group, Isorhamnetin group, β-Sitosterol group).Except model group and control group, every research groups cultured with different main active ingredient of Guanxin Decoction level (Astragaloside80μg/ml,Quercetin30μg/ml,Isorhamnetin20ug/ml,β-Sitosterol20μg/ml)for18hours,And then were added TNF-a (2ng/ml),after6hours,ICAM-1、VCAM-1,E-select,PAI-1were detected by reverse transcriptase polymerase chain reaction (RT-PCR) in HUVECs; AP-1and NF-Kb were detected by ANNEXIN-V and Western-blot.
Result After treatment in clinical, the level of ET-1in plasma of the trial group reduced and the level of NO in plasma rose. Compared with that before treatment and the control group, the results had significant difference (P<0.05),and the level of ET-1and NO in plasma of two groups was no significant difference compared with before PCI (P>0.05). In cell experimental reseacrh,we found①After cultured by TNF-a, Comparing with normal group, the levels of ICAM-1,VCAM-1,E-select,PAI-1mRNA of model group changed insignificantly (P<0.01); except β-Sitosterol group,there were no statistical differences in levels of ICAM-1,VCAM-1, E-select and PAI-1mRNA between treated group and normal control group (P>0.05).And comparing with model group,the expression of ICAM-1,,VCAM-1,E-select and PAI-1mRNA of Astragaloside group,Quercetin group and Isorhamnetin group changed significantly (P<0.01, P<0.01,P<0.01)but β-Sitosterol group.②Comparing with normal group,apoptosis percentage of HUVECs exposed to TNF-a are higher than those in normal group(P<0.01), exsept β-Sitosterol group,there were no statistical differences between treated group and normal control group (P>0.05); And comparing with model group, apoptosis percentage of HUVECs decreased significantly (P<0.01,P<0.01,P<0.01) but P-Sitosterol group.③Comparing with normal group, expression of AP-1and NF-Kb of model group exposed to TNF-a are higher than those in normal group(P<0.01),there were no statistical differences between treated group and normal control group (P>0.05); And comparing with model group, expression of AP-1and NF-Kb were inhibited significantly (P<0.01, P<0.01, P<0.01)but β-Sitosterol group(P>0.05).
Conclusion Guanxin decoction can improve the level of NO in plasma of CHD patients,and reduce the level of ET-1in clinical. And in cell experimental reseacrh the main active ingredient of Guanxin decoction(Astragaloside,Quercetin and Isorhamnetin) can decrease the expression of ICAM-1,VCAM-1,E-select and PAI-1mRNA on HUVECs exposed to TNF-a;at the same time,can inhibite the expression of AP-1and NF-Kb;and degree apoptosis percentage of HUVECs,but the β-Sitosterol have none of the above.So anti-atherosclerosis of Guanxin decoction maybe realize in this way.
引文
1.叶任高,陆再英.内科学.北京：人民卫生出版社,2004.272
2.陈纪林.防治动脉粥样硬化的新动向[J].中国循环杂志,2001,16(3)：163-166
3.王利民.黏附分子和动脉粥样硬化[J].心血管病学进展,2005,26(5)：494-498
4. Badia E, Sacanella E, Fernandez-Sola J, et al. Decreased tumor necrosis factor, induced adhesion of human monocytes to endothelial cells after moderate alcohol consumption[J]. Am J Clin Nutr,2004,80(1):225-230
5. Tang V, Dhirapong A, Yabes AP, et al. TNF·alpha·media ted apoptosis in vascular smooth muscle cells requires p73[J]. Am J Physiol Cell Physiol,2005,289(1):C199.206
6. Chen YG, Xu F, Zhang Y, et al. Efect ofaspirin plus clopidogrel on inflammatory markers in patients with non-ST segment elevation acute coronary syndrome[J]. Chin Med J(Engl), 2006,119(1):32-36
7. Calin Popa, Mihai G. Netea, Piet L. C. M. van Riel, Jos W. M. van der Meer, and Anton F. H. Stalenhoef.The role of TNF-in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk.J. Lipid Res.,2007; 48:751-762
8. Daniel R. Meldrum. Tumor necrosis factor in the heart. Am J Physiol Regulatory Integrative Comp Physiol,1998; 274:577
9. May Azzawi and Philip Hasleton. Tumour necrosis factor alpha and the cardiovascular system:its role in cardiac allograft rejection and heart disease Cardiovasc Res,1999; 43:850-859
10.杨富国,刘革新,董果雄,等.黄芪甲苷对缺氧/复氧损伤人脐静脉内皮细胞与中性粒细胞黏附能力及细胞核转录因子κB表达的影响[J].中国组织工程研究与临床康复,2008,12(15)：2843-2846
11.高玉霞,黄国伟,刘英华,等.大豆异黄酮对体外血管内皮细胞氧化损伤保护作用的形态学观察[J].天津医科大学学报,2003,9(1)：39-41
12.王玉英,胡涛,贾国良,等.银杏叶提取物对人脐静脉内皮细胞增殖的影响[J].第四军医大学学报,2004,25(13)：1179-1181
13.陶波,李晓宁.蒲黄对动脉粥样硬化血管内皮损伤影响的实验研究[J].中西医结合心脑血管病杂志,2004,2(4)：222-223
14.周芳,李爱媛,谢金鲜,等.蒲黄抗鹌鹑高脂血症及动脉粥样硬化的实验研究[J].中国实验方剂学杂志,2006,12(8)：48-49
15.于长青.麦胚黄酮对实验老鼠血脂水平和脂质抗氧化作用影响的研究[J].中国食品学报,2001,2(1)：30
16.祝光礼,陈铁龙,陈启兰.黄芪失笑汤为主治疗气虚血瘀型冠心病.浙江中西医结合杂志[J].2006,16(10)：625-626
17.郑筱萸.中药新药临床研究指导原则(试行)[S].北京：中国医药科技出版社,2002：68-73
18.路志正.中医诊断学[M].北京：人民卫生出版社,1995：99-101
19.祝光礼,陈铁龙,陈启兰等.黄芪失笑汤为主治疗气虚血瘀型冠心病.浙江中西医结合杂志.2006,16(10)：625-626
20.杨利.邓铁涛教授“冠心三论”[J].湖南中医药导报,2004,10(6)：8-10
21.胡丹,姜亚军.从“痰瘀热毒”论治动脉粥样硬化[J].中医药信息,2011；28(1)：5-7
22.魏自民.活血化瘀法干预动脉粥样硬化的临床研究[J].光明中医,2011；26(2)：277-278
23.王永霞,胡宇才,朱明军.冠心病心绞痛中医证候分布相关性研究[J].世界中西医结合杂志,2008,3(12)：714-716
24.郭晓燕,王宏燕等.益气活血法治疗冠心病心绞痛50例[J].中国中医急症,2003,12(3)：264
25.邵莹.益气活血法治疗冠心病心绞痛临床观察[J].现代中西医结合杂志,2006,15(4)：422-423
26.刘剑刚,徐浩,董国菊,等.血府逐瘀口服液对冠心病心绞痛患者血小板活化分子表达的影响[J].长春中医药大学学报,2007,23(1)：29-31
27.卞云云,李萍.蒙古黄芪中黄酮类成分抗超氧阴离子活性研究及构效关系分析[J].中国药学杂志,2008,43(4)：256-259
28.尚正录,尚云等.黄芪保心汤预防实验性大鼠急性心肌缺血的研究[J].上海中医药杂志,2004,38(11)：47-50
29.严辉,陈佩东,丁安伟.蒲黄的炮制及其化学成分、药理作用的研究进展[J].江苏 中医药,2005,26(1)：59-61
30.陶波,李晓宁.蒲黄对动脉粥样硬化血管内皮损伤影响的实验研究[J].中西医结合心脑血管病杂志,2004,2(4)：222-223
31.杨琦,赵岩等.五灵脂的脂溶性成分GC-MS分析[J].中国现代中药,2010；12(10)：19-21
32.李强,陆蕴如,鲁学照,等.五灵脂化学成分的研究[J].中国药学杂志,1999,34(8)：514-516
33.祝光礼,范翠娟,陈铁龙,等.黄芪失笑散对鸡胚绒毛尿囊膜促血管生成实验研究[J].中华中医药学刊,2007,25(12)：2462-2464
34.祝光礼,周凡,陈铁龙.黄芪失笑散抗心肌缺血的实验研究[J].中华中医药学刊,2008,26(1)：2232-2234
35.祝光礼,范翠娟,陈铁龙,等.黄芪失笑散对实验大鼠梗死心肌内VEGF及bFGF mRNA基因表达的影响[J].中华中医药学刊,2008,26(1)：41-43
36.祝光礼,魏丽萍,方伟,等.黄芪失笑散对血管内皮功能的影响浙江临床医学[J].2008,10(8):1016-1018
37.曹建华.辛伐他汀对冠心病患者CRP、vWF、NO及ET的影响[J].中国老年学杂志,2010,30(4)：1138-1139
38. Buchman TG,Abello PA,Smith EH,et al.Induction of heat shock response leads to apoptosis in endothelial cells previously exposed to endotoxin[J].Am J Physiol,1993,265(1 Pt 2):H165-H170
39.Vita JA, Teasure CB, Nabel EG, et al.Coronary vasomotor response to acetylcholine relates to risk factor for coronary artery disease.Circulation,1990,81:491-497
40. Carswell EA,Old LJ,Kassel RL,et al.An endotoxin-induced serum factor that causes necrosis of tumors[J].Proc Natl Acad Sci USA,1975,72(9):3666-3670
41.Dinarello CA,Marqolis NH.Cytokine-processing enzymes. Stopping the cuts[J].Curr Biol,1995,5(6):587-590
42. Strieter RM,Kunkel SL,Bone RC.Role of tumor necrosis factor-a in disease states and inflammation[J].Crit Care Med,1993,21(10 Suppl):S447-S463
43.张敬芳,王光浩.TNF-a诱导HUVEC的损伤及其意义[J].荆门职业技术学院学报, 2002,17(6)：82-84
44.宋善俊,王林林,魏文宁.TNF-a刺激的人脐静脉内皮细胞组织因子表达及其分子机制[J].中国实验血液学杂志,2003,11(2)：124-127
45.吴军,李闪,朱建华,等.TNF-a对人脐静脉内皮细胞中NO和eNOS的影响[J].浙江医学,2006,28(8)：636-684
46. Min JK, Kim YM, Kim SW, et al. TNF-related activation-induced cytokine enhances leukocyte adhesiveness:Induction of ICAM-1 and VCAM-1 via TNF receptor-associated factor and protein kinase C-dependent NF-kappaB activation in endothelial cells. J fmmunol.2005,175(1):531-540
47. Pober JS, Cotran RS.Cytokines and endothelial cell biology Physiological Reviews,1990, 70427-70451
48. Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
49.刘学俭.失笑散小考[J].四川中医,1989(11)：13-14
50. Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
51.苏敏,钟翠平.动脉粥样硬化病变中黏附分子ICAM-1、VCAM-1及E-selectin的表达[J].第三军医大学学报,2009,31(11)：1066-1068
52.饶丹.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3)：278-281
53. Bitsch A, KleneW,Muttada L,et al.A longitudinal p rospectivestudy of soluble adhesion molecules in acute stroke[J].Stroke,1998,29:2129
54.高岩,夏蕾,白明.实验性大鼠肺血栓栓塞症中ICAM-1, P-选择素的变化及当归注射液的影响[J].中国微循环,2007,11(3)：176-179
55.陈良,张梅,李长江,等.复方丹参滴丸对动脉粥样硬化粘附因子的作用[J].中国动脉硬化杂志,2007,15(2)：101-104.
56.马丽萍,秦永文,郑兴,等.黏附分子与冠状动脉病变程度的相关性分析[J].基础医学与临床,2004,24(1)：44-47.
57.任强,李自成,巫少荣,等.血管细胞黏附分子-1在动脉粥样硬化中的作用[J].临床 荟萃,2004,19(7)：419-421.
58.隋广超,胡美浩.纤溶酶原激涪物抑制因子-1蛋白分子的结构与功能.国外医学生物分子学分册.1994.16(4)：172-176
59. Schneideerman J,Sawdey MS,Keeton Mr,et al.increased type 1 plsminogen activator inhibitor gene expreasion in atherosclerotic human arteries [J]. Proc Natl Acad Sci,1992,89: 6998～7002.
60. WenzIE. Dixit VM. Ubiqutin-mediated regulation of TNFR1 signaling[J]. Cytokine Growth Factor Rev,2008,19(3):313-324
61.Christman JW,lancaster LH,Blackwell TS.Nuclear factor kappa-B:a pivotal role in the system-inflammatory response and new target for the rapy.Intensive Care Med,1998,24(11):1131-1138
62. Sergei I. Grivennikov. Michael Karin. Dangerous liaisons:STAT3 and NF-KB collab oration and crosstalk in cancer [J]. Cytok Growth,2010,21(1):11-19
63. Varfolomeev E, Goneharov T, Fedorova AV, et al. C-IAP1 and c-IAP2 are critical mediators of tumor necrosis factor-a(TNF-a)-in-duced NF-KB activation[J]. Biol Chem, 2008,283(36):242-259
64.Wang P, Qiu W, Dudgeon C,et al. PUMA is direcfly activated by NF-KB and contributes to TNF-α-induced apoptosis[J]. Cell Death Differ,2009.16(9):192-202
65. Karin M,Greten FR. NF-KB:linking inflarmnation and immunity to cancer development and progression[J]. Nat Rev Immunol,2005,5(10):749-59
66. Hess J,Angel P,Schorpp-Kistner M.AP-1 subunits:quarrel and harmony among siblings[J]. J Cell Sci,2004,117(25):5965-5973
67. TouyzRM. Deschepper C, Park JB. Inhibition of mitogen activated protein/extracellular signal-regulated kinase improves endothelial function and attenuates Ang II-induced contractility of mesenteric resistance arteries from spontaneously hypertensive rats [J]. J Hypertens.2002,20(6):1127-1134
68. Kipsbidze N. Dangas G, Tsapenko M, et al. Role of the endothelium in modulating neointimal formation:vaseoloprotective approaches to attenuate restenosis after percutaneous coronary interventions[J]. J Am Coll Cardiol,2004,44(4):733-739
69. Dragomir E, Tircol M, Manduteanu I, et al. Aspirin and PPAR-alpha activators inhibit monocyte chemoattractant protein-1 expression induced by high glucose concentration in human endothelial cells [J]. Vaseul Pharmacol,2006,44(6):440-449
1.Carswell EA,Old LJ,Kassel RL,et al.An endotoxin-induced serum factor that causes necrosis of tumors [J].Proc Natl Acad Sci USA,1975,72(9):3666-3670
2.Dinarello CA,Marqolis NH.Cytokine-processing enzymes. Stopping the cuts[J].Curr Biol,1995,5(6):587-590
3.Aderem AA.How cytokines signal messages with in cells[J].J Infect Dis,1993,167(Suppl 1):S2-S7
4.Douzinas EE,Tsidemiadou PD,Pitaridis MT,et al.The regional production of cytokines and lactate in sepsis-related multiple organ failure [J].Am J Respir Crit Care Med,1997,155(1):53-59
5.Strieter RM,Kunkel SL,Bone RC.Role of tumor necrosis factor-a in disease states and inflammation[J].Crit Care Med,1993,21(10 Suppl):S447-S463
6.王今达,脓毒症：感染性M O D S的预防[J].中国危重病急救医学,1999,11(8)：453-455.
7.陶晓根.糖皮质激素抵抗与脓毒血症[J].中国危重病急救医学,2001,13(5)：307-309
8.Buchman TQAbello PA,Smith EH,et al.Induction of heat shock response leads to apoptosis in endothelial cells previously exposed to endotoxin[J].Am J Physiol,1993,265(1 Pt 2):H165-H170
9.赵克温.人脐带静脉血管内皮细胞的体外培养及其凋亡的研究[J].山东大学学报,2002,37(3)：268-271
10.金惠铭,刘清行,张国平,等.TNFa引起的微血管内皮细胞功能障碍及其分子机制[J].中国病理生理杂志,2000,16(10)：940-941
11.董红梅,喻伦银,贾宗智,等.TNFa对培养的脐静脉内皮细胞的损伤作用[J].临床与实验病理学杂志,2002,18(4)：401-403
12.Pober JS, Cotran RS.Cytokines and endothelial cell biology Physiological Reviews,1990, 70427-70451.
13.Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
14.Sedgwick JD, Riminton DS.Cyster JG, et al.Tumor necrosis factora master-regulator of leukocyte movement.Immunol Today.2000,21 (3):110-113
15.Krown KA,Page MT,Nguyen C,et al.Tumor necrosis facor-a induced apoptsis in cardiac myocytes[J].JClin Invest,1996,98(12):2854-2865
16.周亚峰.肿瘤坏死因子对人血管内皮细胞凋亡的影响[J].苏州大学学报(医学版),2004,24(5)：620-623
17.Depraetere V,Golstein P.Fas and other cell death signaling pathways[J].Semin Immunol,1997,9(2):93-107
18.Duan H,Dixit VM.RAIDD is a new death adapt or molecule [J]. Nature,1997,385 (6611):86-89
19.Laster SM,Wood JG,Gooding LR.Tumor necrosis factor can induce both apoptosis and necrosic forms of cellysis[J] J Immunol,1988,141(8):2629-2634
20.姜华,张艳,王辰.益气活血复方含药血清对人脐静脉内皮细胞TLR4/NF-KB信号通路及TNF-a ICAM-1 mRNA表达的影响[J],辽宁中医药大学学报,2010(12)4：47
21.臧运华,唐明,郭瑞友等,溶栓颗粒含药血清对TNF-α损伤血管内皮细胞VEGF表达影响的研究[J],中国中医药科技,2011,18(1)：15-16
22.祝光礼,方小江,彭哲,等.冠心合剂对人脐静脉内皮细胞分泌血管生长因子bFGF的影响[J].浙江中西医结合杂志,2011,21(9)：596-598
23.祝光礼,魏丽萍,方伟,等.黄芪失笑散对实验大鼠主动脉和OX-LDL诱导下人脐静脉内皮细胞ICAM-1、VCAM-1 mRNA基因表达的影响[J].中华中医药学刊,2008,26(10)：2095-2098
24.尚改萍,文志斌,李俊成等,补阳还五汤对TNFa诱导血管内皮细胞释放vWF及表达组织因子的影响[J],湖南医科大学学报,2000,25(2)：129-131
25.黄建林,林灼锋,罗敏琪等,青藤碱抑制TNF-α诱导人脐静脉内皮细胞VCAM-1表达[J],中国病理生理杂志,2007,23(4)：634-638
26.叶泽兵,汪凡军,张刚庆等,黄芪注射液对TNF-α诱导人脐静脉内皮细胞炎症因子的影响及其分子机制[J],南方医科大学学报,2009,29(6)：1137-1140
27.游洋,段岩,张效林等,黄芪水提取物对TNF-α诱导的小鼠动脉内皮细胞VCAM-1表达的影响[J],中国动脉硬化杂志2011(19)3：187-190
28.刘蓓,朱海燕,高永红,黄芪多糖对TNF-α诱导心脏微血管内皮细胞黏附分子基因转录及p38MAPK信号通路的影响[J],世界中医药,2011(6)3：263-265
2.陈纪林.防治动脉粥样硬化的新动向[J].中国循环杂志,2001,16(3)：163-166
3.王利民.黏附分子和动脉粥样硬化[J].心血管病学进展,2005,26(5)：494-498
4. Badia E, Sacanella E, Fernandez-Sola J, et al. Decreased tumor necrosis factor, induced adhesion of human monocytes to endothelial cells after moderate alcohol consumption[J]. Am J Clin Nutr,2004,80(1):225-230
5. Tang V, Dhirapong A, Yabes AP, et al. TNF·alpha·media ted apoptosis in vascular smooth muscle cells requires p73[J]. Am J Physiol Cell Physiol,2005,289(1):C199.206
6. Chen YG, Xu F, Zhang Y, et al. Efect ofaspirin plus clopidogrel on inflammatory markers in patients with non-ST segment elevation acute coronary syndrome[J]. Chin Med J(Engl), 2006,119(1):32-36
7. Calin Popa, Mihai G. Netea, Piet L. C. M. van Riel, Jos W. M. van der Meer, and Anton F. H. Stalenhoef.The role of TNF-in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk.J. Lipid Res.,2007; 48:751-762
8. Daniel R. Meldrum. Tumor necrosis factor in the heart. Am J Physiol Regulatory Integrative Comp Physiol,1998; 274:577
9. May Azzawi and Philip Hasleton. Tumour necrosis factor alpha and the cardiovascular system:its role in cardiac allograft rejection and heart disease Cardiovasc Res,1999; 43:850-859
10.杨富国,刘革新,董果雄,等.黄芪甲苷对缺氧/复氧损伤人脐静脉内皮细胞与中性粒细胞黏附能力及细胞核转录因子κB表达的影响[J].中国组织工程研究与临床康复,2008,12(15)：2843-2846
11.高玉霞,黄国伟,刘英华,等.大豆异黄酮对体外血管内皮细胞氧化损伤保护作用的形态学观察[J].天津医科大学学报,2003,9(1)：39-41
12.王玉英,胡涛,贾国良,等.银杏叶提取物对人脐静脉内皮细胞增殖的影响[J].第四军医大学学报,2004,25(13)：1179-1181
13.陶波,李晓宁.蒲黄对动脉粥样硬化血管内皮损伤影响的实验研究[J].中西医结合心脑血管病杂志,2004,2(4)：222-223
14.周芳,李爱媛,谢金鲜,等.蒲黄抗鹌鹑高脂血症及动脉粥样硬化的实验研究[J].中国实验方剂学杂志,2006,12(8)：48-49
15.于长青.麦胚黄酮对实验老鼠血脂水平和脂质抗氧化作用影响的研究[J].中国食品学报,2001,2(1)：30
16.祝光礼,陈铁龙,陈启兰.黄芪失笑汤为主治疗气虚血瘀型冠心病.浙江中西医结合杂志[J].2006,16(10)：625-626
17.郑筱萸.中药新药临床研究指导原则(试行)[S].北京：中国医药科技出版社,2002：68-73
18.路志正.中医诊断学[M].北京：人民卫生出版社,1995：99-101
19.祝光礼,陈铁龙,陈启兰等.黄芪失笑汤为主治疗气虚血瘀型冠心病.浙江中西医结合杂志.2006,16(10)：625-626
20.杨利.邓铁涛教授“冠心三论”[J].湖南中医药导报,2004,10(6)：8-10
21.胡丹,姜亚军.从“痰瘀热毒”论治动脉粥样硬化[J].中医药信息,2011；28(1)：5-7
22.魏自民.活血化瘀法干预动脉粥样硬化的临床研究[J].光明中医,2011；26(2)：277-278
23.王永霞,胡宇才,朱明军.冠心病心绞痛中医证候分布相关性研究[J].世界中西医结合杂志,2008,3(12)：714-716
24.郭晓燕,王宏燕等.益气活血法治疗冠心病心绞痛50例[J].中国中医急症,2003,12(3)：264
25.邵莹.益气活血法治疗冠心病心绞痛临床观察[J].现代中西医结合杂志,2006,15(4)：422-423
26.刘剑刚,徐浩,董国菊,等.血府逐瘀口服液对冠心病心绞痛患者血小板活化分子表达的影响[J].长春中医药大学学报,2007,23(1)：29-31
27.卞云云,李萍.蒙古黄芪中黄酮类成分抗超氧阴离子活性研究及构效关系分析[J].中国药学杂志,2008,43(4)：256-259
28.尚正录,尚云等.黄芪保心汤预防实验性大鼠急性心肌缺血的研究[J].上海中医药杂志,2004,38(11)：47-50
29.严辉,陈佩东,丁安伟.蒲黄的炮制及其化学成分、药理作用的研究进展[J].江苏 中医药,2005,26(1)：59-61
30.陶波,李晓宁.蒲黄对动脉粥样硬化血管内皮损伤影响的实验研究[J].中西医结合心脑血管病杂志,2004,2(4)：222-223
31.杨琦,赵岩等.五灵脂的脂溶性成分GC-MS分析[J].中国现代中药,2010；12(10)：19-21
32.李强,陆蕴如,鲁学照,等.五灵脂化学成分的研究[J].中国药学杂志,1999,34(8)：514-516
33.祝光礼,范翠娟,陈铁龙,等.黄芪失笑散对鸡胚绒毛尿囊膜促血管生成实验研究[J].中华中医药学刊,2007,25(12)：2462-2464
34.祝光礼,周凡,陈铁龙.黄芪失笑散抗心肌缺血的实验研究[J].中华中医药学刊,2008,26(1)：2232-2234
35.祝光礼,范翠娟,陈铁龙,等.黄芪失笑散对实验大鼠梗死心肌内VEGF及bFGF mRNA基因表达的影响[J].中华中医药学刊,2008,26(1)：41-43
36.祝光礼,魏丽萍,方伟,等.黄芪失笑散对血管内皮功能的影响浙江临床医学[J].2008,10(8):1016-1018
37.曹建华.辛伐他汀对冠心病患者CRP、vWF、NO及ET的影响[J].中国老年学杂志,2010,30(4)：1138-1139
38. Buchman TG,Abello PA,Smith EH,et al.Induction of heat shock response leads to apoptosis in endothelial cells previously exposed to endotoxin[J].Am J Physiol,1993,265(1 Pt 2):H165-H170
39.Vita JA, Teasure CB, Nabel EG, et al.Coronary vasomotor response to acetylcholine relates to risk factor for coronary artery disease.Circulation,1990,81:491-497
40. Carswell EA,Old LJ,Kassel RL,et al.An endotoxin-induced serum factor that causes necrosis of tumors[J].Proc Natl Acad Sci USA,1975,72(9):3666-3670
41.Dinarello CA,Marqolis NH.Cytokine-processing enzymes. Stopping the cuts[J].Curr Biol,1995,5(6):587-590
42. Strieter RM,Kunkel SL,Bone RC.Role of tumor necrosis factor-a in disease states and inflammation[J].Crit Care Med,1993,21(10 Suppl):S447-S463
43.张敬芳,王光浩.TNF-a诱导HUVEC的损伤及其意义[J].荆门职业技术学院学报, 2002,17(6)：82-84
44.宋善俊,王林林,魏文宁.TNF-a刺激的人脐静脉内皮细胞组织因子表达及其分子机制[J].中国实验血液学杂志,2003,11(2)：124-127
45.吴军,李闪,朱建华,等.TNF-a对人脐静脉内皮细胞中NO和eNOS的影响[J].浙江医学,2006,28(8)：636-684
46. Min JK, Kim YM, Kim SW, et al. TNF-related activation-induced cytokine enhances leukocyte adhesiveness:Induction of ICAM-1 and VCAM-1 via TNF receptor-associated factor and protein kinase C-dependent NF-kappaB activation in endothelial cells. J fmmunol.2005,175(1):531-540
47. Pober JS, Cotran RS.Cytokines and endothelial cell biology Physiological Reviews,1990, 70427-70451
48. Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
49.刘学俭.失笑散小考[J].四川中医,1989(11)：13-14
50. Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
51.苏敏,钟翠平.动脉粥样硬化病变中黏附分子ICAM-1、VCAM-1及E-selectin的表达[J].第三军医大学学报,2009,31(11)：1066-1068
52.饶丹.黏附分子细胞间黏附分子-1/E-选择素与冠心病[J].心血管病学进展,2005,26(3)：278-281
53. Bitsch A, KleneW,Muttada L,et al.A longitudinal p rospectivestudy of soluble adhesion molecules in acute stroke[J].Stroke,1998,29:2129
54.高岩,夏蕾,白明.实验性大鼠肺血栓栓塞症中ICAM-1, P-选择素的变化及当归注射液的影响[J].中国微循环,2007,11(3)：176-179
55.陈良,张梅,李长江,等.复方丹参滴丸对动脉粥样硬化粘附因子的作用[J].中国动脉硬化杂志,2007,15(2)：101-104.
56.马丽萍,秦永文,郑兴,等.黏附分子与冠状动脉病变程度的相关性分析[J].基础医学与临床,2004,24(1)：44-47.
57.任强,李自成,巫少荣,等.血管细胞黏附分子-1在动脉粥样硬化中的作用[J].临床 荟萃,2004,19(7)：419-421.
58.隋广超,胡美浩.纤溶酶原激涪物抑制因子-1蛋白分子的结构与功能.国外医学生物分子学分册.1994.16(4)：172-176
59. Schneideerman J,Sawdey MS,Keeton Mr,et al.increased type 1 plsminogen activator inhibitor gene expreasion in atherosclerotic human arteries [J]. Proc Natl Acad Sci,1992,89: 6998～7002.
60. WenzIE. Dixit VM. Ubiqutin-mediated regulation of TNFR1 signaling[J]. Cytokine Growth Factor Rev,2008,19(3):313-324
61.Christman JW,lancaster LH,Blackwell TS.Nuclear factor kappa-B:a pivotal role in the system-inflammatory response and new target for the rapy.Intensive Care Med,1998,24(11):1131-1138
62. Sergei I. Grivennikov. Michael Karin. Dangerous liaisons:STAT3 and NF-KB collab oration and crosstalk in cancer [J]. Cytok Growth,2010,21(1):11-19
63. Varfolomeev E, Goneharov T, Fedorova AV, et al. C-IAP1 and c-IAP2 are critical mediators of tumor necrosis factor-a(TNF-a)-in-duced NF-KB activation[J]. Biol Chem, 2008,283(36):242-259
64.Wang P, Qiu W, Dudgeon C,et al. PUMA is direcfly activated by NF-KB and contributes to TNF-α-induced apoptosis[J]. Cell Death Differ,2009.16(9):192-202
65. Karin M,Greten FR. NF-KB:linking inflarmnation and immunity to cancer development and progression[J]. Nat Rev Immunol,2005,5(10):749-59
66. Hess J,Angel P,Schorpp-Kistner M.AP-1 subunits:quarrel and harmony among siblings[J]. J Cell Sci,2004,117(25):5965-5973
67. TouyzRM. Deschepper C, Park JB. Inhibition of mitogen activated protein/extracellular signal-regulated kinase improves endothelial function and attenuates Ang II-induced contractility of mesenteric resistance arteries from spontaneously hypertensive rats [J]. J Hypertens.2002,20(6):1127-1134
68. Kipsbidze N. Dangas G, Tsapenko M, et al. Role of the endothelium in modulating neointimal formation:vaseoloprotective approaches to attenuate restenosis after percutaneous coronary interventions[J]. J Am Coll Cardiol,2004,44(4):733-739
69. Dragomir E, Tircol M, Manduteanu I, et al. Aspirin and PPAR-alpha activators inhibit monocyte chemoattractant protein-1 expression induced by high glucose concentration in human endothelial cells [J]. Vaseul Pharmacol,2006,44(6):440-449
1.Carswell EA,Old LJ,Kassel RL,et al.An endotoxin-induced serum factor that causes necrosis of tumors [J].Proc Natl Acad Sci USA,1975,72(9):3666-3670
2.Dinarello CA,Marqolis NH.Cytokine-processing enzymes. Stopping the cuts[J].Curr Biol,1995,5(6):587-590
3.Aderem AA.How cytokines signal messages with in cells[J].J Infect Dis,1993,167(Suppl 1):S2-S7
4.Douzinas EE,Tsidemiadou PD,Pitaridis MT,et al.The regional production of cytokines and lactate in sepsis-related multiple organ failure [J].Am J Respir Crit Care Med,1997,155(1):53-59
5.Strieter RM,Kunkel SL,Bone RC.Role of tumor necrosis factor-a in disease states and inflammation[J].Crit Care Med,1993,21(10 Suppl):S447-S463
6.王今达,脓毒症：感染性M O D S的预防[J].中国危重病急救医学,1999,11(8)：453-455.
7.陶晓根.糖皮质激素抵抗与脓毒血症[J].中国危重病急救医学,2001,13(5)：307-309
8.Buchman TQAbello PA,Smith EH,et al.Induction of heat shock response leads to apoptosis in endothelial cells previously exposed to endotoxin[J].Am J Physiol,1993,265(1 Pt 2):H165-H170
9.赵克温.人脐带静脉血管内皮细胞的体外培养及其凋亡的研究[J].山东大学学报,2002,37(3)：268-271
10.金惠铭,刘清行,张国平,等.TNFa引起的微血管内皮细胞功能障碍及其分子机制[J].中国病理生理杂志,2000,16(10)：940-941
11.董红梅,喻伦银,贾宗智,等.TNFa对培养的脐静脉内皮细胞的损伤作用[J].临床与实验病理学杂志,2002,18(4)：401-403
12.Pober JS, Cotran RS.Cytokines and endothelial cell biology Physiological Reviews,1990, 70427-70451.
13.Meager A.Cytokine regulation of cellular adhesion molecule expression in inflammation.Cytokine Growth Factor Rev,1999,10(1):27-39
14.Sedgwick JD, Riminton DS.Cyster JG, et al.Tumor necrosis factora master-regulator of leukocyte movement.Immunol Today.2000,21 (3):110-113
15.Krown KA,Page MT,Nguyen C,et al.Tumor necrosis facor-a induced apoptsis in cardiac myocytes[J].JClin Invest,1996,98(12):2854-2865
16.周亚峰.肿瘤坏死因子对人血管内皮细胞凋亡的影响[J].苏州大学学报(医学版),2004,24(5)：620-623
17.Depraetere V,Golstein P.Fas and other cell death signaling pathways[J].Semin Immunol,1997,9(2):93-107
18.Duan H,Dixit VM.RAIDD is a new death adapt or molecule [J]. Nature,1997,385 (6611):86-89
19.Laster SM,Wood JG,Gooding LR.Tumor necrosis factor can induce both apoptosis and necrosic forms of cellysis[J] J Immunol,1988,141(8):2629-2634
20.姜华,张艳,王辰.益气活血复方含药血清对人脐静脉内皮细胞TLR4/NF-KB信号通路及TNF-a ICAM-1 mRNA表达的影响[J],辽宁中医药大学学报,2010(12)4：47
21.臧运华,唐明,郭瑞友等,溶栓颗粒含药血清对TNF-α损伤血管内皮细胞VEGF表达影响的研究[J],中国中医药科技,2011,18(1)：15-16
22.祝光礼,方小江,彭哲,等.冠心合剂对人脐静脉内皮细胞分泌血管生长因子bFGF的影响[J].浙江中西医结合杂志,2011,21(9)：596-598
23.祝光礼,魏丽萍,方伟,等.黄芪失笑散对实验大鼠主动脉和OX-LDL诱导下人脐静脉内皮细胞ICAM-1、VCAM-1 mRNA基因表达的影响[J].中华中医药学刊,2008,26(10)：2095-2098
24.尚改萍,文志斌,李俊成等,补阳还五汤对TNFa诱导血管内皮细胞释放vWF及表达组织因子的影响[J],湖南医科大学学报,2000,25(2)：129-131
25.黄建林,林灼锋,罗敏琪等,青藤碱抑制TNF-α诱导人脐静脉内皮细胞VCAM-1表达[J],中国病理生理杂志,2007,23(4)：634-638
26.叶泽兵,汪凡军,张刚庆等,黄芪注射液对TNF-α诱导人脐静脉内皮细胞炎症因子的影响及其分子机制[J],南方医科大学学报,2009,29(6)：1137-1140
27.游洋,段岩,张效林等,黄芪水提取物对TNF-α诱导的小鼠动脉内皮细胞VCAM-1表达的影响[J],中国动脉硬化杂志2011(19)3：187-190
28.刘蓓,朱海燕,高永红,黄芪多糖对TNF-α诱导心脏微血管内皮细胞黏附分子基因转录及p38MAPK信号通路的影响[J],世界中医药,2011(6)3：263-265