【作者】 葛玉霞；

【导师】 段云友；

【作者基本信息】 第四军医大学 ， 影像医学与核医学， 2004， 硕士

【摘要】 血管内皮功能障碍是动脉粥样硬化的最敏感指标之一，它出现于血管形态学改变之前。高同型半胱氨酸血症(HHCY)是心血管疾病的一个新的独立的危险因素，但是其损伤血管内皮功能的机制尚不明确。本研究通过建立高同型半胱氨酸血症动物模型，探讨其导致内皮功能障碍的机理。复方丹参滴丸可明显降低冠状动脉事件的再发生危险和死亡率。但有关中药复方丹参滴丸对血管内皮功能的作用尚少见报道。本研究旨从基础和临床的角度研究血管内皮功能损伤的机理以及早期干预内皮功能损伤在防治冠心病中的价值。目的：(1)检测HHCY导致内皮功能损伤时，兔血中NO、ET以及GSH-PX活性和sVCAM-1的变化，从而观察氧化应激在活体内皮功能损伤中的作用：(2)评价抗氧化剂Vitc对HCY导致的兔血管内皮功能损伤是否具有保护作用。(3)探讨复方丹滴丸对具有冠心病危险因素人群血管内皮依赖性舒张功能的保护作用。 材料和方法：(1)实验一：将兔随机分为两组，每组9只，分别灌服3.3g／kg体重的L-蛋氨酸(M组)，等量自来水(N组)。然后于0、4、8、12、24小时检测其血浆中NO、ET以及GSH-PX的活性和sVCAM-1的含量，同时利用高分辨力超声检测其EDD、EID，24小时后电镜观测兔冠状动脉超微结构的第四军医大学硕士学位论文 改变。(2)实验二:将兔随机分为3组，每组9只，分别喂食正常饲料(N组)， L一蛋氨酸(0 .8叭g体重“天)饲料(M组)，L一蛋氨酸十巧tc(1.0叭g体重·天)饲 料MC组，然后于第20天、40天、60天、80天采血检测NO、ET、GsH一X 活性和sVCAM一1变化，同时利用高分辨力超声检测其EDD、ED〕。(3)实 验三:入选60例具有冠心病危险因素的患者，随机分为2组，治疗组(3O 例)每日口服复方丹参滴丸20粒，每日两次，对照组(3O例)均未服用任 何药物，治疗时间共6个月，同时利用高分辨力超声分别于O天、30天、90 天、180天检测胧动脉EDD和EID。 结果 (l)急性扣旺CY时，M组兔灌服蛋氨酸。、4、8、12和24小时后，骼外动 脉EDD分别为:1 3.9肚1.96%、3.5肚1.61%、一1.53士3.01%、一2.4矢1.83%和 一5，03士l‘62%，较对照组显著降低;而EID无明显改变。 (2)急性HHCY时，生化检测指标的变化:①血浆中NO的变化:M组兔 血浆中NO在上述各时间点与对照组相比(208.31士22.45切吐O比)均明显降 低，分别为183.5余38.10 umo讥、127.13土21.23 umo讥、115.2矢10.04切mo讥、 102.24士15.01 uzno比和97.35土31.12 umo况;②血桨ET的变化:M组兔 血浆ET在上述各时间点与对照组(174.1肚24.巧p留血l)相比明显升高 (P<o.ol)，分别为190.37土46.28 pg/inl、258.4肚71.36州刘、368.43士51.27 四ml、447.3矢37.21p留ml和399.2妊41.37p创ml:③血浆中GsH.pX的活 性及sVCAM一1的浓度变化:血浆中GSH·PX的活性及sVCAM一1的浓度与 对照组无明显差异。 (3)兔冠状动脉超微结构的改变:急性HHCY，透射电镜下，兔冠状动脉内 皮细胞均出现脱落，线粒体肿胀，内质网扩张、内弹力板断裂、平滑肌细胞 通过断裂的内弹力板向内皮下侵入。 (4)慢性H日CY时，随时间点推移，MC组和M组EDD均明显下降，EID 3第四军医大学硕士学位论文在60天后开始下降。其中80天时，MC组和M组的EDD分别为2.7狂3.46%、一6.46划.33%，均较对照组(13.1肚2.45%)明显降低，而且此两组之间也差异显著(P<0 .01)。EID(MC组:1 8.75士9.41%和M组:14.3肚4.10%)亦较对照组(25.03士2.87%)明显降低(P<0 .05)，但此两组之间无明显差异。(5)慢性HHCY时，生化检测指标的变化:①血浆中NO的变化:M组和MC组血浆中NO水平随时间推移，均明显下降。其中80天时，M组和MC组NO分别为25 1 .37士16.49 umol几和309.34土61.28 umo垅，较对照组(359.51士25.37um。巩)均明显降低(P<0 .05):MC组和对照组之间无明显差异。②血浆ET的变化:M组和MC组血浆中ET水平随时间推移，均明显上升。其中80天时，M组和MC组血浆ET分别为271料1 .38pg/inl和238.5妊23.12pg加l，较对照组(180.21士21.35p留ml)明显升高(p<0.05);M组和Me组之间无明显差异。③血浆中GSH一PX的活性及sVCAM一1的浓度变化:M组和MC组血浆中GSH一PX活性随时间推移明显下降，sVCAM一1浓度上升。其中80天时，M组和MC组GSH一PX活性分别为1 21 .4肚8.98活力单位和157，48月1.22活力单位，与对照组(l 68.4社37.25活力单位)相比明显降低 (P<0 .05);而MC组和对照组之间无明显差异。④M组和MC组血清中sveAM一l浓度分别为0.0951士0.0095 ng/而和0.0542土0.ooZn创d，较对照组(0.0045士o，0058 ng/IYil)均明显升高(P<0.01.)，MC组亦较M组明显降低 (P<0 .05)。(6)复方丹参滴丸对人体内皮功能损伤的保护作用 复方丹参滴丸可显著改善胧动脉的血管内皮功能，其中服药180天时EDD为1 3 .40士0.53%，较对照组(4.20士1 .26%)明显增加(P<0 .01)，且由血流介导的胧动脉血流量亦较对照组明显增加(487.40士63.42更多还原

【Abstract】 Vascular endothelial dysfunction is one of the most sensitive indicators for atherosclerosis, which displays prior to the changes of vascular morphology. Hyperhomocysteinemia is a new independent risk factor for cardiovascular disease, but the mechanism of the vascular injury seen in hyperhomocysteinemia is unknown. This study investigated its mechanism in endothelial dysfunction by establishing animal model of hyperhomochysteinemia. Compound danshen dripping pills could obviously decrease the reoccurance and mortality rate of cardiac attack. However there was no report on the effect of the compound danshen dripping pills on vascular endothelial function. This study is to investigate the mechanism of vascular endothelial dysfunction from basic and clinical point of view and to evaluate the significance of early intervention on vascular endothelial dysfunction in prevention and cure of coronary artery disease.Objectives: 1. Through observation of the changes of NO ET and activity ofGSH-PX> VCAM-1 in plasma of HHCY rabbits to study the effects of oxidant stress in endothelial dysfunction; 2. To evaluate the effect of antioxidant ascorbic acid(Vitc) in protecting hyperhomochysteinemia-induced endothelial dysfunction in rabbits; 3. To observe effect of the compound Dan shen dripping pills on vascular endothelium functionMaterials and methods: 1. Experiment one: Eighteen rabbits were randomized into two groups (n=9 per group), methionine group (M group) were poured L-methionine (3.3g/kg weight)into stamach. NO. ET and activity of GSH-PX. sVCAM-1 in plasma were examined at 0. 4. 8. 12. 24hour,at same time, EDD. EID were detected by high resolution ultrasound and external ilial arteries were observed by trandelectron microscope after 24 hours; 2. Experiment two: Rabbits were randomized into three groups (N=9 per group), normal control group (N group)received normal diet, methionine group(M group)received methionine-rich diet consisted of L-methioine 0.8g/kg weight day, methionine+Vitc group (MC group) received diet consisted of L-methioine 0.8g/kg weight day and Vitc 1.0g/kg weight day. Then NO, ET and activity of GSH-PX. sVCAM-1 in plasma were examined at 20. 40. 60. 80day. At same time EID. EDD were examined by transmission eleciron microscope(HRUS) after 80days; 3. Experiment three: 60 patients associating with CAD risk factors in our study. They were randomly divided into two groups, Control group(n=30)who don’t use any drug, treatment group(n=30) took composite Dan Shen pill 20-pill, twice every day, for sixmonths. With high-resolution ultrasound examine EDD. EID of the brachial artery at Oday. 30day. 90day. 180day.Results 1. The changes of EDD in acute hyperhomocysteinemia. EDD of external iliac artery in M group was time-dependent reduced significantly(Ohour, 13.98+1.96%; 4hour, 3.50+1.61%; 8hour, -1.53+3.01%; 12hour, -2.49+1.83%; 24hour, -5.03+ 1.62%), compared with that in control group; but EID was not affected. 2. Biochemical changes in acute hyperhomocysteinemia (1) NO in plasma: NO in plasma of M group was reduced significantly than that of control group (N group: 208.3 l+22.45umol/L vs M group Ohour. 183.56+38.10 umol/L: 4hour: 127.13+21.23 umol/L: 8hour: 115.29+10.04 umol/L; 12hour: 102.24+15.01 umol/L; 24hour: 97.35+31.12umol/L);(2)ETin plasma: ET in plasma of M group rabbits was increased significantly (N group: 174.12+24.15pg/ml vs M group : Ohour: 190.37+46.28 pg/ml ; 4hour: 258.42+71.36 pg/ml; 8hour: 368.43+51.27 pg/ml; 12hour: 447.39+37.21 pg/ml; 24hour: 399.29+41.37 pg/ml: P<0.01). (3) Changes of GSH-PX and sVCAM-1: There was no difference in GSH-PX sVCAM-1 between M and N group. 3. Endothelian function changes in chronic hyperhomocysteinemia: EDD decreased significantly in both MC and M groups with time and EID began to decrease 60 days later. After 80 days, EDD of external iliac arteries in MC group and M group was 2.77+3.46% and -6.46+1.33% and reduced significantly compered with mat in N group (13.10+2.45%, p<0.01). EDD in M group reduced significantly than that in MC group, while更多还原