动脉粥样硬化模型主动脉血管细胞黏附分子1mRNA表达量与益气活血法干预的量效关系(英文)

Dose-effect relationship between the expression of aortic vascular cell adhesion molecule-1 mRNA and qi-benefiting and blood-activating intervention on atherosclerosis

背景:祖国医学中,本虚标实、气虚血瘀为动脉粥样硬化辩证论治常见证型之一,益气活血剂重用益气药物,然而益气、活血药的最佳权重关系需进一步研究。目的:观察益气活血法对动脉粥样硬化模型主动脉血管细胞黏附分子1mRNA表达量的影响,分析益气活血药黄芪、三七的交互作用和量效关系。设计:随机对照动物实验。单位:山西医科大学。材料:实验于2005-04在山西医科大学完成。健康雄性Wistar大鼠60只。槲皮素(由陕西彗科植物有限公司提供,批号20041112);三七总皂甙(由昆明雅阁臣药业有限公司提供,批号20050118);川芎嗪(由预新国际龙源药业有限公司提供,批号20041204)。方法:造模:Wistar大鼠饲喂动脉粥样硬化饲料(蛋黄粉10%、猪油5%、胆盐0.5%、基础饲料85%),喂服3个月。分组及给药:60只雄性Wistar大鼠,适应性喂养3d后,随机抽取8只做为正常对照组,继续饲喂普通饲料,其余饲喂动脉粥样硬化饲料。3个月后随机抽取4只进行血脂测定及主动脉组织标本检测,验证造模成功,再将其余48只随机分为6组:①黄芪治疗组:造模成功后给予槲皮素0.1g/(kg·d)灌胃。②三七治疗组:造模成功后给予三七总皂甙0.1g/(kg·d)灌胃。③黄芪三七2∶1治疗组:造模成功后给予槲皮素0.1g/(kg·d),三七总皂甙0.05g/(kg·d)灌胃。④黄芪三七3∶1治疗组:造模成功后给予槲皮素0.15g/(kg·d),三七总皂甙0.05g/(kg·d)灌胃。⑤川芎嗪治疗组:造模成功后给予川芎嗪0.2g/(kg·d)灌胃。⑥正常对照组:饲喂普通饲料。⑦模型组:造模成功后改喂普通饲料。用药30d后采用RT-PCR法检测主动脉中血管细胞黏附分子1mRNA的相对表达水平。取尾静脉血2mL测血清总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平。主要观察指标:①主动脉中血管细胞黏附分子1mRNA的相对表达水平。②血脂水平。结果:60只雄性Wistar大鼠参加实验,51只雄性Wistar大鼠进入结果分析,其中治疗组大鼠因为灌胃不慎死亡3只,模型组大鼠意外死亡2只。①高脂饮食可诱导大鼠形成动脉粥样硬化模型,模型组的血脂水平明显高于正常组(P<0.05),各干预组血清总胆固醇、三酰甘油、低密度脂蛋白胆固醇水平低于模型组(P<0.05)。②正常对照组无血管细胞黏附分子1mRNA表达。各干预组血管细胞黏附分子1mRNA表达量明显低于模型组。黄芪三七3∶1治疗组的血管细胞黏附分子1mRNA表达量水平最低(0.42±0.02),与其他干预组比较,差异有显著性意义(P<0.05)。结论:益气活血药黄芪、三七可以下调主动脉组织血管细胞黏附分子1mRNA的表达量,降低血脂,具有抗动脉粥样硬化作用,同时,联合应用比单独应用效果更好,并且在一定范围内,随着益气药物量的增加,抗动脉粥样硬化作用增强。

BACKGROUND： In traditional Chinese medicine, both deficiency as primary and excess as secondary and deficiency of qi and blood stasis are common symptoms in dialectical Iogy of atherosclerosis （AS）. Therefore, qi-benefiting drugs are the main component of qi-benefiting and blood-activating intervention. However, the best dose relationship between qi-benefiting and blood-activating drugs needs to be further studied. OBJECTIVE ： To observe the effect of qi-benefiting and blood-activating intervention on the expression of aortic vascular cell adhesion molecule-1mRNA （VCAM-1mRNA） in AS models and analyze dose-effect relationship between astragalus and sanchi. DESIGN： Randomized control animal study. SETTING： Shanxi Medical University. MATERIALS： The experiment was carried out in the Shanxi Medical University in April 2005. A total of 60 healthy male Wistar rats were selected in this study. The main reagents were quercetin （Shaanxi Huike Plant Co., Ltd., batch number： 20041112）, saponins of panax notoginseng （PNS, Kunming Yagechen Pharmaceutical Co., Ltd., batch number： 20050118） and ligustrazine （Yuxin Guoji Longyuan Pharmaceutical Co., Ltd., batch number： 20041204）. METHODS ： Model establishment： Wistar rats were administrated AS feeds （including 10% yolk powder, 5% lard, 0.5% bile salt and 85% basic feed） for 3 months. Grouping and administration： At three days after suitability feeding, 8 rats were randomly selected, regarded as the normal control group and given general feeds, and other 52 rats were fed with AS feeds. Three months later, 4 rats were randomly selected for the measurements of lipid and aortic tissue. And then, the models were established successfully. In addition, 48 rats were randomly divided into 6 groups. ① Astragalus treatment group： Rats were per/used with 0.1 g/（kg·d） quercetin. ② Sanchi treatment group： Rats were per/used with 0.1 g/（k·d） PNS. ③ 2：1 of astragalus/sanchi treatment group： Rats were per/used with 0.1 g/（kg·d） quercetin and 0.05 g/（kg·d） PNS. ④ 3：1 of astragalus/sanchi treatment group： Rats were per/used with 0.15 g/（kg·d） quercetin and 0.05 g/（kg·d） PNS. ⑤ Ligustrazine treatment group： Rats were per/used with 0.2 g/（kg·d） ligustrazine （Yuxin Guoji Longyuan Pharmaceutical Co., Ltd.）. ⑥ Normal control group： Rats were fed with general feeds. ⑦ Model group： Rats were fed with general feeds after successful model estalishement. Thirty days after administration, relative expression of VCAM-1 mRNA in aorta was measured with reverse transcription polymerase chain reaction （RT-PCR） technique; moreover, 2 mL venous blood was collected from tail to measure the levels of serum total cholesterol （TC）, triacylglycerol （TG）, high density lipoprotein cholesterol （HDL-C） and low density lipoprotein cholesterol （LDL-C）. MAIN OUTCOME MEASURES： ① Relative expression of VCAM-lmRNA in aorta; ② level of lipid. RESULTS ： Among 60 male Wistar rats, three rats in the treatment group died because of perfusion and two rats in the model group died due to accident; therefore, a total of 51 rats were involved in the final analysis. ① High-lipid diet could promote the formation of AS models. Level of lipid was higher in the model group than that in the normal control group （P 〈 0.05）, and leves of serum TC, TG and LDL-C were lower in the interventions group than those in the model group （P 〈 0.05）. ② Expression of VCAM-lmRNA was not found in the normal control group. Expression of VCAM-lmRNA was lower in the intervention groups than that in the model group. Expression of VCAM-lmRNA （0.42±0.02） was the lowest in 3：1 of astragalus/sanchi treatment group, and there were significant differences as compared with other intervention groups （P 〈 0.05）. CONCLUSION： Astragalus and sanchi, a main component of qi-beneflting and blood-activating herbs, can down-regulate the level of lipid and resist AS; meanwhile, the combination of them is superior to the single application; in addition, with the increasing deal of qi-benefiting drugs, the function against AS is strengthened.