桂芍通络对兔动脉粥样硬化早期外膜滋养血管新生及氧化应激水平的干预作用

Influence of Guishaotongluo on angiogenesis of adventitial vasa vasorum and oxidative stress in early stage of atherosclerosis

目的观察桂芍通络对兔动脉粥样硬化早期外膜滋养血管新生及氧化应激的影响。方法 84只家兔随机分为正常组、高脂组、外膜损伤组、桂芍通络高剂量(GH)组、桂芍通络中剂量(GM)组、阿托伐他汀(ATO)组、通心络(TXL)组,每组各12只。正常组给予普通饲料,高脂组给予高脂饲料喂养建立早期高脂血症模型,外膜损伤组及各用药组实施高脂饮食复合右侧颈动脉硅胶管包裹术。GH组给予4.16 g(生药)·kg^(-1)·d^(-1)灌胃,GM组给予2.08 g(生药)·kg^(-1)·d^(-1)灌胃,ATO组给予阿托伐他汀2.5 mg·kg^(-1)·d^(-1)灌胃,TXL组给予通心络超微粉0.6 g·kg^(-1)·d^(-1)灌胃,连续给药4周后取材,生化法检测各组血脂4项水平变化;HE染色观察内中外膜病理形态学变化;检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)表达水平;荧光原位杂交检测NADPH氧化酶亚单位p22phox和gp91phoxmRNA的定位与表达;Western blot检测颈动脉组织VEGF、VEGFR蛋白表达情况。结果与正常组相比,高脂组血清总胆固醇(TC)、甘油三酯(TG)及低密度脂蛋白(LDL-C)水平明显升高(P<0.01);外膜损伤组外膜新生滋养血管及VEGF、VEGFR-2蛋白表达均明显增加(P<0.01),与外膜损伤组比较,GH组、GM组、ATO组及TXL组术侧颈动脉损伤及滋养血管新生程度、颈动脉外膜NADPH氧化酶亚单位p22phox和gp91phoxmRNA均有不同程度减轻;SOD、T-AOC活力升高,MDA含量、颈动脉外膜VEGF、VEGFR蛋白表达水平降低(P<0.05或P<0.01)。结论桂芍通络具有抑制兔动脉粥样硬化早期血管外膜滋养血管新生的作用,其机制可能与提高血管本身及外膜的抗氧化能力有关。

Aim To observe the effect of Guishaotongluo(GSTL) on the angiogenesis of vasa vasorum and oxidative stress in the early stage of atherosclerosis.Methods The rabbits(n=84) were randomly divided into 7 groups(n=12):control group,high-fat group,adventitial injury group,GSTL high(GH) and medium(GM) dose group,atorvastain group(ATO),and Tongxinluo group(TXL).The normal group was fed with common foodstuffs,and high-fat foodstuffs for the high-fat group to establish an early model of hyperlipidemia,and all the other groups were fed with highfat diet combined with carotid artery cannula to build early atherosclerosis carotid artery injury rabbit models.The GSTL high and medium dose was given Guishaotongluo ultrafine powder 4.16,2.08 g·kg^(-1)·d^(-1) respectively.The atorvastain group and Tongxinluo group were given suspension of atorvastain solution 2.5mg·kg^(-1)·d^(-1),Tongxinluo supermicro powder 0.6 g·kg^(-1)·d^(-1).All groups were treated with gastric perfusion for 4 weeks.Biochemical method was applied to detect blood lipid change.HE staining was used to observe the pathological morphology of intima-media.Aactivity of serum superoxide dismutase(SOD),malondialdehyde(MDA) content and the total antioxidant capacity(T-AOC) in artery serum were detected.NADPH subunits p22 phox mRNA,gp91 phoxmRNA in carotid arteries were located and semi-quantitated by fluorescence in situ hybridization.The expression of VEGF,VEGFR-2 in the carotid artery adventitia was detected by Western blot.Results Compared with normal group,the contents of TC,TG and LDL-C were significantly increased,and VEGF,VEGFR-2 protein levels were remarkly increased in high-fat and adventitial injury group.The carotid artery injuries,the degree of angiogenesis of vasa vasorum and NADPH subunits p22 phox,gp91 phoxmRNA in adventitia tissue of the GH,GM,ATO and TXL group were milder in varying degrees compared with those of the vasa injury group.Also the activity of SOD,T-AOC increased,while MDA content,VEGF,VEGFR-2 protein levels were remarkly decreased(P<0.5 or P<0.01).Conclusions GSTL can inbibit adventitial neovascularization in the early stage of atherosclerosis,and its mechanism might be related to the increase of total antioxidant capacity of the vascular system and adventitia tissue.