通心络联合阿托伐他汀、阿司匹林对家兔动脉粥样硬化早期血管外膜滋养血管新生的干预作用

摘要

目的：观察通心络联合阿托伐他汀、阿司匹林对家兔动脉粥样硬化早期颈动脉外膜滋养血管新生的影响。方法72只健康♀♂各半新西兰白兔随机分为对照组、模型组、通心络(TXL)组、阿托伐他汀(ATO)组、阿司匹林(ASP)组、金三角(ATS)[1]组,各12只。对照组给予普通饲料、模型组及各用药组家兔均实施单侧颈动脉硅胶管包裹术复合高脂饲料喂养,TXL 组给予通心络超微粉混悬液0．3 g·kg-1· d-1灌胃,ATO 组给予阿托伐他汀2．5 mg·kg-1·d-1灌胃, ASP 组给予阿司匹林12 mg·kg-1·d-1灌胃,ATS 组给予通心络超微粉混悬液0．3 g·kg-1·d-1加阿托伐他汀2．5 mg ·kg-1·d-1加阿司匹林12 mg·kg-1·d-1灌胃,连续给药4周后取材,HE 染色观察颈动脉内中膜的变化；免疫组化法检测颈动脉外膜CD34表达情况；微球检测颈动脉微血管血流量的变化；RT-PCR 和Western blot 检测颈动脉组织中 VEGF、VEGFR-2基因和蛋白表达情况。结果与对照组比较,模型组VEGF、VEGFR-2基因和蛋白表达以及微血管血流量明显增强(P <0．01)。与模型组比较,各用药组VEGF、 VEGFR-2基因和蛋白表达以及微血管血流量明显减弱(P <0．01,P <0．05)。与其他用药组比较,ATS 组VEGF、VEGFR- 2基因和蛋白表达以及微血管血流量明显减弱(P <0．01,P <0．05)。与ASP 组比较,TXL 组、ATO 组VEGFR-2蛋白表达均明显降低(P <0．05),ATO 组微血管血流量减少明显(P <0．05)。TXL 组与ATO 组两组间比较差异无显著性(P >0．05)。VEGF、VEGFR-2基因和VEGF 蛋白表达组间比较无统计学意义(P >0．05)。各用药组颈动脉外膜CD34表达减少。结论“ATS”方案有助于减少动脉粥样硬化早期颈动脉VEGF、VEGFR-2表达,抑制血管外膜滋养血管新生,减少促炎物质进入血管中膜、内膜,延缓动脉粥样硬化进程。%Aim To observe the effect of a treatment proposal named “Golden Triangle” ( Atorvastatin, Tongxinluo,and Aspirin) on the vasa vasorum angio-genesis of early atherosclerosis lesions in rabbits carotid artery. Method Seventy-two healthy New Zealand rabbits with half males and half females were divided into 6 groups randomly ( n =12 ):control group, model group, Tongxinluo group ( TXL ) , atorvastatin group ( ATO ) , aspirin group ( ASP ) , golden triangle group ( ATS) . The control group was fed with common feed-stuff, and all the other groups′ right carotid arteries were equipped with the silicone tube,and were then fed with fatty feedstuff. The Tongxinluo group, the Atorvas-tatin group and the Aspirin group were given suspen-sion of Tongxinluo supermicro powder(0. 3 g·kg-1 · d-1 ) , Atorvastatin ( 2. 5 mg · kg-1 · d-1 ) and Aspirin (12 mg·kg-1·d-1),the golden triangle group were given suspension of Tongxinluo supermicropowder (0. 3g·kg-1 ·d-1),atorvastatin(2. 5 mg·kg-1 · d-1 ) and Aspirin ( 12 mg · kg-1 · d-1 ) . All the groups were fed with medicine for 4 weeks. Tissue slice of carotid artery was stained with HE and observed un-der light microscope. The change of blood liquid was detected by biochemical assay. Immunohistochemical staining was used to detect the protein expression of CD34 around the carotid artery adventitia. Color micro-sphere method was used to detect the blood flow vol-ume change of the cartoid artery microvascular. VEFG, VEGFR-2 gene and protein expression in the cartoid artery were detected by Real-time PCR and Western Blot. Result Compared with the control group,the content of VEGF, VEGFR-2 gene and pro-tein expression and the microvascular blood flow vol-ume of cartoid artery microvascular in the model were significantly increased ( P <0. 01 ) . But those in each drug group were lighter than those in the model group (P<0. 01,P <0. 05). In the ATS group,the content of VEGF, VEGFR-2 gene and protein expression and the microvascular blood flow volume of cartoid artery microvascular were lower than those in the TXL, ATO and ASP group ( P <0. 01 , P <0. 05 ) . Compared with the ASP group,the content of VEGFR-2 protein expres-sion was significantly decreased(P<0. 01)in the TXL and ATO group. VEGF,VEGFR-2 gene and protein ex-pression in different subgroups showed no significant difference( P >0. 05 ) . The content of CD34 was de-creased in TongxinLuo group,atorvastatin group,aspirin group and ATS group. Conclusion The ATS project can reduce the expression of VEGF,VEGFR-2, inhibit the vasa vasorum angiogenesis and decrease proinflam-matory substances in the tunica media and intima of vascular wall. It plays an important role in intervening in the process of AS.