基于血流剪切力与炎性反应探讨当归补血汤治疗动脉粥样硬化的作用机制

Exploration on the Mechanism of Danggui Buxue Decoction in the Treatment of Atherosclerosis Based on Fluid Shear Stress and Inflammatory response

目的基于血流剪切力(FSS)与炎性反应探讨当归补血汤对动脉粥样硬化(As)家兔的作用机制。方法将60只新西兰家兔随机分为假手术组、模型组、阿托伐他汀组(1.7 mg·kg^(-1))及当归补血汤高、中、低剂量组(6、3、1.5 g·kg^(-1)),每组10只。采用右侧颈总动脉套管术结合高脂饲料诱导建立FSS异常的As家兔模型,连续造模4周。造模成功后,给药组分别按照设定剂量灌胃给药,假手术组灌胃等量生理盐水,每日1次,连续灌胃12周。采用右侧颈总动脉超声检测血流动力学指标及计算FSS变化;检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)水平;测定全血黏度、血浆黏度(ƞp)及红细胞变形指数(TK);培养鉴定外周血内皮祖细胞(EPCs)及检测其集落数、迁移力、成小管力;ELISA法检测血管内皮组织中细胞间黏附分子1(ICAM-1)水平;HE染色法观察血管内皮组织病理学变化;免疫荧光法检测血管组织中核转录因子κB(NF-κB)、血管细胞黏附分子1(VCAM-1)蛋白表达水平。结果与模型组比较,当归补血汤各剂量组家兔的收缩期峰值流速(Vs)、舒张期血流速度(Vd)、平均血流速度(Vm)及FSS明显提高(P<0.05),颈动脉管腔未见明显狭窄,内膜平滑,未见粥样斑块形成;当归补血汤各剂量组的血清TC、TG、LDL-C水平明显降低(P<0.05),中剂量组的HDL-C水平明显升高(P<0.05);当归补血汤中、高剂量组的全血黏度、血浆黏度明显降低(P<0.05),中剂量组的红细胞变形指数明显升高(P<0.05);当归补血汤各剂量组的EPCs集落明显增加(P<0.05),EPCs迁移力及成小管力明显增强(P<0.05);当归补血汤各剂量组家兔血管内皮组织的NF-κB、VCAM-1蛋白表达水平及ICAM-1含量明显降低(P<0.05)。结论当归补血汤可能通过改善血流动力学指标,提高FSS,改善脂质代谢水平、全血黏度及外周血EPCs功能,抑制血管内皮炎症损伤,从而发挥抗As的作用。

Objective To investigate the mechanism of action of Danggui Buxue Decoction in rabbits with atherosclerosis(As)based on fluid shear stress(FSS)and inflammatory response.Methods Sixty New Zealand rabbits were randomly divided into the sham-operated group,the model group,the Atorvastatin group(1.7 mg·kg^(-1))and the high-,medium-and low-dose of Danggui Buxue Decoction groups(6,3 and 1.5 g·kg^(-1)),10 rabbits in each group.The As rabbit model of FSS abnormality was established by right common carotid artery cannulation combined with high fat feed induction for 4 continuous weeks.After successful modelling,the drug administration group was administered by gavage at the set dose,and the sham-operated group was gavaged with the same amount of saline once a day for 12 continuous weeks.The right common carotid artery ultrasound was used to detect haemodynamic parameters and calculate FSS changes;serum TC,TG,LDL-C and HDL-C levels were detected;whole blood viscosity,plasma viscosity(ƞp)and erythrocyte deformation index(TK)were measured;peripheral blood endothelial progenitor cells(EPCs)were identified by culture and their colony number,migration force and tubule-forming force were detected.The levels of ICAM-1 in vascular endothelial tissues were detected by ELISA method,and the protein expression levels of NF-κB and VCAM-1 in vascular tissues were measured by immunofluorescence method.Results Compared with the model group,the peak systolic flow velocity(Vs),diastolic flow velocity(Vd)and FSS of rabbits in each dose group of Danggui Buxue Decoction were significantly increased(P<0.05),the carotid lumen was not significantly narrowed,the intima was smooth,and no atheromatous plaque formation was observed;the levels of serum TC,TG and LDL-C were significantly decreased in each dose group of Danggui Buxue Decoction(P<0.05),and HDL-C levels were significantly increased in the meidum-dose group(P<0.05).The whole blood viscosity and plasma viscosity were significantly decreased in the Danggui Buxue Decoction group(P<0.05),and the erythrocyte deformation index was significantly increased in the meidum-dose group(P<0.05);the colonization of EPCs was significantly increased in the Danggui Buxue Decoction group(P<0.05),and the migration force and tubule-forming force of EPCs were significantly enhanced in the Danggui Buxue Decoction group(P<0.05).The protein expression levels of NF-κB,VCAM-1 and ICAM-1 in vascular endothelial tissues of rabbits in each dose of Danggui Buxue Decoction group were significantly decreased(P<0.05).Conclusion Danggui Buxue Decoction can exert anti-As effects by improving hemodynamic indexes,increasing FSS,improving lipid metabolism levels,whole blood viscosity and the function of peripheral blood EPCs,and inhibiting inflammatory damage to the vascular endothelium.