当归补血汤对兔动脉粥样硬化模型骨髓内皮祖细胞的影响

Influence of Dang Gui Bu Xue Tang on spinal endothelial progenitor cells in rabbit model of atherosclerosis

目的观察当归补血汤对兔动脉粥样硬化(atherosclerosis,As)模型骨髓内皮祖细胞(endothe-lial progenitor cells,EPCs)的影响,探讨当归补血汤防治As的可能作用机制。方法新西兰大白兔25只,免疫损伤联合高脂饮食法4周后形成As模型,模型动物按体重随机分为5组,每组5只。空白组灌胃等量蒸馏水,辛伐他汀组灌胃辛伐他汀水悬液1.7 mg/kg,当归补血汤高、中、低剂量组分别灌胃当归补血汤63、、1.5 g/kg,每日1次2,周后取各组兔骨髓,密度梯度离心法分离单个核细胞,培养7 d后将贴壁细胞与Dil标记的乙酰化低密度脂蛋白和FITC标记的荆豆凝集素1共同孵育鉴定EPCs。计算细胞集落形成数,MTT法检测增殖力,黏附实验检测黏附力t,ranswell小室检测迁移力,体外血管生成试剂盒检测成小管力。结果与空白组比较,当归补血汤高、中剂量组及辛伐他汀组骨髓EPCs集落数及增殖力、黏附力、迁移力、成小管力均提高(P<0.05),低剂量组则无明显变化。结论当归补血汤抗As的作用与促进EPCs数量和活性具有密切的关系。

Objective To observe the influence of Dang Gui Bu Xue Tang on spinal endothelial progenitor cells （EPCs） in rabbit model of atheroselerosis, and discuss the possible mechanism of Dang Gui Bu Xue Tang in treating atherosclerosis. Methods The model of atherosclerosis was established by using immune injury and fatty diet for 4 weeks in New Zealand rabbits （n = 25 ）. All modeled rabbits were divided into 5 groups （each n = 5） according to the body weight. The blank group was intragastrically given distilled water, simvastatin group, aqueous suspension of simvastatin （1.7 mg/kg）, and high-dose group （6 g/kg）, mid-dose group （3 g/kg） and low-dose group （1.5 g/kg）, Dang Gui Bu Xue Tang once a day for two weeks. The mononuelear cells were isolated from rabbit bone marrow by using density gradient centrifugation, which were cultured for 7 days and then adherent cells were cultured with Dil-ac-LDL and FITC-UEA-1 together for identifying EPCs and calculating the number of EPCs colony forming. The proliferation and adhesion were detected by using MTT method, adhension, by using adhension determination, migration, by using transwell chambers, and angiogenesis, by using in vitro angiogenesis kit. Results Compared with the blank group, the number of EPCs colony, proliferation, adhesion, migration and angiogenesis were all improved in the high-dose group, mid-dose group and simvastatin group （P 〈 0.05）, but had no significant changes in the low-dose group. Conclusion The anti-atherosclerosis effect of Dang Gui Bu Xue Tang is closely related to the promotion of EPCs number and activity.