剪切应力作用下麝香保心丸促内皮祖细胞功能的研究  

Promotion of Function of Endothelial Progenitor Cells with Shexiang Baoxin Pill Treatment under Shear Stress

作　　者：李刚[1] 陈旸[1] 吴江[2,3] 

机构地区：[1]四川省人民医院心脏内科,成都610072 [2]四川大学华西医学中心生物医学工程研究室,成都610041 [3]新疆医科大学基础医学院,乌鲁木齐830011

出　　处：《生物医学工程学杂志》2015年第4期847-853,共7页Journal of Biomedical Engineering

基　　金：新疆维吾尔自治区自然科学基金资助项目(201233146-9)

摘　　要：为观察剪切应力作用下麝香保心丸(SBP)对内皮祖细胞(EPCs)功能及其修复动物血管损伤模型的作用,本实验采用密度梯度离心法分离传代培养EPCs,待培养至第三代EPCs细胞贴壁长满后,加入麝香保心丸干预24h,再对其施加15dyne/cm2剪切应力24h,采用CCK-8法、Edu标记法、黏附实验、boyden小室实验及Matrigel法检测EPCs增殖、黏附、迁移及体外微血管形成功能等指标;Western blot法检测EPCs内皮型一氧化氮合酶(eNOS)蛋白表达;同时,建立血管损伤动物模型,分析EPCs细胞移植的血管损伤修复作用。结果显示,与单纯的SBP组比较,剪切应力作用下SBP明显增强了EPCs细胞的增殖能力、细胞迁移率和黏附能力(P值均<0.01),EPCs的微血管形成功能亦显著提高(P<0.01);Western blot条带灰度分析显示eNOS表达水平明显升高(P<0.01);动物模型血管切片的免疫荧光染色结果显示内皮再生率显著增加。可见,剪切应力作用下麝香保心丸能够显著改善EPCs功能,有利于EPCs发挥血管损伤修复作用。The aim of this study was to investigate whether shear stress could promote function of endothelial progen- itor cells （EPCs）with Shexiang Baoxin Pill （SBP） treatment in vitro, and to study whether shear stress contributed to vascular injury repair by EPCs. EPCs were isolated and characterized; EPCs＂ proliferation, migration, adhesion, tube formation and eNOS protein level in vitro were investigated by culturing confluent EPCs in 4 mg/mL SBP under physiological shear stress （15 dyne/cmz） for up to 24 hours. Afterwards, EPCs were transfused into rats after wireinduced carotid artery injury augmented re-endothelialization. The results showed that, compared to the SBP group, the shear stress-l-SBP group obviously enhanced EPCs proliferation, migration, adhesion, tube formation and eNOS protein expression in vitro （P〈0. 01）. After one week, immunofluorescence staining showed that endothelial regeneration rate obviously enhanced in shear stress＋SBP group （P〈0.01）. The present study demonstrates that shear stress can promote function of endothelial progenitor cells treated with SBP, which improves the vascular injury repair potentials of EPCs.

关 键 词：剪切应力 麝香保心丸 内皮祖细胞 再内皮化 

分 类 号：R285[医药卫生—中药学]