抑制糖原合酶激酶活性改善动脉粥样硬化小鼠内皮祖细胞功能

Inhibition of glycogen synthase kinase 3βimproves the proliferation,migration and adhesion of endothelial progenitor cells in atherosclerotic mice

目的探讨抑制糖原合酶激酶3β(GSK3β)活性对动脉粥样硬化(As)小鼠内皮祖细胞(EPC)数量、增殖、迁移及黏附功能的影响。方法建立As小鼠模型,密度梯度离心法分离培养As小鼠骨髓源EPC,基因转染法转染抑制GSK3β活性的重组缺陷型腺病毒(基因转染组)至对数生长期的EPC。采用镜下计数法、MTT法、改良Boyden小室迁移试验及黏附试验法测定GSK3β活性对As小鼠EPC数量、增殖、迁移及黏附能力的影响。结果与正常对照组比较,As小鼠EPC数量显著减少,细胞增殖、迁移及黏附能力明显受损(P<0.01,n=5)。与As组比较,抑制GSK3β活性的基因转染组EPC数量(P<0.01,n=5)及增殖能力(P<0.05,n=5)显著增加,EPC的迁移功能及黏附功能均明显改善(P<0.01,n=5)。结论抑制GSK3β活性可增加As小鼠EPC数量并显著改善其增殖、迁移及黏附功能。

Aim To investigate the effect of glycogen synthase kinase 3β(GSK3β)inhibition on the proliferation, migration and adhesion of endothelial progenitor cells(EPC)in atherosclerotic mice. Methods Atherosclerotic mice model were established. Mononuclear cells were isolated and cultured from bone marrow in atherosclerotic mice(atherosclerotic group) and wild type mice(normal control group). EPC in logarithmic phase were transduced with replication defective adenovirus vector expressing catalytically inactive glycogen synthase kinase 3β(GKS3β-KM)(gene transfer group) in atherosclerotic mice. The number and biological functions of EPC, such as proliferation, migration and adhesion were assessed by cells count, MTT assay, modified Boyden’s chamber migrationtrial and adhesion test. Results Compared with normal control group, the number of EPC was observably reduced and proliferation, migration and adhesion capacities of EPC were markedly impaired in atherosclerotic mice(P<0.01,n=5). Compared with atherosclerotic group, the number(P<0.01,n=5)and biological functions of EPC including proliferation(P<0.01,n=5), migration(P<0.01,n=5) and adhesion were significantly enhanced in gene transfer group(P<0.01,n=5). Conclusion GSK3β inhibition could improve impaired proliferation, migration and adhesion capabilities of EPC in atherosclerotic mice.