目的 探讨重组Wnt3a蛋白对大鼠血管平滑肌细胞(VSMCs)迁移和黏附的影响及相关机制.方法 原代培养大鼠VSMCs,实验分为Wnt3a组和对照组,通过Transwell实验检测VSMCs的迁移能力,细胞基质黏附实验检测VSMCs黏附细胞外基质的能力,Western blot检测VSMCs中β-连环蛋白(β-catenin)、磷酸化β-catenin(Ser675)、糖原合成激酶3β(GSK-3β),磷酸化GSK-3β(Ser9)、整合素连接激酶(ILK)的蛋白表达水平.结果 Wnt3a组中VSMCs迁移的数量明显高于对照组(P<0.05).与对照组相比,Wnt3a组中VSMCs黏附于胶原Ⅰ的数量和光密度值均显著增加(P<0.05),且Wnt3a组中磷酸化β-cate-nin(Ser675)、磷酸化GSK-3β(Ser9)和ILK蛋白的表达水平均显著上调(P<0.05).结论 Wnt3a可以通过ILK调节VSMCs迁移和黏附.%Objective To investigate the effects and related mechanism of recombinant Wnt 3a protein on vascular smooth muscle cells(VSMCs) migration and adhesion .Methods Primary rat VSMCs were cultured .The experiment was divided into the Wnt3a group and control group .The VSMC migration ability was detected by Transwell experiment .And the ability of VSMC ad-hesion to extracelluar matrix was detected by the celluar matrix adhesion assay .The protein expression levels of β-catenin ,phos-phor-β-catenin (Ser675) ,GSK-3β,phosphor-GSK-3β(Ser9) and integrin-linked kinase(ILK) in VSMCs were detected by Western blot and analyzed .Results The number of VSMCs migration in the Wnt3a group was significantly higher than that in the control group (P<0 .05) .Compared with the control group ,the number of VSMCs adhesion to collagen Ⅰ and the optical density value in the Wnt3a group were significantly increased (P<0 .05) ,and the protein expression levels of β-catenin ,Ser675 ,GSK-3β,Ser9 and ILK in the Wnt3a group were significantly up-regulated (P<0 .05) .Conclusion Wnt3a can regulate VSMCs migration and adhesion by ILK .
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