CXCL12/CXCR4对大鼠少突胶质前体细胞迁移的影响及调控

Influence of CXCL12/CXCR4 on the migration of rat oligodendrocyte precursor cells

[目的]研究CXCL12/CXCR4对大鼠少突胶质前体细胞(oligodendrocyte precursor cells,OPCs)迁移的影响及其调控机制。[方法]通过boyden chamber小室检测不同浓度CXCL12(0 ng/ml、5 ng/ml、10 ng/ml、20 ng/ml)对大鼠OPCs迁移能力的影响,利用Western blotting实验检测相应条件下MEK1/2通路中ERK和p-ERK表达的变化;然后利用CXCR4 shRNA抑制CXCR4蛋白表达、U0126阻断MEK1/2通路,利用Western blotting实验检测相应条件下MEK1/2通路中ERK和p-ERK表达的变化,通过boyden chamber小室检测大鼠OPCs迁移能力的变化。[结果]CXCL12能够明显促进大鼠OPCs的迁移,随着胞外CXCL12浓度提高,OPCs内其特异性受体CXCR4表达逐渐增高,与0 ng/ml相比,CXCL12浓度为10 ng/ml和20 ng/ml时CXCR4蛋白相对表达明显增加,差异显著。并且明显促进MEK1/2通路蛋白ERK磷酸水平升高;抑制CXCR4蛋白表达后,CXCL12对少突胶质前体细胞迁移的促进作用受到明显抑制,并且MEK1/2通路蛋白ERK磷酸水平也受到明显抑制;用U0126阻断MEK1/2通路后,CXCL12对大鼠OPCs迁移的促进作用受到明显抑制。[结论]CXCL12/CXCR4能够通过MEK1/2通路调控大鼠OPCs的迁移,为后续深入探讨脊髓损伤治疗方法提供实验基础。

[Objective]To study the effect of CXCL12 / CXCR4 on the migration of rat oligodendrocyte precursor cells（OPCs） and the associated regulatory mechanism.[Method]The effect of CXCL12 on the migration of rat OPCs was assessed by a Boyden chamber assay,and the expression of ERK and p- ERK was detected by western blotting. Next,using CXCR4 siRNA to inhibit CXCR4 expression and U0126 to block the MEK1 /2 pathway,we detected ERK and p- ERK expression by western blotting,and the effect of CXCL12 on the migration of rat OPCs was detected by a Boyden chamber assay. [Result]CXCL12clearly promoted the migration of rat OPCs and increased the phosphorylation level of ERK. After inhibiting CXCR4 expression,the migration of OPCs and the phosphorylation level of ERK were inhibited. The effect of CXCL12 on the migration of rat OPCs was significantly inhibited after the MEK1 /2 signaling pathway had been blocked by U0126. [Conclusion]CXCL12 / CXCR4 may regulate the migration of OPCs via the MEK1 /2 pathway. These results provide an additional foundation for spinal cord injury therapy.