摘要: |
目的:通过研究多疣壁虎纤维连接蛋白(fibronectin, FN)在多疣壁虎断尾再生过程中的表达变化及其对Gsn3增殖、迁移和细胞突起生长的影响,初步探讨FN在脊髓再生中的作用及可能机制。
方法:应用简并PCR技术从多疣壁虎组织中获得FN的一段特异性EST序列。采用实时定量聚合酶链式反应(real time quantitative polymerase chain reation, qRT PCR)技术检测壁虎断尾损伤脊髓中FN mRNA的变化。用体外人源FN蛋白培养壁虎少突胶质细胞系Gsn3细胞,DiI染色观察FN对Gsn3细胞形态及增殖的影响;Transwell技术检测FN对Gsn3细胞迁移的影响。
结果:在壁虎断尾损伤后再生脊髓远侧端,FN mRNA表达急剧上调,在断尾后1 d表达最高,随着时间延长逐渐下降。在体外,人源FN促进壁虎Gsn3细胞增殖和迁移,但不能使细胞突起伸长。
结论:FN可能通过促进少突胶质细胞的增殖和迁移参与壁虎断尾后脊髓的再生。 |
关键词: 纤维连接蛋白 再生 Gsn3细胞 增殖 迁移 |
DOI:10.12025/j.issn.1008-6358.2016.20160751 |
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Abstract: |
Objective:To observe expression changes of fibronectin (FN) and its effect on Gsn3 proliferation, migration and oligodendrocyte of spinal cord regeneration of Gekko japonicus, and also analyze the effects and possible mechanism of FN involved in spinal cord regeneration.
Methods:Using degenerate PCR to obtain a special EST sequence of FN from Gekko japonicus tissue. The expression changes of FN mRNA in the regenerating spinal cord after tail amputation were detected by real time quantitative polymerase chain reation (qRT PCR). In vitro, the human FN was used to culture Gekko oligodendrocyte cell line (Gsn3 cells). The morphology and proliferation of Gsn3 cells after FN treatment were also observed by DiI staining, whereas the migration was detected by Transwell.
Results:During the Gekko spinal cord regeneration, FN mRNA expression is significantly increased at 1 dpa (1 day post amputation) then decreased gradually with the time extension in distant spinal cord. FN is able to promote the proliferation and migration of Gsn3 cells, but could not promote the extension of process in vitro.
Conclusions:FN participates in spinal cord regeneration by affecting oligodendrocyte proliferation and migration. |
Key words: fibronectin regeneration Gsn3 cells proliferation migration |