RNA干扰抑制肌腱细胞V型胶原表达对胶原纤维形成的影响

摘要

Objective:To investigate the role of collagen V in the injured tendon repair by down regulating gene expression in tenocytes using RNA interference (RNAi).Methods:Two target siRNA sequences for mRNA of the 1 and 2 chains in collagen V were designed and transfected into rat tenocytes.The expression of collagen V on the gene and protein levels was detected.A tissue engineered tendon was used to detect the content and fibril diameter of the collagen.Results:The expression of COL5α1 and COL5α2 on the mRNA and protein level was significantly inhibited after specific siRNA interference.The inhibition of COL5α1 and COL5α2 had different effects on fibrillogenesis.Conclusion:This study explores the effecs of collagen V on fibrillogenesis,which may provide a basis for future development of novel cellular and molecular biology-based therapy for tendon injuries.%目的:利用RNA干扰技术(RNAi)下调肌腱细胞中V型胶原基因的表达,探讨V型胶原在损伤肌腱修复过程中的作用.方法:分别设计干扰大鼠V型胶原2个亚基COL5α1和COL5α2表达的siRNA序列,转染大鼠肌腱细胞.检测V型胶原在基因和蛋白水平的表达,检测组织工程肌腱的胶原含量和胶原原纤维直径.结果:转染特定的siRNA后,V型胶原2个亚基的基因和蛋白表达被明显抑制.抑制COL5α1和COL5α2亚基对胶原纤维形成的影响不同.结论:本实验探究了V型胶原对胶原纤维生长自聚的影响,为肌腱损伤修复提供有用的基础生物学信息.