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

Effect of inhibiting collagen V expression in tenocytes by RNA interference on the fibrillogenesis

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。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。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。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。

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 COLSal and COLSa2 on the mRNA and protein level was significantly inhibited after specific siRNA interference. The inhibition of COLSal and COLSa2 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.