RNA干扰抑制大鼠肌腱细胞中V型胶原基因的表达

目的：利用RNA干扰技术（RNAi）下调肌腱细胞中V型胶原蛋白的表达,拟为后续探讨V型胶原在损伤肌腱修复过程中的作用提供可行的试验方法。方法：分别设计干扰大鼠V型胶原[COL5（1）2（2）]两个亚基COL5 1和COL5 2表达的siRNA序列,转染大鼠肌腱细胞。通过实时荧光定量PCR和免疫荧光法检测V型胶原在基因和蛋白水平的表达。结果：转染特定的siRNA后,V型胶原两个亚基的基因表达被抑制约70%,蛋白表达也被明显抑制。结论：RNAi法可有效地抑制大鼠肌腱细胞中V型胶原的基因和蛋白水平表达。为进一步研究V型胶原在肌腱损伤修复过程中的作用提供了可行的试验方法。

Inhibition of Gene Expression of Type V Collagen in Rat Tendon Cells by RNA Interference

Objective： Down regulate type V collagen expression in tendon cells using RNA interference （ RNAi ） to provide a viable test method for investigating the role of type V collagen in the injured tendon repair process. Method： Two target siRNA sequences for mRNA of the al and a2 chains in rat type V collagen were designed and transfected into rat tendon cells. Real time PCR and immunofluorescence were used to detect type V collagen expression in gene and protein level. Result： The results showed that the interference effect of the mRNA expression was about 70 % and protein level was siguificantly inhibited after specific siRNA interfection. Conclusion： RNAi method can effectively inhibit the gene and protein expression of type V collagen in rat tendon cells. This study provides a viable test method to further study the role of type V collage in injured tendon repair process.