CAJ | 학술논문

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COL51和COL52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。
목적：이용RNA간우기술（RNAi）하조기건세포중V형효원단백적표체，의위후속탐토V형효원재손상기건수복과정중적작용제공가행적시험방법。방법：분별설계간우대서V형효원[COL5（1）2（2）]량개아기COL51화COL52표체적siRNA서렬，전염대서기건세포。통과실시형광정량PCR화면역형광법검측V형효원재기인화단백수평적표체。결과：전염특정적siRNA후，V형효원량개아기적기인표체피억제약70%，단백표체야피명현억제。결론：RNAi법가유효지억제대서기건세포중V형효원적기인화단백수평표체。위진일보연구V형효원재기건손상수복과정중적작용제공료가행적시험방법。
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 a1 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 significantly 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.