YUAN Jishan 1 , LI Tianzeng 2 , QI Shaohai 2
  • 1Department of Orthopedics, the Affiliated People’s Hospital of Jiangsu University, Zhenjiang Jiangsu, 212002, P.R.China;;
  • 2Department of Burns, the First Affiliated Hospital, Zhongshan University;
Col I A1; Antisense ol igodeoxyneucleotide; Hypertrophic scar; Collagen
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To investigate the inhibitory effect of Col I A1 antisense ol igodeoxyneucleotide (ASODN) transfection mediated by cationic l iposome on Col I A1 expression in human hypertrophic scar fibroblasts. Methods Scar tissue was obtained from volunteer donor. Human hypertrophic scar fibroblasts were cultured by tissue block method. The cells at passage 4 were seeded in a 6 well cell culture plate at 32.25 × 104 cells/well, and then divided into 4 groups: group A, l iposome
and Col I A1 ASODN; group B, Col I A1 ASODN; group C, l iposome; group D, blank control. At 8 hours, 1, 2, 3 and 4 days after transfection, total RNA of the cells were extracted, the expression level of Col I A1 mRNA was detected by RT-PCR, the Col I A1 protein in ECM was extracted by pepsin-digestion method, its concentration was detected by ELISA method. Results Agarose gel electrophoresis detection of ampl ified products showed clear bands without occurrence of indistinct band, obvious primer dimmer and tailing phenomenon. Relative expression level of Col I A1 mRNA: at 8 hours after transfection, group A was less than groups B, C and D (P  lt; 0.05), and groups B and C were less than group D (P  lt; 0.05), and no significant difference was evident between group B and group C (P gt; 0.05); at 1 day after transfection, groups A and B were less than groups C and D (P  lt; 0.05), and there was no significant difference between group A and group B, and between group C and group D (P  gt; 0.05 ); at 2 days after transfection, there were significant differences among four groups (P  lt; 0.05); at 3 and 4 days after transfection, group A was less than groups B, C and D (P  lt; 0.05), group B was less than groups C and D (P  lt; 0.05), and no significant difference was evident between group C and group D (P  gt; 0.05). Concentration of Col I protein: at 8 hours after transfection, group A was less than groups B, C and D (P  lt; 0.05), groups B and C were less than group D (P  lt; 0.05), and no significant difference was evident between group B and group C (P  gt; 0.05); at 1 day after transfection, significant differences were evident among four groups (P  lt; 0.05); at 2, 3 and 4 days after tranfection, groups A and B were less than groups C and D (P  lt; 0.05), and no significant difference was evident between group A and group B (P  gt; 0.05). Conclusion Col I A1 ASODN can inhibit mRNA and protein expression level of Col I A1. Cationic l iposome, as the carrier, can enhance the inhibition by facil itating the entry of ASODN into cells and introducing ASODN into cell nucleus.

Citation： YUAN Jishan,LI Tianzeng,QI Shaohai. INHIBITORY EFFECT OF HUMAN COL I A1 ANTISENSE OLIGODEOXYNEUCLEOTIDE ON COLLAGEN SYNTHESIS IN HYPERTROPHIC SCAR FIBROBLASTS/. Chinese Journal of Reparative and Reconstructive Surgery, 2009, 23(6): 713-717. doi: Copy