反义寡核苷酸对人增生性瘢痕成纤维细胞FAK和胶原合成的作用

Effects of antisense oligonucleotides on the expression of focal adhesion kinase gene and collagen sythesis in the cultured human fibroblasts of hypertrophic scar

目的研究黏着斑激酶（focal adhesion kinase，FAK）在人增生性瘢痕发病机制中的作用。方法体外分离、培养人增生性瘢痕成纤维细胞（hyperthrophic sear fibroblasts，HSFB）。在脂质体介导下将FAK反义寡核苷酸转染至HSFB中设为FAK反义寡核苷酸治疗组（AT组），仅加入脂质体为脂质体对照组（LPC组），不加脂质体和反义寡核苷酸为空白对照组（M组）。通过荧光定量PCR方法检测HSFB细胞中FAKmRNA指数，采用^3H-脯氨酸掺入法测定HSFB细胞的胶原合成量。结果转染48h后，AT组FAKmRNA值为0．043±0．030，明显低于LC组（0．124±0．070）及LPC组（0．127±0．0195，P〈0．05）。AT组的^3H-脯氨酸掺入率为257．0±15．14，低于LC组（962．2±300．5）及LPC组（930．8±28．97，P〈0．01）。结论FAK反义寡核苷酸能抑制体外培养的HSFB中FAK基因的表达和胶原合成，FAK在人增生性瘢痕的发病中发挥了一定作用。

Objective To study the role of focal adhesion kinase （FAK） in the pathogenesis of human hyperthrophic scar. Methods Human hyperthrophic scar fibroblasts （HSFB） were isolated from human hypertrophic scar and cultured in vitro. The cells were then divided into 3 groups as AT group（phosphorothioate FAK ASODN was transfected into the HSFB by liposome）, LPC group （liposome only）, and LC group （ control group, without liposome or ASODN）. The FAKmRNA index of HSFB was assessed by polymerase chain reaction method（ FQ-PCR）. The collagen sythesis of HSFB was assessed by 3 H-proline incorporation method. Results The FAK mRNA index of HSFB in AT group 48 hours after trasfection was significantly lower than that in LPC and LC groups（0. 043± 0.030,0. 124 ± 0. 070,0.127± 0.0195, P 〈 0.05）. The 3 H-proline incorporation rate in AT group was lower than that in LPC and LC groups（257.0 ±15.14, 962.2 ± 300.5,930.8 ± 28.97, P 〈 0.01 ）. Conclusion The expression of FAK gene and collagen synthesis of the cultured HSFB could be inhibited by FAK ASODN, indicating that FAK played a rote in the development of excessive fibrosis of human hypertrophic scar.