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

摘要

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 scar 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(LC组).通过荧光定量PCR方法 检测HSFB细胞中FAK mRNA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48 h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 flbroblasts (HSFB) were isolated from human hypertrophic scar and cultured in vitro.The cells were then divided into 3 groups as AT group(phosphorethioate FAK ASODN was transfectod into the HSFB by liposome),I.PC group( liposeme 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 rote 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 role in the development of excessive fibrosis of human hypertrophic scar.