整合素连接激酶对含成纤维细胞胶原网格收缩的影响

目的 探讨整合素连接激酶(ILK)对含成纤维细胞胶原网格(FPCL)收缩的影响和机制.方法 (1)运用人皮肤成纤维细胞(Fb)构建FPCL,观察ILK-AKT信号通路特异性抑制剂LY294002对FPCL收缩的影响.(2)运用Western blot印迹法检测ILK小干扰RNA(siRNA)转染和LY294002对Fb中ILK和α-平滑肌肌动蛋白(α-SMA)表达的影响.结果 第24小时和第48小时LY294002组FPCL收缩率(20.23±9.57)％、(22.47±10.93)％]明显低于对照组(48.73±6.60)％、( 54.33±12.88)％](P＜0.05),第72、96和120小时FPCL收缩率为(25.58±7.40)％、(26.67±13.76)％、(28.82±3.48)％明显低于对照组(58.00 ±7.54)％、(59.67±11.29)％、(66.95±9.98)％和DMSO组(47.34±12.41)％、(52.26±10.90)％、(56.38±10.75)％ (P＜0.05).LY294002组和ILK siRNA转染组α-SMA蛋白表达(0.992 ±0.255、1.225 ±0.323)与各对照组(2.009 ±0.820、2.190 ±0.577、1.758±0.732)比较均显著降低(P＜0.05).结论 阻断ILK信号通路可明显抑制Fb构建的FPCL收缩和Fb中α-SMA的表达.

Effect of integrin-linked kinase on fibroblast populated collagen lattice contraction

Objective To investigate the influences of integrin-linked kinase (ILK) on fibroblast populated collagen lattice (FPCL) contraction and the mechanism.Methods The human skin fibroblasts from plastic patients were used to construct FPCLs.The effects of ILK-AKT inhibitor LY294002 on FPCL contraction were observed.The expressions of ILK and α-SMA in fibroblasts was detected by using Western blotting after ILK siRNA transfection or LY294002 treatment.Results FPCLs in each group had various extents of contraction,but collagen gels without fibroblasts had no contraction.The contraction rate of FPCLs in LY294002 group at 24 and 48 h ( 20.23 ± 9.57 ) ％,(22.47 ± 10.93 ) ％ ] was significantly lower than that in control group (48.73 ±6.60)％,(54.33 ± 12.88)％ ] (P ＜0.05).It was also significantly lower than that in control group and DMSO group at 72,96 and 120 h (P ＜0.05).The expression of α-SMA in ILK siRNA transfected group (0.992 ±0.255) and LY294002 group ( 1.225 ±0.323)was significantly lower than other groups (2.009 ± 0.820,2.190 ± 0.577,1.758 ± 0.732 ) ( P ＜ 0.05 ).Conclusion The contraction of FPCL and expression of α-SMA in skin fibroblasts can be inhibited via blocking ILK pathway.