雷帕霉素对体外培养瘢痕疙瘩成纤维细胞增殖的影响

目的:雷帕霉素(Rapamycin,Rapa)对体外培养瘢痕疙瘩成纤维细胞的抑制作用,深入探讨其作用机制。方法:用雷帕霉素的不同药物浓度干预体外培养瘢痕疙瘩成纤维细胞,CCK-8方法检测对瘢痕疙瘩成纤维细胞增殖的影响;免疫组化、RT-PCR和western blot检测干预前后瘢痕疙瘩成纤维细胞中磷酸化的P70s6k、4E-BP1表达情况。结果:CCK-8检测体外培养瘢痕疙瘩成纤维的吸光度,随浓度和时间的增大而减小,各组之间差别具有统计学意义(P0.05);免疫组化(药物浓度取10nmol/L),RT-PCR和western blot对不同药物浓度(0.1nmol/L、1nmol/L、10nmol/L、100nmol/L)干预前后,基因表达和蛋白表达均下降,各组之间差别具有统计学意义(P0.05)。结论:雷帕霉素对体外培养瘢痕疙瘩成纤维细胞的增殖有明显的抑制作用,也抑制瘢痕疙瘩成纤维细胞中磷酸化的P70s6k、4E-BP1表达。

The effect of rapamycin on proliferation of Human Keloids Fibroblasts in vitro

Objective To investigate the effect of rapamycin on proliferation of Human keloid fibroblasts in vitro,to further explore the mechanism of it.Methods With different concentrations of rapamycin effect on cultured keloid fibroblasts,and CCK-8 assay was adopted to evaluate cell survival.lmmunohistochemistry,RT-PCR and western blot were performed to detect the expression of P70s6k and 4E-BP1 before and after intervention.Results CCK-8 assay detected the absorbance of keloid fibroblasts,which decreased with the concerntrations and time increased.There were significant differences between these groups （P＜0.05）.IHC （rapamycin concentration 10nmol/L intervention）,RT-PCR and western blot results showed that gene expression and protein expression of P7Os6k and 4E-BP1 were all decreased in （0.1nmol/L,1nmol/L,10nmol/L,100 nmol/L）rapamycin concentration before and after intervention.There were significant differences between these groups （P ＜0.05）.Conclusions rapamycin significantly inhibited the proliferation of Human Keloids Fibroblasts cell in vitro.rapamycin also inhibited the expression of P70s6k and 4E-BP1.