维生素C对人胚肺成纤维细胞增殖抑制及凋亡的影响

摘要

目的 研究维生素C对体外培养人胚肺成纤维细胞(human embryo fibroblast,HFL-Ⅰ)增殖、凋亡的影响. 方法 体外培养HFL-Ⅰ细胞,用维生素C处理,分为正常对照组和40、400、4 000 μg/ml维生素C组.于37 ℃继续培养24 h后,用CCK-8法检测细胞增殖情况,用天狼猩红染色法测定细胞内胶原的表达,用流式细胞仪检测细胞周期的变化,用反转录PCR(reverse transcription PCR,RT-PCR)方法检测人胚肺成纤维细胞HFL-Ⅰ中Caspase-3 mRNA的表达差异. 结果 维生素C呈剂量依赖性抑制HFL-Ⅰ细胞的增生(P＜0.05),并降低胶原的生成(P＜0.05),流式细胞仪检测结果显示,细胞主要停滞于G1期,进入S期的细胞明显减少,维生素C作用HFL-Ⅰ细胞12h后Caspase-3 mRNA的表达增强. 结论 维生素C能抑制HFL-Ⅰ细胞的增殖和胶原的增加,其机制可能为作用于细胞周期抑制HFL-Ⅰ细胞分裂及上调Caspase-3的表达诱导HFL-Ⅰ细胞凋亡.%Objective To explore the proliferation and apoptosis of human embryo fibroblast-Ⅰ ( HFL- Ⅰ ) cell lines after cultured with vitamin C. Methods HFL- Ⅰ cell lines were cultured with 0,40,400,4 000 μg/ml vitamin C,respectively. The cell viability was measured by CCK-8 assay. Flow cytometry was used to analyze the cell cycle. Collagen expression was measured by Sirius red staining. The expression of Caspase-3 mRNA was detected by reverse transeription-polymerase chain reaction(RT-PCR). Results Vitamin C inhibited the growth of HFL- I cell lines in a dose-dependent manner(P ＜ 0.05) , and reduced the formation of collagen(P ＜ 0.05). Flow cytometry results showed that the cells were delayed in the G1 phase. Vitamin C significantly up-regulated the mRNA of Caspase-3 after cultured with vitamin C for 12 h. Conclusion Vitamin C can inhibit the proliferation of HFL- Ⅰ , reduce the formation of collagen, block HFL-Ⅰ cells in G1 phase,and induce the apoptosis of HFL-Ⅰ cells by up-regulating the expression of Caspase-3 mRNA.