维生素C对人胚肺成纤维细胞增殖抑制及凋亡的影响

Effect of vitamin C on inhibition of proliferation and apoptosis of human embryo fibroblasts

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 000μg/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12 h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-1 （ HFL- Ⅰ ） cell lines after cultured with vitamin C. Methods HFL-Ⅰ cell lines were cultured with 0,40,400,4 000 p,g/ml vitamin C,respeetively. 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 reaetion（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 col-lagen, block HFL-Ⅰ cells in Gl phase, and induce the apoptosis of HFL-Ⅰ cells by up-regulating the expression of Caspase-3 mRNA.