聚乙二醇400对体外培养人角膜上皮细胞生长的影响
The effects of polyethylene glycol 400 on corneal epithelial cell growth in vitro
孙正太 1张晓峰 1夏蔚 1钟蕾 1王英明 1夏静1
作者信息
- 1. 苏州大学附属第一医院眼科,江苏苏州215006
- 折叠
摘要
目的 研究聚乙二醇400对体外培养人角膜上皮细胞( CECs)生物学特性的影响及其分子机制.方法 制备含0.1％聚乙二醇400的DMEM/F12培养基.采用含0.1％聚乙二醇400培养基培养人CECs,倒置显微镜下观察并照相记录细胞培养24、48、72 h时的形态；四甲基偶氮唑蓝(MTT)法检测角膜上皮细胞生长曲线；流式细胞术检测角膜上皮细胞细胞周期；RT-PCR检测角膜上皮细胞中碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)基因表达水平.结果 0.1％聚乙二醇400对角膜上皮细胞的细胞形态无影响,实验组及对照组细胞形态均呈典型的多角形.MTT及流式细胞术检测结果显示,0.1％聚乙二醇400对体外培养人CECs的生长有促进作用(均P＜0.01).0.1％聚乙二醇400培养人CECs后bFGF和EGF的基因表达水平比对照组高(P＜0.05或＜0.01).结论 0.1％聚乙二醇400可能通过缩短人CECs的细胞周期和上调人CECs bFGF、EGF基因的表达,从而加速CECs增殖.
Abstract
Objective To investigate the effects of polyethylene glycol 400 on the biological characteristics of corneal epithelial cells ( CECs) and molecular mechanisms thereof. Methods Polyethylene glycol 400 was diluted to 0-1% with DMEM/F12 medium. CECs were cultured in this medium and cell morphology was recorded using an inverted optical microscope after 24, 48 and 72 hours. Cell growth curves were measured by using MTT assay and cell cycle was determined by using flow cytometer; RT-PCR was used to measure the expression of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in CECs. Results 0.1% polyethylene glycol 400 had no discernable effects in the morphology of CECs. CECs cultured in the dilution of polyethylene glycol 400 solutions have the same polygon cell morphology as the control group. Results from MTT assay and cell cycle measurements showed that 0.1% polyethylene glycol 400 promotes in vitro HCEC growth (P < 0. 01) and up-regulates the expression of bFGF and EGF in cultured CECs( P <0.05 or 0.01). Conclusion 0.1% polyethylene glycol 400 does not affect cell morphology and it promotes CEC proliferation via reducing cell cycle and increasing bFGF and EGF expression.
关键词
聚乙二醇400/人角膜上皮细胞/碱性成纤维细胞生长因子/表皮生长因子Key words
polyethylene glycol 400/human corneal epithelial cells/basic fibroblast growth factor/epidermal growth factor引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据