碱性成纤维细胞生长因子对体外培养的人晶状体上皮细胞增殖及移行作用的研究
发布时间：2018-07-13 11:18
【摘要】：目的探讨碱性成纤维细胞生长因子(basic fibroblast growth factor, bFGF)对体外培养的人晶状体上皮细胞(human lens epithelial cells ,hLECs)促增殖及促移行作用的影响。初步探讨bFGF对后发性白内障(posterior capsule opacification, PCO)发生、发展中的作用机制,为PCO的防治提供新思路。 方法1.选择人晶状体上皮细胞株(SRA01/04)进行传代培养,观察细胞形态,选择3-5代细胞用于实验。2.在无血清培养液培养的hLECs中分别加入不同浓度的bFGF (0.01、0.10、1.00、10.00及100.00μg/ L),继续分别培养24h、48h、72h后,四甲基偶氮哇盐(4-methyl thiazolyl tetrazolium MTT)比色法测定bFGF促细胞增殖的情况;3.流式细胞术(flow cytometry,FCM)检测细胞周期:选择能稳定促进hLECs增殖作用的bFGF中等浓度10μg/L,分别作用hLECs 24h、48h后流式细胞仪分析细胞周期的改变;4.建立hLECs损伤愈合模型,观察不同终浓度bFGF(0.01、0.10、1.00、10.00及100.00μg/ L),作用hLECs 24h后hLECs的移行情况。 结果1.MTT法显示:bFGF实验组浓度为0.1μg/L、1.00μg/ L、10.0μg/ L、100.00μg/ L时,对hLECs细胞均有明显的促增殖作用,各实验组与阴性对照组比较差异具有统计学意义((P0.05或P0.01),且作用强度呈浓度依赖性,bFGF浓度为100.00μg/ L作用hLECs 24h时,促增殖效果最大(P0.01);2.流式细胞仪分析:实验组bFGF浓度为10μg/L作用24h、48h后,与阴性对照组相比,实验组hLECs细胞周期发生明显改变,表现为G0/G1期细胞显著减少,S期和G2/M期细胞增多,差异有显著性(P0.05),说明bFGF通过促进hLECs细胞越过G1期限止点进入增殖状态; 3.bFGF浓度1.00μg/ L、10.0μg/ L、100.00μg/ L作用hLECs 24h,可明显促进hLECs的移行,其移行能力分别为27.21%(1μg/L)、154.42%(10μg/L)、和238.77%(100μg/L),与阴性对照组相比差异有显著性(P0.01),bFGF对hLECs促移行效应呈剂量依赖关系。 结论bFGF可以促进体外培养的人晶状体上皮细胞(hLECs)的增殖和移行,是hLECs强有力的有丝分裂原和促移行因子,与PCO的形成密切相关。
[Abstract]:Objective to investigate the effects of basic fibroblast growth factor (basic fibroblast growth factor, bFGF) on the proliferation and migration of cultured human lens epithelial cells (human lens epithelial cells / hLECs). To explore the mechanism of bFGF in the occurrence and development of posterior cataract, and to provide a new idea for the prevention and treatment of (posterior capsule opacification,. Method 1. Human lens epithelial cell line (SRA01 / 04) was selected for subculture, cell morphology was observed, and 3-5 passage cells were selected for experiment .2. HLECs cultured in serum-free medium were incubated with different concentrations of bFGF (0.01g / L 0.101.00 渭 g / L) and 100.00 渭 g / L respectively. The proliferation of hLECs was determined by 4-methyl thiazolyl tetrazolium colorimetry. Flow cytometry (flow) was used to detect the cell cycle. The cell cycle was determined by flow cytometry after the medium concentration of bFGF was 10 渭 g / L, which could stably promote the proliferation of hLECs, and the cell cycle was analyzed by flow cytometry for 48 h after 24 h of hLECs treatment. The injury healing model of hLECs was established, and the migration of hLECs was observed after 24 hours of hLECs treated with bFGF (0.01g / L 0.100.100.001.00 and 100.00 渭 g / L). Results 1. MTT assay showed that the concentration of 0.1 渭 g 路L ~ (-1) ~ (-1) 渭 g / L ~ (10. 0 渭 g / L) ~ (10. 0) 渭 g / L ~ (10) 渭 g / L of 10 渭 g / L ~ (10) 渭 g / L ~ (-1) w bFGF could promote the proliferation of hLECs. There was significant difference between the experimental group and the negative control group (P0.05 or P0.01). When the concentration of bFGF was 100.00 渭 g / L for 24 h, the effect of promoting proliferation was the highest (P0.01). Flow cytometry analysis showed that compared with negative control group, the cell cycle of hLECs in the experimental group was significantly changed after the treatment of 10 渭 g / L bFGF for 24 h or 48 h, and the cell cycle in the G _ 0 / G _ 1 phase significantly decreased in S phase and G _ 2 / M phase. The difference was significant (P0.05), which indicated that bFGF could promote the migration of hLECs by promoting the proliferation of hLECs cells over the end of G1 period, while bFGF concentration of 1.00 渭 g / L and 100.00 渭 g / L could significantly promote the migration of hLECs for 24 h. The migration ability was 27.21% (1 渭 g / L), 154.42% (10 渭 g / L) and 238.77% (100 渭 g / L), respectively. Conclusion bFGF can promote the proliferation and migration of cultured human lens epithelial cells (hLECs). BFGF is a powerful mitogen and transporter factor of hLECs and is closely related to the formation of PCO.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R776.1
本文编号：2119172
[Abstract]:Objective to investigate the effects of basic fibroblast growth factor (basic fibroblast growth factor, bFGF) on the proliferation and migration of cultured human lens epithelial cells (human lens epithelial cells / hLECs). To explore the mechanism of bFGF in the occurrence and development of posterior cataract, and to provide a new idea for the prevention and treatment of (posterior capsule opacification,. Method 1. Human lens epithelial cell line (SRA01 / 04) was selected for subculture, cell morphology was observed, and 3-5 passage cells were selected for experiment .2. HLECs cultured in serum-free medium were incubated with different concentrations of bFGF (0.01g / L 0.101.00 渭 g / L) and 100.00 渭 g / L respectively. The proliferation of hLECs was determined by 4-methyl thiazolyl tetrazolium colorimetry. Flow cytometry (flow) was used to detect the cell cycle. The cell cycle was determined by flow cytometry after the medium concentration of bFGF was 10 渭 g / L, which could stably promote the proliferation of hLECs, and the cell cycle was analyzed by flow cytometry for 48 h after 24 h of hLECs treatment. The injury healing model of hLECs was established, and the migration of hLECs was observed after 24 hours of hLECs treated with bFGF (0.01g / L 0.100.100.001.00 and 100.00 渭 g / L). Results 1. MTT assay showed that the concentration of 0.1 渭 g 路L ~ (-1) ~ (-1) 渭 g / L ~ (10. 0 渭 g / L) ~ (10. 0) 渭 g / L ~ (10) 渭 g / L of 10 渭 g / L ~ (10) 渭 g / L ~ (-1) w bFGF could promote the proliferation of hLECs. There was significant difference between the experimental group and the negative control group (P0.05 or P0.01). When the concentration of bFGF was 100.00 渭 g / L for 24 h, the effect of promoting proliferation was the highest (P0.01). Flow cytometry analysis showed that compared with negative control group, the cell cycle of hLECs in the experimental group was significantly changed after the treatment of 10 渭 g / L bFGF for 24 h or 48 h, and the cell cycle in the G _ 0 / G _ 1 phase significantly decreased in S phase and G _ 2 / M phase. The difference was significant (P0.05), which indicated that bFGF could promote the migration of hLECs by promoting the proliferation of hLECs cells over the end of G1 period, while bFGF concentration of 1.00 渭 g / L and 100.00 渭 g / L could significantly promote the migration of hLECs for 24 h. The migration ability was 27.21% (1 渭 g / L), 154.42% (10 渭 g / L) and 238.77% (100 渭 g / L), respectively. Conclusion bFGF can promote the proliferation and migration of cultured human lens epithelial cells (hLECs). BFGF is a powerful mitogen and transporter factor of hLECs and is closely related to the formation of PCO.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R776.1
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