堿性成纖維細胞生長因子對體外培養(yǎng)的人晶狀體上皮細胞增殖及移行作用的研究
發(fā)布時間：2018-07-13 11:18
【摘要】：目的探討堿性成纖維細胞生長因子(basic fibroblast growth factor, bFGF)對體外培養(yǎng)的人晶狀體上皮細胞(human lens epithelial cells ,hLECs)促增殖及促移行作用的影響。初步探討bFGF對后發(fā)性白內(nèi)障(posterior capsule opacification, PCO)發(fā)生、發(fā)展中的作用機制,為PCO的防治提供新思路。 方法1.選擇人晶狀體上皮細胞株(SRA01/04)進行傳代培養(yǎng),觀察細胞形態(tài),選擇3-5代細胞用于實驗。2.在無血清培養(yǎng)液培養(yǎng)的hLECs中分別加入不同濃度的bFGF (0.01、0.10、1.00、10.00及100.00μg/ L),繼續(xù)分別培養(yǎng)24h、48h、72h后,四甲基偶氮哇鹽(4-methyl thiazolyl tetrazolium MTT)比色法測定bFGF促細胞增殖的情況;3.流式細胞術(flow cytometry,FCM)檢測細胞周期:選擇能穩(wěn)定促進hLECs增殖作用的bFGF中等濃度10μg/L,分別作用hLECs 24h、48h后流式細胞儀分析細胞周期的改變;4.建立hLECs損傷愈合模型,觀察不同終濃度bFGF(0.01、0.10、1.00、10.00及100.00μg/ L),作用hLECs 24h后hLECs的移行情況。 結果1.MTT法顯示:bFGF實驗組濃度為0.1μg/L、1.00μg/ L、10.0μg/ L、100.00μg/ L時,對hLECs細胞均有明顯的促增殖作用,各實驗組與陰性對照組比較差異具有統(tǒng)計學意義((P0.05或P0.01),且作用強度呈濃度依賴性,bFGF濃度為100.00μg/ L作用hLECs 24h時,促增殖效果最大(P0.01);2.流式細胞儀分析:實驗組bFGF濃度為10μg/L作用24h、48h后,與陰性對照組相比,實驗組hLECs細胞周期發(fā)生明顯改變,表現(xiàn)為G0/G1期細胞顯著減少,S期和G2/M期細胞增多,差異有顯著性(P0.05),說明bFGF通過促進hLECs細胞越過G1期限止點進入增殖狀態(tài); 3.bFGF濃度1.00μg/ L、10.0μg/ L、100.00μg/ L作用hLECs 24h,可明顯促進hLECs的移行,其移行能力分別為27.21%(1μg/L)、154.42%(10μg/L)、和238.77%(100μg/L),與陰性對照組相比差異有顯著性(P0.01),bFGF對hLECs促移行效應呈劑量依賴關系。 結論bFGF可以促進體外培養(yǎng)的人晶狀體上皮細胞(hLECs)的增殖和移行,是hLECs強有力的有絲分裂原和促移行因子,與PCO的形成密切相關。
[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.
【學位授予單位】：寧夏醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R776.1
本文編號：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.
【學位授予單位】：寧夏醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R776.1
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