LY294002对bFGF诱导的兔晶状体上皮细胞增生的影响

目的探讨LY294002对碱性成纤维细胞生长因子（bFGF）诱导的兔晶状体上皮细胞（LECs）增生的影响。方法兔眼LECs经培养后，将10^-8、10^-7、10^-6、10^-5、10^-4mol／LLY294002分别加入LECs培养基中培养48h为LY294002组。另将10^-8、10^-7、10^-6、10^-5、10^-4mol／LLY294002＋bFGF（10mg／L）加入培养基中培养LECs48h。bFGF（10mg／L）诱导LECs增生48h为阳性对照组，空白对照组仅用无血清DMEM。MTT比色法测定吸光度（A）值，分析各组药物对LECs增生的抑制率，流式细胞仪检测各细胞周期的LECs百分率。结果bFGF组与空白对照组相比A值升高，LY294002组随着药物浓度的增加，4值明显降低，差异有统计学意义（P〈0．01）。bFGF＋LY294002组与LY294002组相比，A值下降更明显，差异有统计学意义（P〈0．01）。流式细胞仪分析LY294002对LECs的抑制作用与MTT呈现相同趋势，随着LY294002浓度的增加，处于G0／G1期的LECs逐渐增加（P〈0．01），而S期和G2／M期逐渐减少。结论LY294002可明显抑制体外培养的bFGF诱导的兔LECs的增生，并呈剂量依赖关系，为临床筛选防治后发性白内障的药物提供依据。

Effects of LY294002 on bFGF-induced proliferation of lens epithelial cells in rabbit

Background The primary mechanism of posterior capsular opacification （PCO） is the proliferation and migration of lens epithelial cells（LECs） because cytokines stimulate residual LECs following the destroy of blood-aqueous humor. To arrest the proliferation of LECs is very important for the prevention of PCO. Researches showed that LY294002 can inhibit the proliferation of tumor cells. But its effects on LECs is unclear. Objective The aim of this study was to investigate the proliferation inhibition effects of LY294002 on rabbit lens epithelial cells induced by basic fibroblast growth factor, bFGF （bFGF）. Methods The LECs were isolated from rabbits and primarily culture and passaged. 10^-8, 10^-7, 10^-6, 10^-5, 10^-4 mol/L of LY294002 were added into the medium respectively for 48 hours in various experimental groups,and 10^-8,10^-7, 10^-6,10^-5 mol/L,10^-4 mol/L of LY294002 ＋ bFGF（ 10 mg/L） were added for 48 hours in combination drugs groups,and bFGF （ 10 mg/L） was used ior 48 hours to induce the proliferation of LECs in positive control group. LECs was cultured in free-serum DMEM as blank control group. The absorbency（A value） was detected by methyl thiazolyl tetrazolium（MTT） for the assessment of inhibiting rate of LECs in different groups, and percentage of LECs in different cell cycles was tested and analyzed with flow cytometry. Results The A value was elevated in bFGF group in comparison with control group. In 48 hours after LY294002 was added, the A values were gradually and significantly declined with the increase of LY294002 concentration（ P 〈 0.01 ） , and more obvious descend in A values was seen in bFGF ＋ LY294002 groups compared with only LY294002 groups（ P 〈 0.01 ）. The inhibiting rate of LY294002 on LECs showed the same tendency. The percentage of LECs in G0/G1 phase was gradually increased （P 〈0.01 ）, and that in S phase and G2/M phase were significantly decreased with the rise of LY294002 concentration（P 〈 0.01 ）. Conclusion LY294002 can effectively inhibit the proliferation of LECs of rabbit in vitro at a dose-dependent manner. This result offers a basis for the clinical treatment of PCO.