碱性成纤维细胞生长因子对人晶体上皮细胞增殖及一氧化氮和诱导型一氧化氮合酶的影响

Effects of basic fibroblast growth factor on proliferation of human lens epithelial cells, nitric oxide and inducible nitric oxide synthase

目的:探讨碱性成纤维细胞生长因子(bFGF)对人晶体上皮细胞内一氧化氮(NO)含量和诱导型一氧化氮合酶(iNOS)活性、基因及蛋白表达的影响。方法:以含10%胎牛血清(FBS)与100mg/L青霉素和100mg/L链霉素的DMEM培养液培养及传代人晶体上皮细胞。用0.001,0.01,0.1,1,10和100μg/L浓度的bFGF作用于晶体上皮细胞,用MTT法检测晶体上皮细胞的增殖情况;用Griess法测定NO含量;分别用RT-PCR方法及westernblot法检测iNOS基因及蛋白的表达。结果:①bFGF对细胞增殖的影响:与对照组比较,除100μg/L组外,其余浓度为0.001～10μg/L的bFGF对人晶体上皮细胞的增殖效应均大于对照组,其中0.01～10μg/L的bFGF对人晶体上皮细胞的增殖效应差异显著(P<0.05)。②bFGF对晶体细胞NO水平、NOS活性及iNOS的基因和蛋白表达的影响:对照组和0.001,0.01,0.1μg/LbFGF组NO水平较低,而在1μg/L和10μg/L的bFGF使人晶体上皮细胞的NO含量增高,0.1～10μg/L的bFGF使人晶体上皮细胞的NOS活性增高、使iNOS的基因和蛋白表达增强。结论:可以促进人晶体上皮细胞的增殖,使NO含量和iNOS的活性和蛋白表达增高。

AIM： To investigate the effects of basic fibroblast growth factor （bFGF） on content of nitric oxide and activity, gene and protein expression of inducible nitric oxide synthase （iNOS） in cultured human lens epithelial cells （HLEC）. METHODS： The HLEC were cultured and passed down by EMEM culture fluid, containing 10% of fetal bovine serum （FBS）, 100 mg/L of penicillin and 100 mg/L of streptomycin. The HLEC was treated by 0.001, 0.01, 0.1, 1, 10 and 100 μg/L of bFGF, and HLEC without intervention of bFGF were taken as control. The MTT assay was used to detect the proliferation of HLEC, and NO content was detected by Griess assay. RT-PCR and western blot were used to detect the protein expression of iNOS. RESULTS： （1）Effects of bFGF on proliferation of cells： the effects of bFGF at all concentrations except 100 μg/L on the proliferation of HLEC were better than the control group, and the differences in effects among b.FGF groups at the concentration from 0.01 μg/L to 10 μg/L were significant （P 〈 0.05）.（2）Effects of bFGF on level of NO, activity of NOS, and gene and protein expression of iNOS： the level of NO in control group and 0.001, 0.01 and 0.1μg/L bFGF group were much lower, while the content of NO in l and 10μg/L bFGF group were increased. The bFGF can enhance the NOS activity and reinforce gene and protein expression of iNOS in HLEC at the concentration of 0.1, 1 and 10 μg/L. CONCLUSION： The bFGF can promote the proliferation of HLEC and enhance the content of NO and the activitym as well as the protein expression of iNOS.