碱性成纤维细胞生长因子在牛晶状体上皮细胞中的表达和调控

Expression and modulation of basic fibroblast growth factor in bovine lens epithelial cells

目的 探讨牛晶状体上皮细胞 (BLEC)中碱性成纤维细胞生长因子 (bFGF)的表达和胎牛血清对表达的调控作用 ,推测白内障摘除术后血 房水屏障破坏对bFGF的影响。方法 采用免疫组化法和Westernblot法检测体外培养的BLEC中bFGF的表达 ,以及不同浓度胎牛血清 (0 1%、1 0 %及 10 0 % )作用 2 4h、10 0 %胎牛血清作用不同时间 (4、8、12及 2 4h)对bFGF相对表达量的影响。结果 免疫组化法检测显示绝大部分BLEC胞浆中可见bFGF表达。Westernblot法检测显示BLEC中表达的bFGF相对分子质量为 180 0 0 ;bFGF的表达随胎牛血清浓度的增加而增强 ,差异有显著意义 (P <0 0 5 ) ;随胎牛血清作用时间的延长而增强 ,差异有显著意义 (P <0 0 5 )。结论 BLEC中可见低相对分子质量bFGF的表达 ;胎牛血清可诱导BLEC中bFGF表达 ;白内障摘除术后血 房水屏障的破坏可导致房水中血清浓度改变 ,引起晶状体上皮细胞中bFGF表达增强。

Objective To evaluate the expression of basic fibroblast growth factor (bFGF) in bovine lens epithelial cells (BLEC), the effects of fetal bovine serum on the expression of bFGF and to explore the effect of breakdown of blood-aqueous barrier on the expression of bFGF after cataract surgery. Methods BLEC were isolated and cultured. Western blot and immunohistochemistry method were used to determine the expression of bFGF in these cells. The dose effects (0.2%, 1.0%, 10.0% fetal bovine serum) and time effects (4, 8, 12, 24 h) of 10.0% fetal bovine serum on the expression of bFGF in BLEC were estimated by Western blot anlysis. Results Immunocytochemical study revealed that the cytoplasm of BLEC could be stained positively by the anti-bFGF antibodies. Western blot analysis revealed the presence of bFGF with a molecular weight of 18 000. The expression of bFGF was upregulated in a dose-dependent and time-dependent manner by fetal bovine serum. There was statistically significant difference among the control group and the fetal bovine serum treated group ( P <0.05). Conclusions BLEC express relatively low molecular weight bFGF. Fetal bovine serum can upregulate the expression of bFGF in the BLEC. Breakdown of blood-aqueous barrier after cataract surgery can stimulate the expression of bFGF in lens epithelial cells through increasing the serum level in the aqueous humor.