ERK1/2信号传导通路介导碱性成纤维细胞生长因子诱导人晶状体上皮细胞中环氧合酶2的表达

背景：碱性成纤维细胞生长因子在各种白内障形成中都起着重要的作用，可促进晶状体上皮细胞的增殖并化生为纤维细胞，但其信号通路尚不清楚。目的：探讨ERK1/2信号转导通路在碱性成纤维细胞生长因子诱导的人晶状体上皮细胞环氧合酶2表达中的作用。方法：使用10 μg/L的碱性成纤维细胞生长因子干预培养的人晶状体上皮细胞0，1，3，6，12，24 h，RT-PCR检测刺激不同时间人晶状体上皮细胞环氧合酶2 mRNA的表达，Western blot检测细胞中环氧合酶2及磷酸化ERK1/2的表达。在阻断实验中应用特异性ERK1/2的阻断剂PD98059 阻断ERK信号转导通路1 h，再用10 μg/L碱性成纤维细胞生长因子刺激细胞6 h，Western blot检测细胞中环氧合酶2的表达。结果与结论：碱性成纤维细胞生长因子刺激后的人晶状体上皮细胞中环氧合酶2 mRNA及其蛋白的表达显著增加(P < 0.01)，同时碱性成纤维细胞生长因子诱导人晶状体上皮细胞磷酸化ERK1/2活性增强，表达水平随作用时间而增加，30 min时达到最高峰(P < 0.01)，6 h后恢复至基线水平；PD98059可抑制人晶状体上皮细胞环氧合酶2的表达 (P < 0.01)。说明ERK1/2信号转导通路参与了碱性成纤维细胞生长因子诱导的人晶状体上皮细胞中环氧合酶2的表达，在后发性白内障的形成过程中起着重要作用。

Human lens epithelial cells respond to basic fibroblast growth factor by ERK1/2 regulated induction of cyclooxygenase-2

BACKGROUND:Basic fibroblast growth factor (bFGF) plays an important role in the formation of cataract and promotes the lens epithelium cells proliferation and diffraction to fibroblast cells, but the signal pathway remains poorly understood.  OBJECTIVE:To investigate the role of ERK1/2 signaling pathway in bFGF-induced the expression of cyclooxygenase-2 (COX-2) in human lens epithelial cell line HLB-C3. METHODS:Human lens epithelial cell line HLB-C3 was co-incubated with bFGF (10 μg/L) for 0, 1, 3, 6, 12, and 24 hours. RT-PCR technique and western blot were used to detect the expressions of COX-2mRNA and protein at different times. Specific ERK1/2 inhibitor PD98059 was added into the culture of HLB-C3 cells in the block examination for 1 hour, followed by bFGF   (10 μg/L) stimulation for 6 hours. Western blot technique was used to detect phosphorylated ERK1/2 expression. RESULTS AND CONCLUSION: Theexpression of COX-2mRNA and protein was increased significantly after the bFGF stimulation (P < 0.01); The activity of ERK1/2 was reached a peak at 30 minutes after treatment and decreased to base level at 6 hours (P < 0.01); The expression of COX-2 was down-regulated in the PD98059 group in comparison with that in the bFGF group (P < 0.01). bFGF induces the expression of COX-2 in human lens epithelial cell line. The effects are mediated, at least in part, through the activation of ERK1/2 signaling pathway which plays an important role in the formation of the posterior capsular cataract.