bFGF与ERK1／2阻断剂对人晶状体上皮细胞α平滑肌肌动蛋白mRNA表达的影响

Effects of bFGF and ERK1/2 blocking agent on α-SMA mRNA expression in human lens epithelial cells

目的观察碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)及细胞外调节蛋白激酶1/2(extracellular signal regulated kinase 1/2,ERK1/2)阻断剂PD98059对人晶状体上皮细胞(human lens epithelial cells,HLEC)α-平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)mRNA表达的影响。方法体外传代及培养HLEC,加入10μg·L-1bFGF及PD98059作用一定时间并根据作用时间进行分组。MTT法检测HLEC数量及存活率;逆转录聚合酶链反应(RT-PCR)测定HLEC中α-SMA mRNA的表达。结果 HLEC的存活率在bFGF作用组中随时间增长而增强,作用6 h时达126.34%,作用1 h组与作用6 h组比较差异有统计学意义(P<0.05);只加PD98059组细胞存活率仅30.03%,与正常对照组相比差异有统计学意义(P<0.05)。bFGF作用剂量不变的情况下,随着作用时间延长,α-SMA mRNA表达明显增加,作用1 h组与作用6 h组差异有统计学意义(P<0.05);bFGF作用6 h后,α-SMA mRNA值达最高;加阻断剂PD98059作用后,α-SMA mRNA表达减少且随阻断剂作用时间延长表达进一步减少,作用1 h组与作用6 h组差异无统计学意义(P>0.05)。结论 bFGF对HLEC具有促增殖作用,在低浓度剂量不变的前提下,呈现时间相关性。bFGF可促进HLEC分泌α-SMA,促进HLEC分化,并呈现时间及剂量相关性。

Objective To observe the effects of basic fibroblast growth factor （bFGF） and extracellular regulated kinase 1/2 （ERK1/2）blocking agent PD98059 on al- pha smooth muscle actin （α-SMA）mRNA expression in human lens epithelial cells （HLEC）. Methods HLEC were primarily cultured and passaged in vitro. The bFGF （ 10 μg. L-1 ）and PD08059 were added and activated for a certain period of time, and then divided according to the activated time. The number and activity of HLEC were de- tected by MTT, and RT-PCR was used to detect the expression of c^-SMA mRNA in HLEC. Results The survival rate of HLEC in bFGF group increased with the time prolong, up to 125.34% at 6 hours, the difference was statistically significant between1 hour and 5 hours （P 〈 0.05 ） ; The cell survival rate in PD98059 group was only 30.03% , there was statistically significant difference compared with the control group （P 〈 0.05 ）. When the dose of bFGF unchanged, as the prolong of time, （α-SMA mRNA expression increased significantly, the difference was statistically significant between 1 hour and 5 hours （ P 〈 0.05 ） ; After bFGF activation for 5 hours, α-SMA mRNA value reached the highest level, after added PD98059, α-SMA mRNA ex- pression reduced, and further decreased with the antagonist action time increased, there was no significant difference be- tween 1 hour and 5 hours （P 〉 0.05 ）. Conclusion bFGF can promote the proliferation of HLEC with time dependence under the stable low dosage, bFGF can promote theα-SMA secretion in HLEC and HLEC differentiation with time and dose dependence.