整合素β1介导的ERK/MAPK通道在人晶状体上皮细胞转分化中的作用  

Effect of integrin β1-mediated ERK/MAPK signal pathway in transdifferentiation of human lens epithelial cells

作　　者：朱艳[1] 朱玉广[1] 张立华[1] 钟莹莹[1] 杜孝楠[1] 张荣[1] 王杰[1] 

机构地区：[1]潍坊医学院附属医院眼科中心,山东省潍坊市261031

出　　处：《眼科新进展》2012年第4期318-321,共4页Recent Advances in Ophthalmology

基　　金：山东省自然科学基金资助(编号:Y2008C127);山东省中青年科学家科研奖励基金资助(编号:2009BSB01638);山东省卫生厅科技计划项目资助(编号:2009HZ105);山东省教育厅科技计划项目资助(编号:J08LG13)~~

摘　　要：目的研究转化生长因子-β2(transforming growth factor β2,TGF-β2)对培养的人氉晶状体上皮细胞(human lens epithelial cells,HLEC)整合素β1、磷酸化ERK(pERK)及F-actin表达的影响,探讨整合素β1介导的ERK/MAPK通路在HLEC转分化中的作用。方法体外培养的HLEC,加入不同浓度(0ng·L-1和10ng·L-1、33ng·L-1、100ng·L-1、333ng·L-1、1000ng·L-1)的TGF-β2处理不同时间(12h、24h、48h、72h),采用MTT法测定TGF-β2对HLEC增殖的影响,并确定最佳干涉浓度和时间用于后续实验。实验分2组,实验组用含最佳干涉浓度TGF-β2的培养基培养HLEC,对照组使用无血清培养基培养,于最佳干涉时间对2组细胞行免疫细胞化学染色检测HLEC中整合素β1、pERK及F-actin的表达,RT-PCR检测整合素β1、pERK及F-actin mRNA的表达。结果 TGF-β2抑制HLEC的增殖,呈浓度和时间依赖性,TGF-β2的最佳干涉终浓度为100ng·L-1,作用48h后达到最大抑制效果。TGF-β2增加HLEC整合素β1、pERK及F-actin的表达,相对表达量分别为0.116±0.031、0.123±0.028、0.140±0.033,均较对照组(0.045±0.011、0.025±0.009、0.079±0.024)明显升高(均为P<0.01)。RT-PCR结果显示,TGF-β2明显促进整合素β1mR-NA、pERK mRNA及F-actin mRNA的表达,相对表达量分别为0.658±0.146、0.582±0.171、0.760±0.193,较对照组(0.246±0.051、0.338±0.092、0.138±0.027)明显升高(均为P<0.01)。结论 TGF-β2抑制了HLEC的增殖,促进整合素β1、pERK及F-actin的表达,整合素β1介导的ERK/MAPK信号通路可能参与了HLEC转分化过程,导致后囊膜混浊。Objective To study the effect of transforming growth factor-β2（TGF-β2） on expression of integrin β1,phosphorylated ERK（pERK） and F-actin in human lens epithelial cells（HLEC）,and investigate the effect of ERK/MAPK signal pathways mediated by integrin β1 in the transdifferentiation of cultured HLEC.Methods HLEC were cultured in vitro and treated with different concentrations（0 ng·L-1,10 ng·L-1,33 ng·L-1,100 ng·L-1,333 ng·L-1,1000 ng·L-1） of TGF-β2 for different time points（12 hours,24 hours,48 hours and 72 hours）.The effect of TGF-β2 on HLEC proliferation was assayed by MTT,and the best time and concentration were determined for further study.The expressions of integrin β1,pERK and F-actin in HLEC were analyzed with immunohistochemistry,and the expressions of integrin β1,pERK and F-actin mRNA were detected by RT-PCR.Results The proliferation of HLEC was suppressed by TGF-β2 in a dose-and time-dependent manner,the maximal inhibition of TGF-β2 on proliferation was at the concentration of 100 ng·L-1 for 48 hours.The expressions of integrin β1,pERK and F-actin after treatment of TGF-β2 were 0.116±0.031,0.123±0.028 and 0.140±0.033,respectively,which in control group were 0.045±0.011,0.025±0.009 and 0.079±0.024,there were statistical differences（both P0.01）.RT-PCR showed that the expressions of integrin β1,pERK and F-actin mRNA after treatment of TGF-β2 were 0.658±0.146,0.582±0.171 and 0.760±0.193,respectively,which in control group were 0.246±0.051,0.338±0.092 and 0.138±0.027,there were statistical differences（both P0.01）.Conclusion TGF-β2 suppresses the proliferation of HLEC and promotes the expression of integrin β1 and pERK.ERK/MAPK signal pathways mediated by integrin β1 may be involved in the transdifferentiation of HLEC,which can result in posterior capsular opacification.

关 键 词：转化生长因子-Β2 整合素Β1 ERK/MAPK通路 F-ACTIN 晶状体上皮细胞 

分 类 号：R692.33[医药卫生—泌尿科学]