摘要
目的研究转分化过程中晶状体上皮细胞NF-κB信号通路相关蛋白p65,E-cadherin及α-SMA的表达,探讨NF-κB信号通路在晶状体上皮细胞转化中的作用。方法体外培养的HLECs,选择传3代的细胞进行实验,以TGF-β2作为诱导剂。利用NF-κB的特异抑制剂吡咯二硫氨基甲酸(PDTC)进行预处理,采用Trans AM NF-κB p65活性检测试剂盒检测NF-κB p65活性,免疫荧光方法和RT-PCR检测HLECs E-cadherin和α-SMA及其mRNA的表达。结果 TGF-β2处理晶状体上皮细胞后,p65和α-SMA的表达明显增强,细胞E-cadherin表达明显减弱。PDTC能部分逆转HLECsα-SMA和E-cadherin的表达,与对照组比较,差异均有统计学意义。结论 TGF-β2诱导了HLECs转分化,NF-κB信号通路可能参与了TGF-β2诱导的晶状体上皮细胞的转分化过程。
Objective To study the expression of NF-κB p65,E-cadherin andα-SMA in transdifferentiation of human len epithe-lial cells（HLECs） induced by transforming growth factor-β2（TGF-β2） in vitro.To investigate the effect of NF-κB signaling pathway in trans-formation of HLECs .Methods HLECs were cultured and passaged in vitro .The third-generation HLECs were stimulated by TGF-β2 with or without pretreatment of pyrrolidine dithiocarbamate （ PDTC）.Trans AM NF-κB p65 kit was used to detect the activity of NF-κB p65 in HLECs.The expression of E-cadherin and α-SMA in the cells were analysed with immunofluorescent staining .The expression of α-SMA mR-NA and E-cadherin mRNA was detected by RT-PCR.Results The expressions of p65 andα-SMA were increased signifcantly while the ex-pression of E-cadherin was decreased signifcantly after treatment of TGF-β2.PDTC partly reversed the expression of a-SMA and E-cadherin induced by TGF-β2.The differences between the PDTC group and control group were statistically significant （P〈0.05）.Conclusion TGF-β2 induce transdifferentiation of HLECs .NF-κB signaling pathway is probably involved in the process of the transdifferentiation of HLECs in -duced by TGF-β2.
出处
《潍坊医学院学报》
2014年第5期324-326,共3页
Acta Academiae Medicinae Weifang
