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ILK介导Akt信号通路诱导人晶状体上皮细胞转分化

摘要 目的 研究ILK抑制剂QLT0267对转分化人晶状体上皮细胞(HLECs)E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、磷酸化Akt(p-Akt)和磷酸化糖原合酶激酶-3β(p-GSK3β)的影响,探讨ILK介导Akt信号通路在HLECs转分化的作用。方法 选择传3代HLECs进行实验,实验分三组:TGF-β2组加入含有浓度为100ng/L的TGF-β2培养基,ILK抑制剂组加入10nmol/LQLT0267预处理1h后,再加入浓度为100ng/LTGF-β2培养基培养48h,对照组加入无血清培养基。Westernblot法检测HLECsE-cadherin、α-SMA、p-Akt和p-GSK3β的变化。采用免疫荧光技术检测HLECsE-cadherin、α-SMA、p-Akt、p-GSK3β的表达。结果 与对照组比较,TGF-β组E-cadherin表达减弱,α-SMA、p-Akt和p-GSK3β的表达增强,差异有统计学意义。与TGF-β组比较,ILK抑制剂组α-SMA、p-Akt和p-GSK3β表达明显减弱,E-cadherin的表达明显增强,差异有统计学意义(F=12.325、17.686、8.547、6.812,P〈0.001),QLT0267部分逆转TGF-β诱导的HLECsα-SMA、p-Akt、p-GSK3β和E-cad-herin的表达。结论 ILK介导Akt信号通路参与TGF-β2诱导的HLECs转分化过程,QLT0267通过特异性结合ILK,影响下游信号的传导,降低了HLECs的转分化。ILK介导Akt信号通路可能在后囊膜混浊过程中发挥重要作用。 Objective To study the effect of QLT0267 on expressions of E-cadherin, α-SMA, p-Akt and p-GSK3β in human lens epithelial cells( HLECs), and investigate the effect of Akt signal pathway mediated by ILK in epithe- lial mesenchymal transition(EMT) of HLECs induced by TGF-β2. Methods The HLECs were cultured and passaged in vitro. The 3rd generation of HLECs were divided into 3 groups. The HLECs in TGF-β2 group were treated with TGF-β2(100ng/L) for inducement for 48 hours. The HLECs in ILK inhibitor group were pretreated withQLT0267 (10 nmol/L) for 1 hour, then treated with TGF-132( 100 ng/L) for 48 h. The HLECs in the control group were cultured in serum-free medium. The expressions of E-eadherin, α-SMA, p-Akt and p-GSK-3β were analyzed by Western blot. The expressions of E-cadherin, α-SMA, p-Akt and p-GSK-3β were detected by cell fluorescence immunoassay. Results Compared with the control group, the expression of E-cadherin was decreased significantly, while the expressions of ot-SMA, p-Akt and p-GSK3β were increased significantly in TGF-β2 group. Compared with TGF-β2 group, the expressions of α-SMA, p-Akt and p-GSK3β were decreased significantly, while the expression of E-cadherin was increased significantly in ILK inhibitor group (F = 12. 325, 17. 686, 8. 547, 6. 812, all P 〈 0. 001 ). QLT0267 partly reversed TGF-β2 induced expressions of E-cadherin,α-SMA, p-Akt and p-GSK3β. Conclusion These results suggest that Akt signal pathway mediated by ILK may be involved in the procedure of EMT of HLECs induced by TGF-β2. QLT0267 can specifically bind ILK, and partly inhibit the process of EMT. Akt signal pathway may play an important role in the progress of PCO.