目的：探究吡咯脘二硫代氨基甲酸酯(PDTC)对转化生长因子-beta2(TGF-β2)诱导人晶状体上皮细胞(LECs)上皮-间充质转化(EMT)的逆转机制。方法：TGF-β2诱导人晶状体上皮细胞EMT，并给予不同浓度PDTC处理TGF-β2诱导的LECs。CCK-8、划痕试验检测细胞增殖与迁移，Western Blot检测EMT标志物E-cadherin、α-SMA和NF-κB信号传导通路相关蛋白表达，以及凋亡相关蛋白BAX、BCL-2、Caspase-3、细胞周期蛋白D1的表达。结果：TGF-β2处理组细胞增殖和迁移活力明显高于未添加TGF-β2的实验组； E-cadherin表达下调，α-SMA表达上调。在TGF-β2的作用下NF-κB p65和磷酸化的NF-κB p65表达增加(P<0.05)且在TGF-β2 10 ng/mL刺激浓度时LECs表现出较强的增殖活力和迁移能力。PDTC逆转EMT和诱导细胞凋亡的机制研究表明，PDTC干预处理后细胞活力和迁移能力都显著降低，PDTC抑制IκB的磷酸化进而抑制NF-κB核移位，NF-κB信号通路中NF-κBp65/p-NF-κB p65、Iκκ-α/p-Iκκ-α表达下调(P<0.05)，诱导促凋亡蛋白BAX/Caspase-3表达上调、抑凋亡蛋白BCL-2、细胞周期蛋白Cyclin D1表达下调，NF-κB/IκB mRNA表达下调，凋亡相关mRNA BAX表达上调BCL-2表达下调。结论：PDTC可显著逆转由TGF-β2诱导的LECs细胞EMT过程，可能与抑制NF-κB信号通路活化使NF-κB p65/IκB/Iκκ-α表达降低以及激活凋亡相关蛋白表达有关，PDTC可通过抑制NF-κB信号通路达到逆转EMT的进程并使异常增殖的细胞发生凋亡，这将为后发性白内障的治疗提供新的潜在的治疗药物。

AIM: To investigate the mechanism of pyrrolidine dithiocarbamate(PDTC)on transforming growth factor-beta 2(TGF-β2)-induced epithelial-mesenchymal transition(EMT)in human lens epithelial cells(LECs).METHODS: LECs were treated with various doses of PDTC chemicals following TGF-β2 caused EMT on these cells. Cell proliferation and lateral migration were discovered using the CCK-8 and cell scratch test. The markers of EMT, including E-cadherin, α-SMA and nuclear factor-κB(NF-κB)signaling pathway-related expression, were tested by Western Blot as well as the changes in the expression of the apoptosis-related proteins BAX, BCL-2, Caspase-3, and Cyclin D1.RESULTS: The proliferation and migration viability of cells in the TGF-β2 treated group was increased compared to the group without TGF-β2, and the expression of α-SMA increased whereas the E-cadherin expression decreased. With the effect of TGF-β2, NF-κB p65 and phosphorylated NF-κB p65 expression increased, the concentration of TGF-β2 that had the greatest capacity for proliferation and migration was 10 ng/mL(P<0.05). Mechanism study of PDTC-induced EMT reversal and apoptosis showed that cell viability and migratory capability were both significantly reduced after PDTC intervention; PDTC prevents IκB phosphorylation, thus inhibiting NF-κB nuclear translocation. Protein associated to the NF-κB signaling pathway, and protein expression of NF-κB/IκBα/p-IκBα/Iκκ-α/p-Iκκ-α was decreased(P<0.05), PDTC increased the expression of the pro-apoptotic protein BAX/Caspase-3, expression of the inhibitor of apoptosis protein BCL-2 and the cell cycle protein Cyclin D1 was reduced. The expression of NF-κB/IκB mRNA was reduced, expression of the apoptosis-related mRNA BAX increased, while BCL-2 reduced.CONCLUSION: The EMT in LECs cells induced by TGF-β2 can be significantly reversed by PDTC, which may be related to the decreased expression of NF-κB p65/IκB/Iκκ-α and activation of apoptosis-related protein. PDTC can reverse EMT by inhibiting NF-κB signaling pathway and induce apoptosis of abnormally proliferated cells, which will provide new potential therapeutic agents for posterior capsular opacification(PCO)treatment.

武汉市卫生委员会医学研究项目(No.WX20A15); 湖北科技学院五官医学院专项科研基金项目(No.2020XZ36); 临床研究机构基金项目(No.AR2110D2)