The Effect Of JNK Signaling Pathway On The Fibrosis Of Lens Epithelial Cell

Objective: To study the effect of c-Jun N-terminal kinase(JNK)signal pathway on lens epithelial cells(LECs)fibrosis,and provide theoretical basis for whether JNK can become a potential drug target of posterior capsular opacification(PCO).Methods: The SRA0104 cell line was randomly divided into groups.Cocultured with transforming growth factor-β(TGF-β)and WNT5 a to promote cell fibrosis separately.Adding JNK inhibitor SP600125 to observe the effect of blocking JNK signal pathway on TGF-β and WNT5a-induced LECs fibrosis to explore the role of JNK signaling pathway.The expression of WNT5 a,JNK and p-JNK in each group of cells was detected by Western Blot to analyze the activation of WNT5 a /JNK in cells.Using the same method to detect intracellular collagen I(Col-I),fibronectin(FN),α-Smooth muscle actin(α-SMA)to confirm the effectiveness of WNT /JNK signaling pathway and analyze the active role of JNK signaling pathway in LECs fibrosis.Adding JNK inhibitor SP600125 and detect the protein expression level of SMA Col-Ⅰ,FN and α-SMA.The location and distribution of α-SMA was detected by immunofluorescence method.These methods were used to explore how JNK signal pathway affects the fibrosis of LECs.Transwell chamber test was used to detect the cell migration ability.Col-Ⅰ collagen gel contraction test was used to detect the degree of cell fibrosis,and JNK signal pathway was used to observe the regulation of LECs fibrosis.Results:(1)In SAR0104 cells,TGF-β Stimulate increased expression of WNT5 a,JNK,and p-JNK in cells(P<0.05),accompanied by intracellular Col-Ⅰ,FN,and the protein expression of α-SMA was significantly increased(P<0.05).(2)SRA0104 intracellular mesenchymal cell markers α-SMA exhibits green fluorescence and is widely distributed in the cytoplasm,with a few distributed in the nucleus.After adding JNK inhibitor SP600125,TGF-β JNK,p-JNK,Col-Ⅰ,FN,and the expression of α-SMA was significantly downregulated(P<0.05).(3)Transwell chamber migration experiments showed that TGF-β The number of SRA0104 cells migrating to polycarbonate membrane was significantly increased(P<0.05).(4)The contraction experiment of Col-Ⅰ collagen gel showed that TGF-β Under the action of WNT5 a,the contraction area of Col-Ⅰ significantly decreased compared to the initial area ratio(P<0.05),and SP600125 can significantly inhibit TGF-β The increase in the contraction area of Col-Ⅰ in SRA0104 cells under the action of WNT5a(P<0.05).Conclusion:(1)Fibrosis stimulated by TGF-β in SRA0104 cells is accompanied by activation of the WNT5a/JNK signaling pathway.(2)WNT5a/p-JNK signaling pathway can serve as TGF-β downstream targets of fibrosis.(3)The TGF-β/WNT5a/p-JNK signaling pathway can promote the occurrence of epithelial mesenchymal transition(EMT)in SRA0104 cells,induce extracellular matrix(ECM)deposition,improve cell migration and contractility,and promote the formation of LEC fibrosis.(4)Inhibiting the JNK signaling pathway with SP600125 can effectively block TGF-βstimulated the occurrence of EMT and fibrosis in SRA0104 cells induced by WNT5a/JNK under action.