To investigate the mechanism by which modified Jiawei Fangji Huangqi Decoction intervenes in the lncRNA NKILA-mediated RIF model of HK-2 cells to inhibit EMT
[Objective]To explore the intervention mechanism of the modified Fangji Huangqi Decoction on the epithelial-mesenchymal transformation in the HK-2 cell fibrosis model mediated by lncRNA NKILA.[Methods]Modified Fangji Huangqi Decoction was applied in an HK-2 cell RIF model built by lncRNA NKILA overexpression lentivirus transfection,with an AG490 control group.qPCR,Western Blot,and immunofluorescence were utilized to assess the alterations in lncRNA NKILA expression,EMT markers FN,Col1,Vim,α-SMA,and JAK2/STAT3 pathway indicators.[Results]EMT phenotypic index analysis indicated up-regulated mRNA and protein levels of FN,Col1,Vim,and α-SMA in the oe-NKILA group versus the oe-NC group,with a noTab.down-regulation in E-cad mRNA and protein levels.Compared to the oe-NKILA group,the oe-NKILA+AG490 and oe-NKILA+TCM-H groups exhibited significant down-regulation in E-cad mRNA and protein levels,with statistical significance(P＜0.05).The expression of lncRNA NKILA in each group was also examined.Compared to the oe-NC group,NKILA expression was significantly elevated in the oe-NKILA group.However,NKILA expression was significantly reduced in the oe-NKILA+AG490 group(P＜0.01)and the oe-NKILA+TCM-H group(P＜0.05).The JAK2/STAT3 pathway indicator analysis revealed significant up-regulation in mRNA and protein,p-JAK2,and p-STAT3 protein expressions of JAK2 and STAT3 in the oe-NKILA group versus the oe-NC group,with significant down-regulation in the oe-NKILA+AG490 and oe-NKILA+TCM-H groups(P＜0.05).[Conclusion]The high-dose serum of the modified Fangji Huangqi Decoction can alleviate the EMT injury of HK-2 cells induced by lncRNA NKILA overexpression and inhibit the activation of the JAK2/STAT3 signaling pathway during the process of lncRNA NKILA-inducing EMT of HK-2 cells.
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