Study On The Mechanism Of Emodin And Baicalin On The EMT Of A549 Cells Induced By TGF-β1 Based On Autophagy

Objective:The model of epithelial mesenchymal transition（EMT）in A549 cells induced by transforming growth factor-β1（TGF-β1）was used to screen the active substances of traditional Chinese medicine which can inhibit EMT in vitro.Based on the expression of autophagy related genes and the change of autophagy activity,the potential mechanism of the treatment of idiopathic pulmonary fibrosis（IPF）was explored.Methods:1.Experimental study:the IPF model of EMT in A549 cells induced by TGF-β1was used.baicalin,artesunate,emodin,selaginella extract（alcohol extract,water extract and alcohol precipitate）,matrine,oxymatrine,quercetin,curcumin,humulus alcohol extract（dry and fresh）,rosmarinic acid,ligustrazine,rutin,baicalein,luteolin and liquorice were given respectively q RT-PCR was used to detect the expression of Col I（collagen I）,EMT markers（Vimentin,E-cadherin）and autophagy markers（ATGs）gene m RNA.2.Under inverted phase contrast microscope,we observed the effect of the active substance of Chinese medicine with inhibition of EMT on the morphology of A549 cells induced by TGF-β1;further,we used WB method and autophagy flow to detect the difference of autophagy expression,to explore the mechanism of the active substance of Chinese medicine in inhibiting IPF by autophagy in vitro.result:1.In the model group,the expression of cell type marker gene and microscopic morphological index were consistent with the change of EMT,while the expression of autophagy gene（maker）decreased.The optimal concentration of TGF-β1 for induction of A549 epithelial stromal transformation and reduction of autophagy was 10 ng·ml^-1for48 hours.2.Compared with the model group,after the application of 18 TCM monomers and extracts,the index of Col I decreased to some extent,among which baicalin,artesunate,emodin,humulus alcohol extract,luteolin and quercetin had better inhibitory effect on Col I expression,while the expression of vimentin also changed in the same direction;baicalin and emodin intervened in ATGs of autophagy gene m RNA level was the most significant.3.Morphological observation under inverted phase contrast microscope showed that baicalin inhibited TGF-β1-induced EMT in A549 cells,which made most of the cells maintain epithelial cell morphology.Compared with the model group,the results of q RT-PCR and WB showed that the expression of autophagy related genes ATGs m RNA and autophagy protein was up-regulated in baicalin group and emodin group,and Col I and Vimentin were up-regulated The expression of m RNA was down regulated,and the change range of emodin group was larger than that of baicalin group.The autophagic flow experiment confirmed that the autophagic flow of model group was inhibited,and baicalin and emodin could improve the autophagic activity of cells.Conclusion:Based on the autophagy of EMT,TGF-β1 was used to induce EMT in A549 cells,and q RT PCR was used to detect the expression of Col I,EMT markers and autophagy related genes.In this study,emodin and baicalin can reduce the expression of Col I and vimentin m RNA in A549 induced by TGF-β1,inhibit the EMT process,increase the expression of ATGs m RNA and protein related to autophagy,and improve the autophagy activity of cells...