Gualou Xiebai Decoction Inhibited TGF-β1Induced Fibrosis Effect Of A549Cells

The experiment was designed to investigate the role of the extractions from Gualou XiebaiDecoction against idiopathic pulmonary fibrosis. It was induced by the transforming growthfactor TGF-β1working on human an alveolar epithelial type Ⅱcell line A549. The aim was totake advantage of Gualou Xiebai Decoction and understand Gualou Xiebai Decoctionpharmacological efficacy against pulmonary.This thesis included four parts mainly.1. On the basis of single factor experiment, the extraction process of flavonoids in GualouXiebai Decoction was optimized by using response surface methodology, using quercetinextraction rate as index, the best technology of microwave extraction conditions were as follow,1.5min, power of80W, ratio of liquid to solid is50:1. Chemical composition of Gualou XiebaiDecoction was extracted by ultrasonic extraction method, the specific conditions were as follow,the ratio of liquid to solid is50:1,50%ethanol, extraction time is35min and repeat two times.The ultrasonic extraction process was better than that of microwave extraction process. Theextraction was extracted by petroleum ether, chloroform, ethyl acetate and butanol respectively,and then was concentrated. The ethyl acetate phase was separated and purified by silica gelcolumn, using dichloromethane-methanol as mobile phase, and concentrated the solutionobtained from the same proportion of mobile phase to obtain extractum.2. The A549cells were treated with TGF-β1and the total components of Gualou XiebaiDecoction respectively, morphological changes of the cells were observed under invertedmicroscope. The results showed that the morphology of the cells gradually became spindle whenthe A549cells were treated only with TGF-β1. The connections between the cells becameinattentive and the color of cytoplasm became more deeply. After accessing general compositionto intervene on the cell, the morphology of the cells gradually returned to be normal. And thehigher the concentration, the more effects. By using trypan blue staining method for cellcounting, the cell growth curve was drawn.3. MTT Determination of different concentrations on the growth of A549cell lines wasused to test the drug on the inhibition rate of A549cells. The OD values of test groups werehigher than the control groups. The results showed that Gualou Xiebai Decoction could inhibitA549cell to fibrosis.4. Western blotting method could show the expression of the protein by electrophoresis, transmembrane and antibody incubation process. And then the information was obtainedthrough the particular protein expression by the position and the amount. So the total protein ofthe blank groups, the control groups and the test groups were extracted and collected. The resultshowed that Gualou Xiebai Decoction inhibited the expression of A549cells Fn-EDA proteininduced by TGF-β1.Our study showed that Gualou Xiebai Decoction could inhibit A549cells fibrosis. Themechanisms of action and prescription basis were unclear currently, pending further study.