Study On Ziziphus Jujubes Mill "Jinsixiaozao" For Inhibiting Lung Cells Into Fibrosis Induced By Transforming Growth Factor-β1

In this paper, we focus on both polysaccharide extraction process of Ziziphus JujubesMill”jinsixiaozao” and the effect of chemical constituents for the human alveolar epithelial typeⅡ A549cell line induced into pulmonary fibrosis by Transforming growth factor-β1. This paperproceeds as follow.Section1researched the extraction process of polysaccharides and other ingredients. Theoptimal conditions for the ingredients extraction were defined as75%ethanol,0.5hours,15:1liquid and solid ratio,800W,60℃. And then, this solution processed through petroleum,chloroform, acetic ether and butyl alcohol. The different extractive layers were made into thedry extractum respectively. The Box-Behnken experimental design and the response surfacemethod were used to optimize the polysaccharides’ extraction. Polysaccharide extraction ratewas choosed as observation index. We determine that the best process was microwave extraction.The optimal conditions were defined as3minutes,25:1liquid and solid ratio, and420W. Crudepolysaccharide was degreased by petroleum ether, deproteinized by Sevag method and bleachedby activated carbon. Ultimately, pure polysaccharide was obtained.Section2researched the cytological observation. We found that the cells’ shape changedfrom cobblestoning into fusiformis, and the ligation among the cells became loose after beingtreated with TGF-β1under the inverted microscope for24hours and48hours. Moreover, theshapes changed back to normal cobblestoning or ellipses with TGF-β1and the total composition,polysaccharides, n-butoxide phase and ethyl acetatethe phase, while the cell shape much moretrends to normal with the increasing concentrations of the drugs. In the other way, the cell shapemuch more trends to normal with the lower concentrations of the drugs within TGF-β1and thechloroform phase and the petroleum ether phase, and in the higher concentrations groups, thecells began to shrink even to death.Section3researched the protein expression. The Western blotting method and indirectimmunofluorescence both were used to test the protein expression of the Fibronectin-EDA,which was showed as the mesenchymal cell phenotype. The result showed that the extraction ofZiziphus jujuba could descend the influence of the fibronectin Fn-EDA in the mesenchymal cellstreated by TGF-β1, significantly, while the influence of the fibronectin Fn-EDA in themesenchymal cells treated by TGF-β1was more than the normal. It also showed that the certainconcentration of Ziziphus jujuba’s components had the greatest inhibition. Section4researched the detection of cell proliferative activity. MTT method was chose toresearch the proliferation effects (the absorbance of living cells) of the extracts to A549cellsinduced by TGF-β1for48hours. The results showed that the absorbance value only with TGF-β1was higher than the normal cells, and the absorbance value of the test groups are higher thanthe control group. Compared the control group with experimental groups, the P values hadstatistical significance (P<0.05). As the result, the certain concentration of Ziziphus jujuba’scomponents could inhibit A549into idiopathic pulmonary fibrosis induced by TGF-β1.Our study showed that Ziziphus jujuba’s chemical components could inhibit the humantype II alveolar epithelial cell lines A549into ephethlial-mesenchymal transition (EMT) inducedby TGF-β1in vitro differentiation. Its mechanism was to inhibit its original formation, whichwas ephethlial-mesenchymal transition (EMT). The Ziziphus jujuba had very active role in theprevention or suppression of pulmonary fibrosis process.