Study Of Biotransformation For Liquiritigenin In Licorice With Inhibiting Tumor And Chronic Hepatic Injury Activity

It is important for us to enrich active ingredients from Chinese herbal medicine and develop their new applications that edible and medicinal fungi ferment Chinese herbal medicine. But it also faces many difficults due to complex composition of Chinese herbal medicine and fungi mix, and many factors affect the transformation of Chinese herbal medicine by fungi. Therefore, here is to study the effect and regularity of transformation of medicinal fungi on Chinese herbal medicine using solid transformation system. We set up the solid phase transformation system of licorice, and explore the effect of the biological modification and enrichment on the active fractions of licorice and the anti-tumor mechanism of the active transformed ingredients.Licorice contains a variety of active chemical constituents, such as glycyrrhizic acid and licorice flavonoids, is widely used in the pharmaceutical, food, cosmetics, cigarettes and other industries. In recent years liquiritigenin in licorice flavonoids has important medicinal value, but its separation and purification is tedious and high consumption because of its low concentrations and numerous similar natural derivatives.The main results obtained are as follows:17 kinds of raw materials of Chinese herbal medicine were studied, and the biological pretreatment of 12 kinds of edible and medicinal fungi in Shennongjia Nature Reserve were studied. The suitable strain and transformation model for the effective transformation of Chinese herbal medicine was confirmed. By the effect of different edible and medicinal fungi on the active site modification and enrichment of compounds in licorice, the effect of these different biological properties fungi on the main compounds of licorice root was significantly different. It is reported that the dominant strain of the solid state transformation of licorice was the DS₁.By changing the conditions of the strain DS₁ fermentation, the effect of the biological transformation was optimized. Compared with the control groups, the effect of DS₁ degradation on the main active components of licorice was little. In 14 day the average weight loss was 11.9%. The relative conversion rate was 91.3%, and the yield of liquiritigenin was 5.4 times.Strain DS₁ was identified by the laboratory to determine the taxonomic status: Eumycophyta,Basidiomycetes,Agaricales,Schizophyllaceae,Schizophyllum. It belongs to a kind of edible and medicinal fungi?Schizophyllum commune Fr.?. The results showed that S. commune DS₁ transformed licorice compound with special mechanism.In order to study the main pharmacological effects of biological enrichment, the samples were separated and purified by different treatments, and the substrate and product transformed by DS₁ was identified by the determination of HPLC and NMR.Through analysis of molecular structure liquiritigenin is mainly due to deglycosylation of liquiritin in licorice. Using saturated ammonium sulfate, hydrophobic column and anioncolumn chromatography, Beta glucosidase enzyme from S. commune DS₁ is isolated and pured. Its molecular weight is about 106 KD, the optimum temperature is about 50?, the optimum p H is about 5 6. It was confirmed that the conversion of liquiritin in licorice by beta glucosidase enzyme and crude enzyme of DS₁. is 20.3% and 22.1% respectively. Thus the increase of product liquiritigenin in licorice was enhanced by the transformation of DS₁.The biological function of biotransformation products by DS₁ showed that they can effectively inhibit the activity of human liver cancer cells, down-regulate the expression level of HIF1? and target gene VEGF in SMMC7721. In Hep G2 cells the Beclin-1 gene was up-regulated, the cell cycle gene Cyclin D1 was slightly down regulated, and the biological behavior about apoptosis of Hep G2 cells was regulated. The effect of biotransformation products by DS₁ on the biological function of the hypoxia signaling pathway in the liver cancer cells was indicated.Liquiritigenin is a kind of compound with antioxidant activity, which has important significance for the prevention of solid tumor. In experiment of transformation product liquiritigenin inhibiting chronic exposure of CCl4, the activity of SOD and GSH-Px were increased by 23.5% and 97.1%. SOD2 and GSH-Px m RNA expression level was increased by 0.05 and 102.3%, Rel A expression level was increased by 16.3%, MMP2 were decreased by 50%, Bcl-x were decreased by 66%, and MDA 58%. It was clear that PGC-1? might play a central role in liver injuries leading to cirrhosis and liver cancers. In conclusion, liquiritigenin showed strong efficacy in inhibiting chronic exposure of CCl4 caused hepatic injuries through PGC-1? mediated derepression of antioxidant enzymes, oxidative metabolism genes, and mitochondrial pathway antiopoptotic genes. The results showed that the effective control of the tumor about liquiritigenin from DS₁ mediated transformation of Radix Glycyrrhizae was not seen in the domestic and international. To study the regulation of the potential specific molecular targets for the development of hepatocellular carcinoma provide evidence for the clinical efficacy of the treatment of liquiritigenin.The study revealed the intrinsic mechanism of the transformation in licorice about S. commune DS₁ using HPLC, LC-MSn and NMR analysis to identify the solid state transformation. The anti oxidative mechanism of liquiritigenin, which was from the transformation in licorice by DS₁, through the PGC-1? signaling pathway in the treatment of the liver cancer induced by carbon tetrachloride in mice is also study.