| Hepatitis B has been epidemic for a considerable long period in all over the world, which is also the most severe and most dangerous killer of all mankind.According to recent research, the infection rate of Hepatitis B in China is around 60%to 70%, and it often attacks children and teenagers. The infection rate of Hepatitis B is on a rise in recent years.Hepatitis B was caused by Hepatitis B virus(HBV), which invades hepatocyte and caused the latter to transform so that leads to reaction in immune system, and thus damage the liver. Hepatitis B has a reputation of long incubation, strong resistance of severe environment, and swift variability. Long time infection of Hepatitis B could cause cirrhosis, hepatic failure, and liver cancer, all of which could seriously influence normal life and even lead to death. Considering Western medical has much negative effects and dependency, people start to look for Chinese traditional medicine curing Hepatitis B.The main purpose of this research was to explore new medicines for anti-HBV therapy, and to obtain the effective ingredients.The subject of this research was natural medicine KY-03, which is used to cure Hepatitis B in rural area of China. KY-03 were extracted by several different solvents in percolation method, and tested with BIAcore SPR to select the effective ones against hepatitis B surface antigen(HBsAg) and hepatitis B e antigen(HBeAg) which are in- dispensable in the life circle of Hepatitis B. Then experiments showed that the crude extracts can inhibit HBV by cell culture experiments in vitro and enzyme-linked immunosorbent assay(ELISA). After the confirmation, using a protein bindling column to purfied the extract and obtain the ingredient named KY-03-B6-4 which against to HBsAg. Analyze the ingredient by HPLC and separate several effective components using Solid-Phase Extraction(SPE) by different densities of solvent.Then compare the results of HPLC-MS analysis with the database of molecular weight and screening a few substances which may have effects on against HBsAg.Employ BIAcore SPR selecting the effective ones called X and Y which both belong to the higher activity of the flavones contents. Lastly, computer molecular docking was adopted to forecast the affinity X to HBeAg by the software Discovery Studio.The experimental results showed that the natural medicine KY-03 has relativelylow toxicity but nice pesticide effect. The ingredient separated from it has more highly efficany, and also the single substances’ efficany possesses superior reliability. |
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