The Extraction Of Three Chinese Traditional Medicine Ingredient And Preliminary Research On Anti-tuberculosis Role

Objective:In recent years, As drug-resistant tuberculosis bacili heavier and more resistance to the emergence of resistant TB bacillus widespread, and AIDS decline of immunity and human itself, makes tuberculosis is harmful to human health has once again become the number one enemy. At present, the main methods to control TB of human is chemotherapeutic agents. But with the multi-drug-resistant TB cases increased, the gradual losing effectiveness of chemotherapy, the fast-growing drug-resistance of TB, and the long and difficult development cycle of chemical drug, it exceeded its ability. Therefore, it is great significant to search antituberculous Chinese medicine with little side-effect and little drug-resistant. According to reports, Chinese medicine of Radix stemonae,Astragalus and Hop had good curative effects in the treatment of tuberculosis, but its specific medicinal ingredients and the mechanism of action of anti-Mycobacterium tuberculosis were not clear. This experiment extracted these three kinds of Chinese chemical composition by conventional methods, respectively acted on THP-1 cells engulf tuberculosis bacili model, then observed the roles of the three components in the THP-1 cells engulf tuberculosis bacili model and the influence of the three components to THP-1 the cell vitality, which provided the basis for anti-tuberculosi straditional Chinese medicine selection and mechanism research.Methods:In this study traditional methods were used to get three chemical composition of Stemonine,Astragaloside and Lupulone from the Chinese medicine Radix stemonae,Astragalus and Hop. By SephadexLH-20 Gel PW column, the total Stemonine were separated and the monomer Astragaloside were gain. By Reinecke's salt colorimetric method and standard curve method the contents of Stemonine were determined. By Colorimetric method the contents of Astragalosid and Lupulone were determined. The contents of these three kinds of Chinese native medicine were identified through the precipitation reaction, color reaction and TLC. Application THP-1 macrophages model. By THP-1 macrophages model morphologically the action of Stemonine, Astragaloside and Lupulone were observed. By THP-1 macrophages model the whole process of engulf TB and the influence of the three components were observed in the process. By the MTT method Astragaloside and Lupulone's immuno-enhancing effect were validated for THP-1 cells. And By the kit method the change of NO was detected in process of the Lupulone influence on THP-1 macrophage cell engulf TB.Result:By Ethanol Refluxing and Ultrasonic method three chemical compositions of Stemonin, Astragaloside and Lupulone were successfully extracted. By Reinecke's salt colorimetric method the total alkaloid contents which are 0.1 mg/mL were determined. The contents of Astragaloside were 0.7 mg/mL. The contents of Lupulone were 0.1 mg/mL and compared with the extraction rates between the ultrasonic extraction and ethanol refluxing extraction method in the process of the extraction of Lupulone. The extraction rate of the ultrasonic extraction method was higher than ethanol refluxing extraction rate. In the process of applying THP-1 macrophages model, the three drugs were successfully observed, which were impacted on THP-1 cells and the process of macrophage cell engulf TB. By the MTT method the immuno-enhancing of Astragaloside and Lupulone to THP-1 cells were verified, and it obviously increased along with the drug concentration increased. During the detecting process of the Lupulone influence on THP-1 macrophage cell engulf TB to use the kit method, the change of NO showed that the release of NO were increased with Lupulone in the engulf process.Conclusion:Immuno-enhancing for THP-1 cells had been promoted by Astragaloside and Lupulone. the release of NO were promoted by Lupulonein in the whole process of THP-1 macrophage engulf TB. The THP-1 macrophage engulf TB were promoted by Stemonine. There are some respective application value in the treatment process of TB about these three kinds of Chinese medicine.