| Tuberculosis,TB was a major chronic wasting and zoonotic infectious disease caused by Mycobacterium Tuberculosis.Due to uses for decades and irrational uses of anti-TB chemicals in the clinical treatment of tuberculosis,as well as the popularity of AIDS and other reasons,drug-resistant tuberculosis occured.The treatment of drug-resistant TB was difficult and the cure rate was low and will casue the body damage in the process of the treatment.It was imminent to search for new drugs and treatment.Traditional Chinese medicine had a long history to treat disease and less side effects and it was difficult to make the bacteria resistant due to the complex mechanism of action.Therefore,this experiment aimed to search inhibitors or antibacterial synergist of drug-resistant TB from aboudant traditional Chinese medicines and investigate and discuss the mechanism of action preliminarily.Resistance test of Tuberculosis bacterium strains was carried out in this experiment and the results showed that MT-7 strain and MB-1 strain were multidrug-resistant Tubercle bacillus.Therefore MT-7 and MB-1 was choosed as experimental strains,sensitive strain H37 Rv as controls.17 traditional Chinese medicine extract obtained using extraction in water and precipitation in alcohol,respectively,affect on three strains using microdilution method in this experiment.The results showed that three kinds of traditional Chinese medicine,celandine,Radix scutellariae and dandelion all had good antimycobacterial activity in vitro and celandine extract had the best effect.The anti-tuberculosis drugs and three extracts of traditional Chinese medicine affect on jointly H37 Rv,MB-1,MT-1,the results showed that combiantion the celandine alcohol extract with rifampicin had a certain synergy effect to three strains.Total alkaloids in the celandine was extracted and purified.Ttotal alkaloids content in celandine water extract and alcohol extract were deterimined,respectively.Antimycobacterial activity of total alkaloids and water extract and alcohol extract of celandine was compared in vitro by micordilution method.The results showed that the total alkaloids of celandine had the best antimycobacterial activity and alkaloids content or anti-tuberculosis effect in vitro in alcohol extract was higher or better than in water extract.Celandine total alkaloids and anti-tuberculosis drugs affect on jointly three strains,the results showed that the combination had a certain synergy effect.FIC?0.5 indicated that total alkaloids of celandine were antibacterial synergisticagents of rifampicin.In addition,cytotoxicity test of total alkaloids of celandine was detected and MIC value was far below the concentration value of cytotoxicity of the celandine total base.Celandine total alkaloids and combination celandine total alkaloids with rifampicin affect on three strains,respectively.Extraction of RNA of Tuberculosis bacterium was carried out,PhoP and PhoR gene expression quantity were detected by fluorescence quantitative polymerase chain reaction.Test results showed that under the effect of high concentration of celandine total alkaloids,PhoP and PhoR gene expression quantity had decreased in two strains of drug-resistant TB,this indicated celandine total alkaloids can reduce drug-resistant TB.Under the combined effect of celandine total alkaloids and rifampicin,PhoP and PhoR gene expression quantity also had significantly decline in two strains of drug-resistant TB,this indicated that there were antimycobacterial synergist between them and confirmed that celandine total alkaloids was antimycobacterial synergistic agents of rifampicin.The antimycobacterial synergistic mechanism need further research. |
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