Study On Alizarin Combined With Standard Anti-tuberculosis Chemical Therapy In Treatment Of Multidrug-resistant Pulmonary Tuberculosis

Objective To investigatethe related risk factors between different epidemiological characteristics and drug resistance.To observing the in vitro antimicrobial activity of alizarin against multidrug-resistant Mycobacterium tuberculosis and how alizarin affected non-alcoholic steatohepatitis induced by rifampicin in mouse.To evaluate the clinical efficacy and adverse drug reactions of alizarin combined with anti-tuberculosis therapy for multidrug-resistant pulmonary tuberculosis(MDR-PTB).Methods 63 clinical isolates of mycobacterium tuberculosis which were collected from Hangzhou city were identified with multi PCR Kit method,anayze the susceptibility results and genotyping results.Use gene sequencing method to sequence inhA gene,rpoB gene,rpsl gene,rrs gene,pncA gene of resistant strains,then detect the presence or absence of drug resistance gene mutation and determine the type of mutation.Standard human strains of mycobacterium tuberculosis(H37RV)and multi-drug resistant TB strains were inoculated in different concentrations of alizarin drug culture medium to observe the growth of mycobacterium tuberculosis and evaluate the antibacterial activity in vitro.60 KM mice were divided into 4 groups:normal group(control),alizarin(QCS)group,rifampicin(RFP)group,alizarin and rifampicin combination group(MIX).Mice were sacrificed at each time point.Blood and liver were collected to test liver parameters,liver index,liver pathology,the concentration of malondialdehyde(MDA),the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-PX)in mitochondrion of hepatocytes.A total of 200 MDR-PTB patients received standard anti-tuberculosis treatment for 8 months,and additional alizarin was given to study group.Chi-square test was used to assess the differences in clinical efficacy between two groups(including patients categorized according to TCM syndrome).Result There are 63 drug-resistant strains in 258 clinical isolates of Mycobacterium tuberculosis,accounted for 24.42%.The rate of gene mutation was 50.79%by DNA sequencing,the strains was detected with mutation located in inhA?rpoB?rpsl?rrs gene.Alizarin in 80 pg/mL was showing multi-drug-resistant strains of clinical suppression of better killing effect,compared with positive group,at each time point was indicative of statistical significance(P<0.05).HE staining showed apparent steatosis and inflammation infiltration in RFP group,While steatosis and inflammation were weaker in alizarin group.Serum ALT?AST??-GGT?TBIL?RBIL in RFP group increased significantly comparing to alizarin group and MIX group(P<0.01).While the level of MDA is lower in alizarin group and rMIX group than RFP group.The activity of SOD and GSH-PX expression were downregulated dramatically at week 1,2,4 in RFP group,this trend diminished in MIX group(P<0.05).The clinical effect(39%)and sputum negative conversion rate(76%)(treatment at 8 months)of alizarin combination group were superior to those of single group.For patients with TCM syndrome differentiation as phlegm-heat stagnating lung and those with qi-stagnation induced blood-stasis,alizarin combination therapy had significantly higher total effective rate than standard.anti-tuberculosis treatment(P<0.05).But the difference was not observed in patients with TCM syndrome differentiation as deficiency of qi and blood(P>0.05).The adverse drug reactions rate in study group was 27%,which was significantly lower than that in control group(66%).Conclusion Alizarin in the treatment of patients with MDR-PTB patients can improve the curative effect of chemotherapeutics.