Pharmacodynamic Study On Chinese Medicine Compound(NBXH) Extracts And Its Combination With Chemotherapy

China is one of 22 high TB burden countries in the world. We studied the antibacterial activity in vivo and in vitro of traditional Chinese medicine compound NBXH extracts prepared by different processes, to provide experimental basis for the preparation of the Chinese patent medicine.1. The bacteriostatic action of Chinese medicine compound NBXH extracts to MDR-M.tb and NTM in vitroPreparare respectively the Lowenstein-Jensen media containing 50,100, 200 mg.mL-1 NBXH water extract and alcohol extract, and preparare respectively the Lowenstein-Jensen media containing 1, 10 ug.ml-1 of isoniazide(INH) and 50, 250 ug.mL-1 of rifampicin(RFP) as the controls. 5multidrug-resistant Mycobacterium tuberculosis(MDR-M.tb) clinical isolates, Mycobacterium bovis and 13 non-tuberculous mycobateria(NTM)standard strains were inoculated respectively on the Lowenstein-Jensen media mentioned above, and then incubated at 37 ℃ for 4 weeks to determine the minimum inhibitory concentrations(MICs). The results showed that the MICs of NBXH water extract on 5 MDR-M.tb isolates(No.13424, No.13420, No.13449, No.13451 and No.13445),Mycobacterium bovis, Mycobacterium intracellulare, Mycobacterium scrofulaceum, Mycobacterium vaccae, Mycobacterium fortuitum strains were 200 mg.mL-1, on Mycobacterium phlei, Mycobacterium smegmatis,Mycobacterium gordonae, Mycobacterium kansasii, Mycobacterium gastri strains were 100 mg.mL-1, and on Mycobacterium avium were 150 mg.mL-1;the MICs of NBXH ethanol extract on 4 MDR-M.tb isolates(No.13420,No.13449, No.13451 and No.13424), Mycobacterium intracellulare,Mycobacterium scrofulaceum, Mycobacterium fortuitum, Mycobacterium abscessus, Mycobacterium chelonae strains were 200 mg.mL-1, on MDR-M.tb and Mycobacterium bovis isolate No.13445 was 150 mg.mL-1,on Mycobacterium vaccae, Mycobacterium smegmatis, Mycobacterium avium, Mycobacterium gastri and Mycobacterium simiae strains were 100 mg.mL-1, and on Mycobacterium phlei, Mycobacterium gordonae and Mycobacterium kansasii strains were 50 mg.mL-1. Both water extract and alcohol extract of NBXH had different degree of inhibitory action on the Mycobacterium bovis, MDR-M.tb and NTM. The bacteriostatic action of NBXH ethanol extract was stronger than NBXH water extract.2. The bacteriostatic action of the traditional Chinese medicine compound NBXH extracts in vivoOne hundred and sixty 6-8 weeks old female BALB/c mice with weight 17-19 g were purchased, in which 8 mice used as normal control group, 152 mice were infected via tail vein with 5 × 104 CFUs of M.tb H37 Rv suspension to develop mouse tuberculosis(TB) model. The infected mice were weighed, randomly divided into the following 19 groups and then treated at the third day after infection: untreated control group, INH treatment group, M. vaccae treatment group, NBXH1 middle dose group, NBXH1 high dose group, NBXH2 middle dose group, NBXH2 high dose group, NBXH3 middle dose group, NBXH3 high dose group,NBXH4 middle dose group, NBXH4 high dose group, NBXH1 middle dose +INH group, NBXH1 high dose +INH group, NBXH2 middle dose+INH group, NBXH2 high dose + INH group, NBXH3 middle dose + INH group, NBXH3 high dose +INH group, NBXH4 middle dose + INH group,NBXH4 high dose + INH group. Taking the effective dose of adult clinical administration as the standard, the mouse’s dosages were determinedaccording to the weight conversion. Each mouse was given 0.5ml of NBXH and INH(containing 0.4 mg) by intragastric administration for 5times each week except Saturday and Sunday for 13 weeks. The mice in M.vaccae(22.5μg/100μl) treatment group were injected intramuscularly for6 times at 2 week intervals. The general condition and growth of body weight of the mice were observed weekly. The organ indexes of the lung,liver and spleen were calculated, and the lung tissue colony counting was performed each group at 13 weeks after treatment, to understand the therapeutic effect of NBXH against tuberculosis in mouse TB model.After treatment for 13 weeks, compared with the untreated control group, the body weight of mice significantly increased in the INH group,M. vaccae group, NBXH1 middle or high dose groups, NBXH2 middle or high dose groups, NBXH3 middle dose group, NBXH1 middle or high dose +INH groups, NBXH2 high dose +INH group, NBXH3 middle or high dose+INH groups and NBXH4 middle dose+INH group(P<0.05); the lung organ indexes significantly decreased in INH group, M. vaccae group,NBXH1 middle dose group, NBXH2 middle dose group, NBXH3 middle or high dose groups, NBXH4 middle dose group, NBXH1 middle or high dose+INH group, NBXH2 middle or high dose+INH group, NBXH3 high dose+INH group, and NBXH4 middle or high dose+INH group(P<0.05);the liver organ indexes significantly decreased in INH group, NBXH2 middle or high dose groups, NBXH3 high dose group, NBXH4 high dose group, NBXH2 middle or high dose+INH group and NBXH4 middle dose+INH group(P<0.05); the spleen organ indexes significantly decreased in INH group, NBXH3 middle or high dose group, NBXH4 middle dose group, NBXH1 middle or high dose +INH groups, NBXH2 high dose+INH group, NBXH3 middle or high dose+INH group and NBXH4 middle dose +INH group(P<0.05); the lung colony counting also significantly decreased in INH group, NBXH1 high dose group, NBXH2 middle dose group, NBXH4 high dose group and the of the combination of NBXH and INH groups(P<0.05). The traditional Chinese medicine compound NBXH extracts prepared by different processes in vivo could treat differently the mouse models of TB, improve the general condition in mouse, reduce the degree of pathological changes and the viscera colony count. The anti-tuberculosis effect of NBXH1 and NBXH2 were stronger than NBXH3 and NBXH4. The anti-tuberculosis effect of the part of the combination group were stronger than Chinese traditional medicine group alone, but the most of the combination group had no significant difference with the medicine group.