Study On The Relationship Between The Molecular Action Of Feilaokang Anti-isoniazid Resistant Tuberculosis And KatG Gene Mutation

Objective:Through the adoption of whole genome sequencing methods to st-udy whether the molecular action that FeiLaoKang resists isoniazid resista-nce Mycobacterium tuberculosis associating with KatG gene Mutation is related to r everse mutation or gene repair,provides new train of thought for the trea-tmen t of TCM of Isoniazid-resistant tuberculosis.Methods:1.Grouping of experimental animals:120 Kunming mice of SPF grade, initial weight 18-22g, were randomly divided into 6 groups and there were 20 mice in each group, respectively for standard bacteria group..resistance INH group, FLK group, INH group and the combined group.2.Model.-Blank group was not injected into tail vein;standard bacteria group the mice via tail vein were given by injection of H37Rv standard strains of Mycobacterium tuberculosis?ATCC:27294?; The mice of the other four groups viatail vein were given by injection resistance to isoniazid katG gene mutations in Mycobacterium tuberculosis strains of bacteria suspension.3. Drug intervention:The mice of standard bacteria group and resistant to isoniazid group were given by gavage with physiological saline for 56 days, once a day. The mice of the other three groups respectively were given by gavage with FLK liquid, isoniazid solution and feilaokang combined with isoniazid mixture for 56 days, once a day.4. After the end of the experimental period, the mice were isolated from the lung tissue for bacterial culture and pathological examination. The lung tissue of 1-2 mice was randomly selected from each group for the purification and sequencing of Mycobacterium tuberculosis DNA.Results:1.The bacterial cultures of the mice were positive in others groups except blank group.2. Through the observation of the pathological morphology in lungs of mice, we found that pharmacotherapy to mouse lung tissue pathological changes compared with the wild MTB infection mice and isoniazid resistant MTB infection g-roup light, such as hyperplasia lung tissue, exudation, necrosis;3.The position of 2155168 and 2154724is the R463L and S315T of KatG gene in the Chromosome?NC_<sub>000962.3?,where in addition to the wild strains group, the rest of the four groups are gene mutation; using the method of sequencing to detecte the isoniazid resistance MTB chromosomes, We found the Rv0104, Rv2079,and Rv3898c as termination codon mutations, which are encoded hypothetical protein gene, as well as the Rv3327 gene mutations that is the gene of coding transposase fusion protein; After treatment with FLK and FLK joint INH, we found the RV0063 gene mutations into termination codon in chromosome?NC₀00962? which coding transposase fusion protein through isoniazid resistant MTB gene sequencing.Conclusion:1.Successively establish the mycobacterium tuberculosis infect-ed mice model; 2.In this study we found that gene sequencing of KatG protein showed no reverse mutation in the 2 amino acids different from those of wil-d strain, after FLK therapy; 3.The gene sequencing of Rv0104, Rv2079, Rv38-98c and Rv3327 protein showed mutation after drug therapy which code hypot hetical protein and transposase fusion protein, this result explains the drug the rapy that probably could affect INH resistance MTB gene expression; 4.The eff ect of FLK on isoniazid resistance MTB may be related to the mutation of R-V0063 gene to the termination codon.which can code Possible oxidoreductase.