Effect Of MiRNA-29B And Caspase-3 And Caspase-7 In A Mouse Model Of Feilaokang Treat Isoniazid Resistant Tuberculosis

Objective:Detect the expression of caspase-3, caspase-7 and miRNA-29b in a mouse model of feilaokang treat Isoniazid resistant tuberculosis. Investigate the mechanism of feilaokang treat isoniazid resistant tuberculosis, and provide new ideas for the mechanism of traditional Chinese medicine in the treatment of drug resistant tuberculosis.Methods:1.Establish the isoniazid resistant tuberculosis SPF kungming mouse model of using intravenous injection of prepared isoniazid resistant Mycobacterium tuberculosis bacterium suspension,then use physiological saline,feilaokang,INH, feilaokang combined with INH 56 days by intragastric administration. Establish the TB mouse model of using intravenous injection of MTB bacterium suspension,then use physiological saline 56 days by intragastric administration. Use physiological saline 56 days by intragastric administration for mouse in the blank group.2.Detect expression level of caspase-7 and Caspase-3 in lung tissue with immunohistochemistry method.3.Detect expression level of miRNA-29b in lung tissue by real time quantity-PCR technology.4.The experimental results were analyzed statistically by SPSS 17.0 statistics software.Result:1.blank group did not detected MTB in mouse lung tissue. Both isoniazid resistant tuberculosis model group and the standard TB model group were detected MTB in mouse lung tissue.2.According to the immunohistochemical evaluation standard,compared with the blank group in mouse lung tissue,caspase-3 express high in standard TB model group(i'<0.01) and isoniazid resistant tuberculosis model group(P<0.05). After treatment of feilaokang,the expression of caspase-3 was higher than that of in the blank group(.P<0.05)but decreased than before treatment. After treatment of feilaokang combined with isoniazid,the expression of caspase-3 had no significant difference compared with the blank group(P> 0.05).After treatment of isoniazid,the expression of caspase-3 was higher than that of in blank group(P< 0.05).3.According to the immunohistochemical evaluation standard,compared with the blank control group in mouse lung tissue,caspase-7 express high in standard TB model group(P<0.01) and isoniazid resistant tuberculosis model group(P<0.01). After treatment of feilaokang combined with isoniazid, the expression of caspase-7 had no significant difference compared with the blank group(P>0.05).After treatment of feilaokang,the expression of caspase-7 had no significant difference compared with the blank group(P>0.05). After treatment of isoniazid,the expression of caspase-7 is still high ang had no significant difference compared with the blank group(P>0.05).4.The expression level of MiRNA-29b was lowest in blank group of mouse lung tissue.Compared with the blank group,the expression of MiRNA-29b increased in the standard TB group(P<0.01) and isoniazid resistant tuberculosis model group(P< 0.01),while in the isoniazid resistant tuberculosis model group were the highest.After treatment of feilaokang, the expression of miRNA-29b is lower than that in isoniazid resistant tuberculosis model group(P<0.01); After treatment of INH, the expression of miRNA-29b is lower than that in isoniazid resistant tuberculosis model group(P< 0.01); After treatment of feilaokang combined with INH, the expression of miRNA-29b is lower than that in isoniazid resistant tuberculosis model group(P< 0.01).Among them,after treatment of feilaokang,feilaokang combined with INH,the expression of miRNA-29b decreased than that after treatment of INH.After treatment of feilaokang, feilaokang combined with INH, the expression of miRNA-29b are similar to each other, and are lower than that in the standard TB model group.Conclusion:1.Inject isoniazid resistant Mycobacterium tuberculosis bacteria suspension by the mice tail vein.7 days later, MTB were detected in lung tissue of mouse.That is to say,the isoniazid resistant tuberculosis mouse model establish successfully.2.With the immunohistochemical method,the caspase-3 expression increased and decreased after treatment of feilaokan in the lung tissue of isoniazid resistant tuberculosis group.Mechanism of Feilaokang treat isoniazid resistant tuberculosis may probably via cell apoptosis which induced by caspase-3.3.With the immunohistochemical method,the caspase-7 expression increased and decreased after treatment of feilaokan in the lung tissue of isoniazid resistant tuberculosis model group.Mechanism of Feilaokang treat isoniazid resistant tuberculosis may probably via cell apoptosis which induced by caspase-7.4.The MiRNA-29b expression was increased in the lung tissues of isoniazid resistant tuberculosis model group,while decreased after treatment of feilaokang.Mechanism of Feilaokang treat isoniazid resistant tuberculosis may probably via the decrease of miRNA-29b.