Study On The Regulation And Mechanism That Mycobacterium Tuberculosis Reacts At The Apoptotic Of Host Macrophage

Objecve:To establish the models of mycobacterium tuberculosis infecting RAW264.7macrophages, and explore the regulation and mechanism of apoptosis of the macrophage infectedwith mycobacterium tuberculosis.Methods:1. Infected the macrophage RAW264.7cell line with international standards withMycobacterium tuberculosis H37Rv and BCG, at the same time, set the blank controlgroup, after the infection at the1st h,6th h,12th h, and24th h, flow cytometry wereemployed to detect the rate of the apoptosis of macrophages of each group.2. Then detect the caspase-3and Bcl-2protein and expression levels of the gene withwestern blot and RT-PCR.Results:1. The rate of the apoptosis of macrophage RAW264.7cell line infected by Mycobacteriumtuberculosis is significantly higher than that of the control group, the difference wasstatistically significant (P <0.05), and the rate of apoptosis of macrophage RAW264.7cell line infected by the BCG was higher than that of the H37Rv, after infection, at the1st h,12th h and24th h the rate of the apoptosis increased significantly, and thedifference was statistically significant (P <0.05).2. After being infected by Mycobacterium tuberculosis, the expression of the Caspase-3protein in macrophages increased, and the expression of the Caspase-3protein of theMycobacterium tuberculosis group is higher than the control group, the control group<H37Rv infection group <BCG infection group. The difference was statisticallysignificant (P <0.05). The expression of Caspase3protein in BCG infection group ishigher than that in H37Rv infection group, at the1st h,12th h and24th h, the expressionof protein was significantly increased after being infected, the difference was statisticallysignificant (P <0.05).3. The expression level of Caspase-3gene in the Mycobacterium tuberculosis infectiongroup was relatively higher than that in the control group, the difference was statisticallysignificant (P <0.05). The expression level of Caspase-3gene in the BCG infectiongroup was higher than that in the H37Rv infection group, at the1st h and24th h, theexpression level of Caspase-3gene increased significantly after infection, and thedifference was statistically significant (P <0.05).4. The expression level of Bcl-2gene in the Mycobacterium tuberculosis infection groupwas relatively higher than that in the control group. The difference was statisticallysignificant (P <0.05). After being infected, at the1st h,6th h and24th h. The expressionof Bcl-2protein in the H37Rv infection group was higher than that in the BCG infectiongroup, the difference was statistically significant (P <0.05).5. The expression level of Bcl-2gene in the Mycobacterium tuberculosis infection groupwas relatively higher than that in the control group. The difference was statisticallysignificant (P <0.05). The expression of Bcl-2protein in the H37Rv infection group washigher than that in the BCG infection group, the difference was statistically significant(P <0.05). After being infected, at the1st h,6th h and24th h. the expression of Bcl-2protein was significantly increased Conclusions:1. The rate of the apoptosis of macrophage was significantly increased after infected withMycobacterium tuberculosis.the intensity of virulence of Mycobacterium tuberculosiswas related to the apoptosis of cells.there was differience in the level of the rate of theapoptosis of macrophage. The apoptosis of macrophage infected with Mycobacteriumtuberculosis of weak poison was more significantly.2. There was differience in the level of the expression of the Caspase-3protein inmacrophages. The expression of the Caspase-3protein in macrophages was moresignificantly after infected with Mycobacterium tuberculosis of weak poison.3. There was differience in the level of the expression of the Bcl-2gene in macrophages.The expression of the Bcl-2gene in macrophages was more significantly after infectedwith Mycobacterium tuberculosis of weak poison.4. The intensity of virulence of Mycobacterium tuberculosis was related to the apoptosis ofmacrophages. And the expression of the Caspase-3protein and the Bcl-2gene inmacrophages.