Correlation Study between Hsp16.3 of Mycobacterium tuberculosis and the apoptosis of infected macrophages

Objective To establish the models of Mycobacterium tuberculosis(MTB) infecting RAW264.7 macrophages,and study the correlation between Hsp16.3 of MTB and the apoptosis of infected macrophages.Methods RAW264.7 cells were infected by H37Rv and BCG strains,after being infected of 1 h,3 h,6 h,and 12 h,the infected RAW264.7 macrophages models were identified by confocal laster scanning microscope.Then flow cytometry was used to detect the macrophage apoptosis rate and analysis infected macrophages group apoptosis rate's change in different time phases.After PCR amplifying model purpose gene,western blot was used to compare and analysis the Hsp16.3 levels of phase changes.Results The infected RAW264.7 macrophages were identified by Confocal laster scanning microscope: MTB16KD green mark was detected.The mycobacterium tuberculosis infecting macrophages models were established.After being infected by H37Rv and BCG,the apoptosis rate of macrophages increased.The apoptosis rate in 1 、3 and 12 h increased obviously,and the difference was statistically significant(P0.05).With macrophages being infected by MTB,there was a specific band at 435 bp of PCR amplification products.The expression of MTBHsp16.3 increased after infection,which increased obviously in 1 h and 3 h.Conclusion The infection of MTB induced the macrophages' apoptosis;the apoptosis rate was related to the virulence of MTB;macrophages which were infected by the weak virulent MTB had a much more pronouned early apoptosis.Being infected by different virulence of MTB,the apoptosis rate of macrophages was related to the expression levels of MTB Hsp16.3 of which the expression of the power full virulent MTB was much more significant than that of the less virulent in the early phase of infection.