| Objective: Tuberculosis(TB) is a chronic infectious diseases which induced by Mycobacterium tuberculosis(Mtb). During the tuberculosis infection, Mtb would be phagolysised by macrophages, then surviveing and proliferating inside. At last, Mtb result in the apoptosis of macrophages and causes TB. So what needed in prevention and treatment of TB is the deeper understand of mechanism of TB. To study the pathogenesis of TB and relationship between different virulence mycobacteria and infection, the genes related to inflammation and microRNAs related to the regulation of these genes were selected. Then the expression of the genes and microRNAs in macrophages which were infected by different virulence mycobacteria were detected. The data laid a foundation for the further exploration on the relationship between Mtb and macrophages.Method: This study select the induced THP-1 cell by different virulence of mycobacterium tuberculosis infected after couse the change of immune-relevant genes IL-1、IL6R、RAB6B and TLR4. Tatal RNA was extracted at different times(12h,24 h,36h). Detected these four target genes use the method of real-time PCR. According to the prediction of miRNA target genes selected via mi RNA relational database, which is microRNA-21、microRNA-15 and microRNA-29 a. Small RNA reverse transcribed cDNA by method of tail. The expression of microRNA-21、microRNA-15 and microRNA-29 a were detected by real-time PCR.Result: 1.The expression of four genes know to be related to innate immunity in THP-1 macrophage cells following H37 Rv, H37 Ra and BCG infection determined by real-time PCR. The expression of target IL-1 and TLR4 were up-regulation in every experimental group vs control,except that an decrease expression of target RAB6 B. However, the IL6 R was found down-regulated at 12 h with H37 Rv infection(P<0.05), an increaced expression of IL6 R was found after 24 h the infection. The expression of IL6 R was significantly increased at 12 h in BCG infection(P<0.05).2.The expression profile of three miRNAs known to be correlation with innate immune system.The expression of microRNA-21, microRNA-155 and microRNA-29 a were significantly increased, except microRNA-21 down-regulation after 12 h in BCG effection group. The expression of miRNA-21 was significantly increased at 36 h in BCG infection(P<0.05); The expression of miRNA-155 and miRNA-29 a were significantly increased(P<0.05).Conclusion: Three different kinds of mycobacterium tuberculosis were H37 Rv, H37 Ra and BCG, which infected macrophage respectively(12h, 24 h, 36h), that infected macrophage arouse the expression of immuno-related genes and microRNAs take up-regulation, except that an decrease expression of them. |
PDF Full Text Request