Mechanism and influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage

Objective  To study the influence of down-regulation of miR-29a-3p expression on apoptosis in mouse macrophage and expression of apoptosis-associated genes, and provide references for investigating the pathogenesis mechanism of mycobacterium tuberculosis (MTB) and developing new diagnosis and treatment methods.

Method  The recombinant inhibitor vector pEZX-AM02-miR-29a-3p was transfected into RAW264.7 cells. Transfection efficiency was observed through fluorescence microscopy, the expression levels of miR-29a-3p and apoptosis-associated genes caspase3, caspase7, caspase8, Bcl-2, Mcl-1 and Bax were detected by RT-PCR, and the apoptosis rates of RAW264.7 cells were detected by flow cytometry at 24, 36, 48 h after transfection.

Result  After transfected with pEZX-AM02-miR-29a-3p recombinant inhibitor vector, miR-29a-3p expression in RAW264.7 cells decreased significantly, while the target genes caspase7, caspase8, Bcl-2, Mcl-1 and Bax were up-regulated at different levels. The results of flow cytometry indicated that the apoptosis rates increased as the transfection time progressed and reached the peak at 36 h.

Conclusion  The expression of miR-29a-3p in mouse macrophage are inhibited by transfecting with pEZX-AM02-miR-29a-3p, which promotes the apoptosis of macrophage by up-regulating the expression of target genes, suah as caspase7, caspase8, Bcl-2 and Mcl-1.