Transcriptome analysis reveals the effect of STAT3 gene of JAK-STAT signaling pathway on apoptosis of Brucella macrophages

Brucella is a significant intracellular parasitic bacterium that establishes an infection within a host cell to facilitate its proliferation. While the survival of Brucella in cells is an important manifestation of its virulence, the research on this aspect is not yet complete. Transcriptome sequencing techniques were employed for the analysis of transcript levels of various genes in RAW 264.7 macrophages infected with M5 and wild strains. siRNA can be employed to silence differentially expressed genes, with the subsequent validation of their functions conducted through a variety of techniques, including RT-qPCR, Western blot, flow cytometry, and others.In the study, a significant up-regulation of the STAT3 gene was observed in the vaccine strain when compared with the wild strain. The design of the relevant siRNA to silence STAT3 resulted in the promotion of apoptotic gene expression and the inhibition of apoptosis-inhibiting gene expression, thereby facilitating cell apoptosis.The results demonstrated that the discrepancy in STAT3 gene transcription between the vaccine and wild strains resulted in a reduction in cell damage and a decline in the virulence of the vaccine strain M5. This finding offers novel insights into the intracellular survival mechanism of Brucella and the underlying cause of M5 weakness. Overall design: RNA seq analysis was performed on RAW 264.7 macrophages infected with M5 and wild-type strains.