Transcriptome and metabolome profiling reveal the effect of MTB virulence protein Rv3874 and Rv3875 on macrophage Sphingolipid metabolism

Abstract Background Normal lipid metabolism is a key process to maintaining cellular immune function and energy metabolism of macrophages, but Mycobacterium tuberculosis (MTB) infection can cause lipid metabolism disorder of macrophages, resulting in granuloma or intracellular retention. Methods Recombinant BCG with virulence protein Rv3874 or Rv3875(rBCG-Rv3874 and rBCG-Rv3875)were constructed. Transcriptomics and High-performance liquid chromatography-mass spectrometry (HPLC-MS) metabonomics studies were performed using rBCG infected macrophage samples. Extensive and variation difference analyses were conducted on transcription and lipid metabolism levels compared with the control group. Results A fact is that 7100 genes were changed in the rBCG-Rv3874 group and 1263 genes were changed in the rBCG-Rv3875 group at the RNA-seq level. A total of 612 and 485 substances were detected in the non-targeted positive and negative ion mode of lipid metabolism, and a total of 1240 and 210 metabolites were detected in the targeted positive and negative ion mode of lipid metabolism. Finally, it was found that the lipid changes related to the sphingomyelin (SM) metabolic pathway were the most obvious. 220 kinds of SM metabolites were detected by positive ion detection, and it was found that more ceramide, ceramide-1-phosphate, and SM were enriched in the rBCG-Rv3874 group than in rBCG-Rv3875 group. Conclusion MTB virulence proteins Rv3874 and Rv3875 can affect the sphingolipid metabolism of macrophages and it's associated with phagocytosis, apoptosis, and inflammatory processes.