Enhanced autophagy and cholesterol efflux in mouse mesenchymal stem cells infected with H37Rv compared to H37Ra

Autophagy, metabolism, and associated signaling pathways play critical roles in bacterial survival within mammalian cells and influence the immunopathogenesis of infections. Mesenchymal stem cells (MSCs) are important host cells during Mycobacterium tuberculosis (Mtb) infection, yet how autophagy, metabolism, and related pathways are modulated in MSCs infected with the virulent H37Rv or the attenuated H37Ra strain of Mtb remains poorly understood. In this study, we utilized RNA-Seq screening, qRT-PCR, and Western Blotting to investigate the differences in these processes between H37Rv and H37Ra infections. Our results show that, at early time points (no more than 24h), infection with H37Rv significantly increased the expression of TlLR2, Prkaa2, and Prkaa2 phosphorylation in MSCs compared with H37Ra infection. Further analysis revealed that H37Rv infection induced a stronger autophagic response (evidenced by increased Atg9b and LC3II/LC3I) through the TLR2-AMP-AMPK pathway than H37Ra infection. Despite these differences in autophagy, there was no statistically significant difference in bacillary loads, suggesting that, in addition to autophagy, other factors such as apoptosis and immune-inflammatory responses may also regulate Mtb growth in MSCs. Additionally, the metabolic analysis showed that H37Rv infection led to increased expression of SLC2A3, PFKFB3, HK1, and ABCA1 in MSCs compared to H37Ra infection. These findings confirm that, during the early stages of infection, H37Rv induces enhanced autophagy, glucose metabolism, and cholesterol efflux through a more active TLR2-AMP-AMPK pathway than H37Ra. Therefore, MSCs may represent a novel target for the prevention and treatment of tuberculosis.