J147 modulates microglial polarization via CAMKK2/AMPK signaling to ameliorate neuroinflammation.

Neuroinflammation, characterized by an imbalance in microglial polarization, significantly contributes to various neurological conditions, including brain injuries, and represents a key therapeutic target for these conditions. This study aimed to determine whether J147, a known neuroprotective compound, could avert neuroinflammation by influencing microglial responses via CAMKK2/AMPK signaling. Employing an LPS-induced neuroinflammation model, we demonstrate that J147 exhibits strong anti-inflammatory effects in vivo, protecting brain structure and rebalancing cytokine profiles (e.g., suppressing TNF-α/IL-6 while enhancing IL-10). Cellularly, J147 induced a functional switch from M1 (iNOS+, CD40+) to M2 (CD206+, IL-10+) microglial phenotypes, consistent across both in vivo brain samples and in vitro microglial cultures, indicating a strong anti-inflammatory reprogramming effect. Additionally, J147 improved LPS-suppressed CAMKK2/AMPK phosphorylation. However, pharmacological inhibitors (STO-609/dorsomorphin) of the CAMKK2/AMPK signaling cascades abolished J147's neuroprotective effects, indicating that J147's neuroprotective potential is CAMKK2/AMPK-dependent. Complementary in vitro studies using microglia-oligodendrocyte co-cultures validated J147's dual functionality, restoring microglial viability (90 % recovery versus LPS) while promoting M2 polarization and maintaining oligodendrocyte integrity (85 % increase in MBP), all of which is contingent upon intact CAMKK2/AMPK signaling.