Linoleic Acid-Lyso PG Axis promoting lipid droplet–mitochondria tethering by stabilizing Noncanonically Mitochondrial PPAR β/δ to Ameliorate Microglial Dysfunction in subarachnoid hemorrhage

Background

Microglial lipid handling and mitochondrial failure contribute to brain injury after subarachnoid hemorrhage (SAH), but the lipid signals coupling these processes remain unclear. We investigated whether linoleic acid (LA) restores microglial homeostasis through lysophosphatidylglycerol 16:0 (LPG[16:0]) and peroxisome proliferator-activated receptor-δ (PPARδ).

Methods

Case-control CSF metabolomics included 30 patients with aneurysmal SAH and 10 controls. Mechanisms were examined in a blood-injection mouse model and hemoglobin-exposed primary mouse microglia using targeted lipidomics, RNA sequencing, mitochondrial and phagocytosis assays, pharmacological perturbation, fractionation, coimmunoprecipitation, thermal shift analysis, and structural modeling. Behavioral outcomes were evaluated by open- field, Y-maze, and Morris water-maze testing.

Results

CSF LA concentrations were higher in patients with SAH than in controls and discriminated the groups within this cohort (area under the curve, 0.9967 [95% CI, 0.9859–1.000]; P<0.001). LA attenuated inflammatory activation and restored phagocytosis, mitochondrial membrane potential, respiration, and ATP production in hemoglobin-exposed microglia. LA restored PLA2G15-associated LPG(16:0) levels, which phenocopied these effects.

Transcriptomic and inhibitor analyses identified PPARδ as a downstream effector. LPG(16:0) increased PPARδ stability, and fractionation and protease protection identified a PPARδ pool on the cytosolic face of the outer mitochondrial membrane. PPARδ associated with PLIN2 and CPT1A, promoted lipid droplet–mitochondria apposition, and supported fatty acid oxidation. In mice, LA reduced neuroinflammatory injury and partially improved anxiety- related behavior and spatial memory.

Conclusions

The LA–LPG(16:0)–PPARδ axis links glycerophospholipid remodeling to organelle coupling and mitochondrial recovery in microglia after SAH. The preventive dosing paradigm requires validation in clinically relevant treatment settings.