Immunoproteasome Inhibition Modulates Microglial Polarization to Facilitate Anti-Inflammatory Responses and Hematoma Resolution After Intracerebral Hemorrhage.

Abstract

Intracerebral hemorrhage induces severe secondary brain injury characterized by excessive neuroinflammation and inefficient hematoma clearance, processes largely governed by microglial polarization and phagocytic activity. The immunoproteasome, an inducible proteasome isoform involved in immune regulation, has been implicated in inflammatory neurological disorders, but its role in microglial responses after ICH remains unclear. In this study, rat models of common hemorrhage, severe hemorrhage, and severe hemorrhage with hematoma aspiration were used to represent graded injury severity and post-evacuation recovery. Transcriptomic profiling at day 3 post-injury identified immunoproteasome-associated gene networks, while expression of the catalytic subunits LMP2 and LMP7, microglial polarization markers, and phagocytic receptors was analyzed by Western blotting and immunofluorescence. Severe hemorrhage markedly induced LMP2 and LMP7 expression, predominantly in Iba1microglia, accompanied by enhanced ER stress, NF-κB signaling, and M1-like polarization and reduced phagocytic marker expression. Hematoma aspiration attenuated immunoproteasome expression and restored M2-associated and phagocytic signatures. Consistently, pharmacological inhibition of immunoproteasomes in primary microglia enhanced erythrophagocytosis and promoted a reparative phenotype in vitro. These findings indicate that immunoproteasome activation links hemorrhagic severity to maladaptive microglial polarization and impaired hematoma clearance after ICH, and that reducing immunoproteasome expression may help rebalance inflammatory and phagocytic microglial functions.