NDRG4 overexpression is associated with reduced apoptosis after intracerebral hemorrhage via the PI3K/Akt/GSK3β signaling pathway

TL;DR

NDRG4 overexpression reduces brain cell death after intracerebral hemorrhage by activating a specific signaling pathway, improving neurological outcomes in rats.

Contribution

This study identifies NDRG4 as a novel modulator of apoptosis in ICH via the PI3K/Akt/GSK3β pathway.

Findings

NDRG4 overexpression improved neurological function and reduced brain water content after ICH.

NDRG4 reduced apoptosis markers like TUNEL-positive cells and cleaved caspase-3.

PI3K/Akt/GSK3β pathway activation was linked to NDRG4's protective effects.

Abstract

Intracerebral hemorrhage (ICH) is a severe form of stroke with high mortality, and apoptosis in the perihematomal region contributes to neurological deficits. This study aimed to investigate the role of NDRG4 in cerebral injury following ICH, focusing on apoptosis and related signaling pathways. A total of 242 male Sprague Dawley rats were used to establish a collagenase-induced ICH model and were allocated across four experiments to examine NDRG4 temporal expression, validate adenoviral overexpression, evaluate its effects on ICH outcomes, and probe PI3K/Akt/GSK3β signaling (6 rats per group). Neurological function, brain water content, TUNEL staining, Western blotting, and RT-qPCR were used to assess the effects of NDRG4 overexpression on ICH-induced brain injury and apoptosis. NDRG4 expression was significantly reduced in perihematomal brain tissue after intracerebral hemorrhage. In rats receiving adenoviral NDRG4 overexpression, neurological performance was significantly better than in ICH controls, and brain water content was significantly lower. NDRG4 overexpression was also associated with a significant reduction in TUNEL-positive cells, a significantly lower Bax/Bcl-2 ratio, and significantly decreased cleaved caspase-3 levels, while Bcl-2 levels were significantly higher. These biochemical and histological differences were accompanied by significantly increased phosphorylation of Akt (Ser473) and GSK3β (Ser9). Co-administration of wortmannin was associated with partial attenuation of these changes, suggesting that the observed effects may be related to activation of the PI3K/Akt/GSK3β signaling pathway. NDRG4 overexpression was associated with reduced perihematomal injury and improved neurological scores, partly associated with activation of the PI3K/Akt/GSK3β pathway. Further studies are warranted to delineate the specific cell types involved, the detailed mechanisms, and the translational relevance of these findings.

The online version contains supplementary material available at 10.1038/s41598-025-33247-5.

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