Neuroprotection of probucol following intracerebral hemorrhage in mice through anti-inflammation and blood-brain barrier protection

Background Intracerebral hemorrhage (ICH) is a disease with high mortality and morbidity worldwide, yet pharmacotherapies for ICH are currently lacking. Probucol has gained widespread attention in recent years for its anti-free radical, anti-inflammatory, and lipid metabolism regulatory properties. Its potential value in neuroprotection has been extensively studied. Yet, the mechanisms and role of its neuroprotection in intracerebral hemorrhage (ICH) are still not well understood. The present investigation seeks to examine probucol’s neuroprotective impacts the ICH model. Methods In this study, ICH was induced in mice via type VII collagenase injection into the basal ganglia, and mice received daily probucol treatment (60 mg/kg/day) for three consecutive days after ICH modeling. At 3 days post-ICH modeling, a panel of experimental assays, including measuring the volume of hematoma, behavioral function tests, Western blot analysis, immunofluorescence staining, dry and wet weight, Evans blue test and terminal deoxynucleotidyl transferase-dUTP nick end labeling (TUNEL) staining were conducted to assess neurological deficits, microglial and neutrophil-associated neuroinflammation, cerebral edema, tight junction protein expression and blood-brain barrier (BBB) leakage. Results Detailed analysis of hematoma tissues demonstrated that probucol diminishes hematoma volume and improves neurological function in post-ICH mice. Immunofluorescence assays showed that probucol suppresses neutrophil infiltration, microglia/macrophage activation, and pro-inflammatory cytokine secretion. Moreover, probucol markedly downregulates matrix metalloproteinase-9(MMP-9) expression while upregulating the levels of tight junction proteins zonula occludens protein-1 (ZO-1) and Occludin. Probucol also reduced blood-brain barrier (BBB) permeability, as evidenced by decreased brain water content and attenuated Evans blue extravasation, thereby ameliorating BBB structural damage. In addition, probucol upregulated the expression of anti-apoptotic B-cell lymphoma 2 (Bcl-2) and inhibited the expression of pro-apoptotic Bcl-2 associated X protein (Bax), ultimately reducing the number of TUNEL-positive dead neuronal cells in ICH brain tissue. Conclusion This study confirms that probucol can suppress excessive neuroinflammation, maintain the structural integrity of the blood-brain barrier and inhibit neuronal apoptosis after ICH, which provides novel experimental evidence and a potential research direction for the development of targeted clinical drugs for ICH treatment.