1-(2-4-dichlorobenzyl)indazole-3-carbohydrazide and Brain-Infarction

Neuroinflammation mediated by activation of microglia and interruption of the blood-brain barrier (BBB) is an important factor that contributes to neuron death and infarct area diffusion in ischemia reperfusion injury. Finding novel molecules to regulate neuroinflammation is of significant clinical value. We have previously shown that adjudin, a small molecule compound known to possess antispermatogenic function, attenuates microglia activation by suppression of the NF-κB pathway. In this study we continued to explore whether adjudin could be neuroprotective by using the transient middle cerebral artery occlusion (tMCAO) model. Adjudin treatment after reperfusion significantly decreased the infarction volume and neuroscore compared to the vehicle group. Staining of CD11b showed that adjudin markedly inhibited microglial activation in both the cortex and the striatum, accompanied by a reduction in the expression and release of cytokines TNF-α, IL-1β and IL-6. Concomitantly, adjudin noticeably prevented BBB disruption after ischemia and reperfusion, as indicated by the reduction of IgG detection in the brain cortex and striatum versus the vehicle group. This finding was also corroborated by immunofluorescence staining and immunoblotting of tight junction-related proteins ZO-1, JAM-A and Occludin, where the reduction of these proteins could be attenuated by adjudin treatment. Moreover, adjudin obviously inhibited the elevated MMP-9 activity after stroke. Together these data demonstrate that adjudin protects against cerebral ischemia reperfusion injury, and we present an effective neuroinflammation modulator with clinical potential.

Topics: Animals; Blood-Brain Barrier; Brain; Brain Infarction; CD11b Antigen; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Hydrazines; Indazoles; Matrix Metalloproteinase 9; Mice; Microglia; Neurologic Examination; Neuroprotective Agents; Nitric Oxide Synthase Type II; Occludin; Reperfusion Injury; Zonula Occludens-1 Protein

Neuroinflammation caused by microglial activation plays a key role in ischemia, neurodegeneration and many other CNS diseases. In this study, we found that Adjudin, a potential non-hormonal male contraceptive, exhibits additional function to reduce the production of proinflammatory mediators. Adjudin significantly inhibited LPS-induced IL-6 release and IL-6, IL-1β, TNF-α expression in BV2 microglial cells. Furthermore, Adjudin exhibited anti-inflammatory properties by suppression of NF-κB p65 nuclear translocation and DNA binding activity as well as ERK MAPK phosphorylation. To determine the in vivo effect of Adjudin, we used a permanent middle cerebral artery occlusion (pMCAO) mouse model and found that Adjudin could reduce ischemia-induced CD11b expression, a marker of microglial activation. Furthermore, Adjudin treatment attenuated brain edema and neurological deficits after ischemia but did not reduce infarct volume. Thus, our data suggest that Adjudin may be useful for mitigating neuroinflammation.

Topics: Analysis of Variance; Animals; Anti-Inflammatory Agents; Brain Edema; Brain Infarction; CD11b Antigen; Cell Line, Transformed; Cell Survival; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Interactions; Hydrazines; Indazoles; Infarction, Middle Cerebral Artery; Lipopolysaccharides; Male; Mice; Mice, Inbred ICR; Microglia; Nervous System Diseases; NF-kappa B; Psychomotor Performance; Signal Transduction; Time Factors