Network pharmacology and molecular docking-based analysis of protective mechanism of MLIF in ischemic stroke

Abstract

ObjectiveThis study aimed to evaluate the potential mechanism by which Monocyte locomotion inhibitory factor (MLIF) improves the outcome of ischemic stroke (IS) inflammatory injury.MethodsPotential MLIF-related targets were predicted using Swiss TargetPrediction and PharmMapper, while IS-related targets were found from GeneCards, PharmGKB, and Therapeutic Target Database (TTD). After obtaining the intersection from these two datasets, the Search Tool for Retrieval of Interacting Genes/Protein (STRING11.0) database was used to analyze the protein-protein interaction (PPI) network of the intersection and candidate genes for MLIF treatment of IS. The candidate genes were imported into the Metascape database for Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. The top 20 core genes and the “MLIF-target-pathway” network were mapped using the Cytoscape3.9.1. Using AutoDock Vina1.1.2, the molecular docking validation of the hub targets and MLIF was carried out. In the experimental part, transient middle cerebral artery occlusion (tMCAO) and oxygen and glucose deprivation (OGD) models were used to evaluate the protective efficacy of MLIF and the expression of inflammatory cytokines and the putative targets.ResultsMLIF was expected to have an effect on 370 targets. When these targets were intersected with 1,289 targets for ischemic stroke, 119 candidate therapeutic targets were found. The key enriched pathways were PI3K-Akt signaling pathway and MAPK signaling pathway, etc. The GO analysis yielded 1,677 GO entries (P < 0.01), such as hormone stimulation, inflammatory response, etc. The top 20 core genes included AKT1, EGFR, IGF1, MAPK1, MAPK10, MAPK14, etc. The result of molecular docking demonstrated that MLIF had the strong binding capability to JNK (MAPK10). The in vitro and in vivo studies also confirmed that MLIF protected against IS by lowering JNK (MAPK10) and AP-1 levels and decreasing pro-inflammatory cytokines (IL-1, IL-6).ConclusionMLIF may exert a cerebral protective effect by inhibiting the inflammatory response through suppressing the JNK/AP-1 signaling pathway.

Interplay between polygenic variants related immune response and lifestyle factors mitigate the chances of stroke in a genome-wide association study

“…RBFOX1 is a splicing factor of Ca/calmodulin-dependent protein kinase II γ ( CAMK2G ), and its expression is reduced in patients with post-myocardial infarction (28) . Monocyte locomotion inhibitory factor ( MLIF ) protects against ischaemic stroke by lowering MAPK10 (JNK3) and decreasing proinflammatory cytokines (29) . The DLG2 gene encoding postsynaptic density protei n -93 (PSD-93) and its inhibition activates nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) antioxidant signalling in oxygen-glucose deprivation and reoxygenation (OGD/R)-exposed neurons (30) .…”

Aquaporins: Gatekeepers of Fluid Dynamics in Traumatic Brain Injury

“…Through molecular docking techniques, AQP4 was pinpointed as one of the top ten central genes discovered in this study. Findings revealed that MLIF possesses anti-apoptotic properties and suppresses the expression of AQP4 protein, thereby playing a protective role in traumatic brain injury [ 155 ].…”

“…Antisense oligonucleotides Bind to mRINA, prevent AQP4 functional protein formation, reduce edema if administered early [145] Minocycline Suppresses AQP4 expression post-TBI, protects BBB integrity, modulates astrocyte characteristics [146] Acetazolamide Inhibits CA, prevents AQP4 redistribution post-TBI, mitigates cytotoxic edema [147] miR-211-5p Regulates MMP9/AQP4 axis, therapeutic potential for TBI treatment by targeting this pathway [148] Trifluoperazine Reduces AQP4 accumulation, mitigates brain edema, reduces neurological severity post-TBI [149][150][151] Adenine dinucleotide phosphate oxidase 2 NOX2 inhibition reduces AQP4 levels, enhances cognitive function, reduces brain edema post-TBI [152] Hypertonic saline Reduces AQP4, TNFa, IL-1B levels, and brain water contact, suppresses apoptosis post-TBI [153] Monocyte locomotion inhibitor factor Reduces brain water content, suppresses AQP4 suppression, provides protection against TBI [154,155] Lentivirus-mediated AQP4 gene silencing Reduces AQP4 expression, mitigates brain edema, reduces neurological deficits and neuronal apoptosis post-TBI [156] Omega-3 polyunsaturated fatty acids Improve glymphatic clearance, reduce AB42 accumulation, protect BBB integrity post-TBI [162] Angiotensin II type 1 receptor Deficiency preserves BBB integrity, reduces AQP4 polarization, improves glymphatic function, and AB clearance post-TBI [158] Melatonin receptors Interaction with estrogen may reduce BBB permeability, influence AQP4 expression [160] Arginin and laminin Regulate BBB integrity and AQP4 polarization, important for astrocyte migration and plasticity [161] Future research should focus on developing specific inhibitors and modulators of AQP channels, particularly AQP4, to directly intervene in the edema formation process. Additionally, the exploration of gene silencing techniques, such as the use of antisense oligonucleotides, and the modulation of aquaporin expression through pharmacological agents or dietary supplements like Omega-3 PUFAs could provide new therapeutic avenues.…”