angiogenin and Infarction--Middle-Cerebral-Artery

Stroke is a devastating event with a limited choice of intervention. Benzoinum is frequently used to treat stroke in traditional Chinese medicine. Our team has found that the neuroprotection of benzoinum may related to angiogenesis, but the exact biological mechanism is unclear. The objective of this study was to explore its biological mechanism of angiogenesis in cerebral ischemia model rats.. First, network pharmacology and molecular docking were performed to predict the possible targets and mechanisms of benzoinum in treating ischemic stroke. The best dose was then selected according to pharmacodynamic indexes such as those for neurological deficit, cerebral infarction rate, and brain histopathology in middle cerebral artery occlusion (MCAO) model rats. Finally, RT-PCR, Western Blot and immunohistochemical analysis were applied to verify the prediction results from molecular docking.. Network pharmacology and molecular docking demonstrated that the targets of treating cerebral ischemia were PDE4D, ACE and TTR, and the mechanism may be related to the ACE-AngI-VEGF signaling pathway. Experimental verification results suggested that 0.50 g/kg and 1.00 g/kg benzoinum could significantly protect against neurological deficit and reduce cerebral infarction rate in the cerebral cortex and hippocampus in MCAO model rats. At an optimal dose, benzoinum could significantly up-regulate VEGF, SHH and ANG-1, yet down-regulate ACE expression in MCAO model rats.. Balsamic acid is the active ingredient of benzoinum that protects against ischemic stroke and the possible mechanism is related to the promotion of angiogenesis via regulating ACE-AngI-VEGF pathway.

Topics: Animals; Brain Ischemia; Cerebral Cortex; Disease Models, Animal; Drugs, Chinese Herbal; Gene Expression Regulation; Infarction, Middle Cerebral Artery; Male; Molecular Docking Simulation; Neovascularization, Pathologic; Neuroprotective Agents; Peptidyl-Dipeptidase A; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Ribonuclease, Pancreatic; Small Molecule Libraries; Stroke; Vascular Endothelial Growth Factor A

Angiogenin (ANG) is a potent angiogenic factor. The purposes of this study were to observe the change of the expression level of ANG and to identify the cell types that express ANG in the focal ischemic rat brain. The rat brain ischemia-reperfusion model was produced by a 2 h occlusion of the left middle cerebral artery with a nylon thread followed by reperfusion for 1 day, 3 days, 7 days or 14 days. The expression levels of ANG in the rat brain at each time points were determined by western blotting. The co-staining of ANG with NeuN was observed using double immunofluorescent labeling combined with confocal laser scanning microscope. We found that the expression level of ANG increased significantly in the rat brain 1 day, 3 days, and 7 days after ischemia (P<0.05), and peaked 3 days after ischemia. Double immunofluorescent labeling showed that ANG positive cells were mostly co-stained with NeuN. Our observations suggest that the level of ANG increased significantly in the rat brain after ischemia. The upregulated ANG was mostly expressed by neurons.

Topics: Analysis of Variance; Animals; Antigens, Nuclear; Blotting, Western; Brain; Brain Ischemia; Fluorescent Antibody Technique; Infarction, Middle Cerebral Artery; Male; Microscopy, Confocal; Nerve Tissue Proteins; Neurons; Random Allocation; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Ribonuclease, Pancreatic; Time Factors