12/15-lipoxygenase inhibition attenuates neuroinflammation by suppressing inflammasomes

IntroductionLipoxygenases (LOXs) have essential roles in stroke, atherosclerosis, diabetes, and hypertension. 12/15-LOX inhibition was shown to reduce infarct size and brain edema in the acute phase of experimental stroke. However, the significance of 12/15-LOX on neuroinflammation, which has an essential role in the pathophysiology of stroke, has not been clarified yet.MethodsIn this study, ischemia/recanalization (I/R) was performed by occluding the proximal middle cerebral artery (pMCAo) in mice. Either the 12/15-LOX inhibitor (ML351, 50 mg/kg) or its solvent (DMSO) was injected i.p. at recanalization after 1 h of occlusion. Mice were sacrificed at 6, 24, and 72-h after ischemia induction. Infarct volumes were calculated on Nissl-stained sections. Neurological deficit scoring was used for functional analysis. Lipid peroxidation was determined by the MDA assay, and the inflammatory cytokines IL-6, TNF-alpha, IL-1beta, IL-10, and TGF-beta were quantified by ELISA. The inflammasome proteins NLRP1 and NLRP3, 12/15-LOX, and caspase-1 were detected with immunofluorescence staining.ResultsInfarct volumes, neurological deficit scores, and lipid peroxidation were significantly attenuated in ML351-treated groups at 6, 24, and 72-h. ELISA results revealed that the pro-inflammatory cytokines IL-1beta, IL-6, and TNF-alpha were significantly decreased at 6-h and/or 24-h of I/R, while the anti-inflammatory cytokines IL-10 and TNF-alpha were increased at 24-h or 72-h of ML351 treatment. NLRP1 and NLRP3 immunosignaling were enhanced at three time points after I/R, which were significantly diminished by the ML351 application. Interestingly, NLRP3 immunoreactivity was more pronounced than NLRP1. Hence, we proceeded to study the co-localization of NLRP3 immunoreactivity with 12/15-LOX and caspase-1, which indicated that NLRP3 was co-localized with 12/15-LOX and caspase-1 signaling. Additionally, NLRP3 was found in neurons at all time points but in non-neuronal cells 72 h after I/R.DiscussionThese results suggest that 12/15-LOX inhibition suppresses ischemia-induced inflammation in the acute and subacute phases of stroke via suppressing inflammasome activation. Understanding the mechanisms underlying lipid peroxidation and its associated pathways, like inflammasome activation, may have broader implications for the treatment of stroke and other neurological diseases characterized by neuroinflammation

Benzoylcholine, Indole-3-acetic And Human Serum Butyrylcholinesterase Interactions

In this study we studied the kinetic interaction of indole-3- acetic acid with human serum butyrylcholinesterase. Butyrylcholinesterase was purified from human serum and the benzoylcholine kinetics was analyzed. The enzyme had a Km value of 14.22 ± 2.97 μM. The Hill plot was linear with a value of nH = 0.95, displaying that the enzyme follows Michaelis-Menten kinetics with regard to benzoylcholine. Through inhibition studies, indole-3-acetic acid was found to be a noncompetitive inhibitor for human serum butyrylcholinesterase with benzoylcholineas substrate. The Ki value was calculated as 1.86 ± 0.27 mM. In our previous study on human serum butyrylcholinesterase using butyrylthiocholine as substrate, indole-3-acetic acid was found to be a linear-mixed type inhibitor.This difference in kinetic behavior with regards to two different substrates could arise from the tighter binding of the more bulky and hydrophobic benzoyl group to the hydrophobic pocket of the active site gorge rather than the butyryl group and that the binding of this butyryl group could be sterically hindered by the indole-3-acetic acid

Counter-regulation of cholinesterases: differential activation of PKC and ERK signaling in retinal cells through BChE knockdown.

The ubiquitous cholinesterase (ChE) enzymes, functioning in the termination of acetylcholine mediated neural transmission, are also reported to have additional functions. Through application of siRNAs against butyrylcholinesterase (BChE) in R28 cells, a retinal cell line with pluripotent properties, a counter-regulation between ChEs was revealed. BChE knock down resulted in an up-regulation of not only acetylcholinesterase (AChE), but also altered the signaling status of PKC and ERK. Knockdown of BChE modified ERK signaling most notably through ERK1/2 proteins, together with the transcription activator P90RSK1 and c-fos. Stimulation of the R28 cell line by forskolin revealed that ChEs are involved in an intricate cross talk between different signaling pathways. Forskolin-stimulated R28 cells displayed a robust cholinergic response, as detected by both electrophysiology and ChE expression, and changed the activation status of PKC/ERK signaling pathways. The findings in R28 cells show that ChE expressions are inversely co-regulated and act through the transcription factors c-fos and P90RSK1. Since R28 cells have the capacity to differentiate into different cell types through stimulation of signaling pathways, ChEs are likely to be associated with cell fate determination, rather than just terminating cholinergic responses

Effects of conjugated linoleic acid supplementation and exercise on post-heparin lipoprotein lipase, butyrylcholinesterase, blood lipid profile and glucose metabolism in young men

This study was designed to investigate the effects of conjugated linoleic acid (CLA) supplementation and endurance exercise training-induced changes on post-heparin lipoprotein lipase (PH-LPL) and butyrylcholinesterase (BChE) activities along with leptin, insulin and lipid levels in plasma by a randomized double blind experiment. Eighteen sedentary male volunteers were randomly divided into CLA and Placebo (PLC) supplementation groups. Both groups underwent daily supplementation of either 3 g CLA or 3 g placebo for 30 days, respectively, and performed exercise on a bicycle ergometer 3 times per week for 30–40 min at 50% VO2 peak workload. For plasma glucose, insulin and leptin levels and BChE activity fasting blood was used. For PH-LPL measurements, blood was collected 15 min after 50 IU/kg iv heparin injection. In all groups, there is a statistically significant decrease in BChE (p = 0.03, p = 0.02) and leptin (p = 0.002), insulin and HOMA-IR levels (p = 0.02). Exercise with or without CLA supplementation decreased insulin levels and increased insulin sensitivity. PH-LPL activity was increased significantly in both groups, displaying increased fatty acid mobilization. We conclude that though CLA supplementation and exercise can affect these parameters, CLA is not more effective than exercise alone. Hence, a prolonged supplementation regime may be more effective. Taken together in our small study group, our findings display that BChE is a potential marker for synthetic function of liver, fat metabolism, an obesity marker, a function long overlooked

Effect of Asiatic Acid on The Treatment of Spinal Cord Injury: an Experimental Study in Rats

AIM: Spinal cord injury (SCI) is a devastating condition of the central nervous system. There is no proven therapeutic agent for the treatment of this complex disorder. Asiatic acid (AA) has been used as an anti-inflammatory and anti-oxidant agent in Eastern countries for many years. The aim of this study was to investigate the effectiveness of AA on the treatment of traumatic SCI in rats. MATERIAL and METHODS: Thirty-two adult male Sprague-Dawley rats were divided into 4 groups as laminectomy, laminectomy+trauma, vehicle, and M treatment groups. SCI was created by the modified Allen's weight-drop technique. After the injury, the levels of pro-inflammatory cytokines (IL-6, IL-beta, TNF-alpha) and lipid peroxidation products (MDA) were measured. Tarlov functional recovery scores were also determined for each rat. The One-way ANOVA test was used for the analysis of difference between 4 experimental groups and the groups were compared individually by Tukey-LSD post hoc analysis test (p=0.001). RESULTS: AA administration just after SCI attenuated the levels of lipid peroxidation products (MDA) and pro-inflammatory cytokines (TNF-alpha,IL1 beta). It also increased the Tarlov functional recovery scores of the rats. CONCLUSION: AA administration could attenuate a number of deleterious reactions after traumatic SCI. Further studies are needed to elucidate the pathways of neuroprotective effects of AA after spinal trauma.WoSScopu

Crustal flow driving twin domes exhumation and low-angle normal faulting in the Menderes Massif of western Anatolia

Lower crustal flow in regions of post-orogenic extension has been inferred to explain the exhumation of metamorphic core complexes and associated low-angle normal (detachment) fault systems. However, the origin of detachment faults, whether initially formed as high-angle or low-angle shear zones, and the extension is symmetric or asymmetric remains enigmatic. Here, we use numerical modeling constrained by geophysical and geological data to show that symmetric extension in the central Menderes Massif of western Anatolia is accommodated by the crustal flow. Our geodynamic model explains how opposite dipping Gediz and Büyük Menderes detachment faults are formed by ∼40° footwall rotation. Model predictions agree with seismic tomography data that suggests updoming of lower crust beneath the exhumed massifs, represented as “twin domes” and a flat Moho. Our work helps to account for the genetic relation between the exhumation of metamorphic core complexes and low-angle normal faulting in both Cordillera and Aegean orogenic regions and has important implications on crustal dynamics in extensional provinces

High Flow Nasal Oxygen Therapy Usage in the Adult Intensive Care Units in Turkey: Multi-center, Prospective Study

European-Respiratory-Society (ERS) International Congress -- SEP 28-OCT 02, 2019 -- Madrid, SPAINWOS: 000507372405066…European Respiratory So

Comparative Biochemical And Motor Function Analysis Of Alpha Lipoic Acid And N-Acetyl Cysteine Treatment On Rats With Experimental Spinal Cord Injury

AIM: Spinal Cord Injury (SCI) is a devastating health problem both for the patient and the clinician. Numerous treatment modalities have been studied to reverse the effects of spinal cord injury. Herein is reported the effects and the comparison of Alpha Lipoic Acid and N-Acetyl Cysteine on rats with SCI. MATERIAL and METHODS: 38 adult male Sprague-Dawley rats were randomly divided into 5 groups: only laminectomy, laminectomy and trauma, laminectomy trauma and Alpha Lipoic Acid 100 mg/kg IP administration, laminectomy trauma and N-Acetyl Cysteine 300 mg/kg IP administration, and vehicle group (PEG). The trauma model was the Modified Allen Weight drop method. After the procedure, the rats' motor function was evaluated using the modified Tarlov Scale and consequently they were sacrificed and the spinal cord tissue was analyzed biochemically for inflammation markers. RESULTS: Both Alpha Lipoic Acid and N-Acetyl Cysteine administration after the injury significantly improved the results. There was no statistically significant difference in between the agents. CONCLUSION: Although these agents both proven to be effective in ameliorating the effects of SCI, there was not enough evidence in this research to conclude the benefit of one agent over the other.WoSScopu