PLA Levels Research Articles

Propolis Reduces Inflammation and Dyslipidemia Caused by High-Cholesterol Diet in Mice by Lowering ADAM10/17 Activities.

Atherosclerosis is one of the most important causes of cardiovascular diseases. A disintegrin and metalloprotease (ADAM)10 and ADAM17 have been identified as important regulators of inflammation in recent years. Our study investigated the effect of inhibiting these enzymes with selective inhibitor and propolis on atherosclerosis. In our study, C57BL/6J mice (n = 16) were used in the control and sham groups. In contrast, ApoE-/- mice (n = 48) were used in the case, water extract of propolis (WEP), ethanolic extract of propolis (EEP), GW280264X (GW-synthetic inhibitor), and solvent (DMSO and ethanol) groups. The control group was fed a control diet, and all other groups were fed a high-cholesterol diet for 16 weeks. WEP (400 mg/kg/day), EEP (200 mg/kg/day), and GW (100 µg/kg/day) were administered intraperitoneally for the last four weeks. Animals were sacrificed, and blood, liver, aortic arch, and aortic root tissues were collected. In serum, total cholesterol (TC), triglycerides (TGs), and glucose (Glu) were measured by enzymatic colorimetric method, while interleukin-1β (IL-1β), paraoxonase-1 (PON-1), and lipoprotein-associated phospholipase-A2 (Lp-PLA2) were measured by ELISA. Tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), myeloperoxidase (MPO), interleukin-6 (IL-6), interleukin-10 (IL-10), and interleukin-12 (IL-12) levels were measured in aortic arch by ELISA and ADAM10/17 activities were measured fluorometrically. In addition, aortic root and liver tissues were examined histopathologically and immunohistochemically (ADAM10 and sortilin primary antibody). In the WEP, EEP, and GW groups compared to the case group, TC, TG, TNF-α, IL-1β, IL-6, IL-12, PLA2, MPO, ADAM10/17 activities, plaque burden, lipid accumulation, ADAM10, and sortilin levels decreased, while IL-10 and PON-1 levels increased (p < 0.003). Our study results show that propolis can effectively reduce atherosclerosis-related inflammation and dyslipidemia through ADAM10/17 inhibition.

Gold nanoparticles green-mediated by Descurainia sophia extract for the treatment of ovalbumin-induced asthma in rats

This study reveals a sustainable and eco-friendly protocol towards the in situ mediated gold nanoparticles by Descurainia sophia extract (Au NPs), followed by the assessment of its biological and chemical properties. The mediated gold NPs were characterized by FT-IR, SEM, UV–Vis, and TEM. The fabricated Au NPs were found to be in polymorphic shapes and aggregated which possibly was due to dehydration exerted during the sample preparation for SEM analysis. In addition, the particles range size was 10–38 nm. In the in vivo design, four groups of wistar rats were applied; sensitized to ovalbumin, non-sensitized, sensitized and dexamethasone-treated, and gold NPs-treated (5 μg/mL). To induce asthma, 100 mg Al(OH)3 plus 1 mg/kg ovalbumin was administered and rats were exposed to 2 % ovalbumin aerosol. 24 h after the sensitization end, rats were sacrificed. The levels of IFN-γ/IL-4 ratio, interleukin 4, IFN-γ, phospholipase A2, immunoglobulin E, total protein and lung pathological changes were determined. The Au NPs amelioration effects on pathological changes, inflammatory and immunological parameters more potent than dexamethasone offers the nanoparticle remedial property in asthma. The Au NPs significantly ameliorated the sensitized rat’s pathological changes. Treatment with gold nanoparticles resulted in reduced IgE, PLA2, TP, and IL-4 levels. A significant rise in IgE, PLA2, TP, and IL-4 concentrations, all lung pathological indices, and a notable reduction in IFN-γ/IL-4 ratio was observed.

Linking biomarkers with healthy lifestyle outcomes after stroke: Supplementary results of a 12-month randomized controlled trial

Background and aimsParticipation in a healthy lifestyle intervention such as the Diabetes Prevention Program Group Lifestyle Balance–adapted for stroke (GLB-CVA) may reduce stroke burden. Identifying biomarkers associated with lifestyle changes may enhance an individualized approach to stroke recovery. We investigated metabolic biomarkers related to cardiovascular and neurological function in individuals with stroke in the GLB-CVA study and healthy (non-stroke) individuals. Methods and resultsParticipants with chronic (>12 months) stroke were recruited to this wait-list randomized controlled trial if they were overweight (BMI ≥25 kg/m2). Participants were randomized to (1) the GLB-CVA program to complete 22 educational sessions addressing behavioral principals of dietary and physical activity or (2) a 6 month wait-list control (WLC). Biomarkers [Plasma irisin, vascular endothelial growth factor, lipoprotein-associated phospholipase A2 (Lp-PLA2), insulin-like growth factor 1 and brain-derived neurotrophic factor (BDNF)] were collected at baseline, 3, and 6 months. Age-matched healthy individuals were recruited for biomarker assessment. Compared to healthy adults (n = 19), participants with stroke (GLB-CVA = 24; WLC = 24) at baseline had higher tHcy levels (p < 0.001) and lower PLA2 levels (p = 0.016). No statistically significant interactions were observed for any biomarkers between the GLB-CVA and WLC or between people who achieved 5% weight loss and those who did not. ConclusionParticipation in a 6-month healthy lifestyle program did not result in statistically significant changes to select metabolic biomarker levels for our participants with chronic stroke. However, participants with stroke demonstrated a unique biomarker profile compared to age-matched healthy individuals.

Secretory Phospholipase A2 and Interleukin-6 Levels as Predictive Markers of the Severity and Outcome of Patients with COVID-19 Infections.

Coronavirus disease (COVID-19) has become a global pandemic. COVID-19 patients need immediate diagnosis and rehabilitation, which makes it urgent to identify new protein markers for a prognosis of the severity and outcome of the disease. The aim of this study was to analyze the levels of interleukin-6 (IL-6) and secretory phospholipase (sPLA2) in the blood of patients regarding the severity and outcome of COVID-19 infection. The study included clinical and biochemical data obtained from 158 patients with COVID-19 treated at St. Petersburg City Hospital No. 40. A detailed clinical blood test was performed on all patients, as well as an assessment of IL-6, sPLA2, aspartate aminotransferase (AST), total protein, albumin, lactate dehydrogenase (LDH), APTT, fibrinogen, procalcitonin, D-dimer, C-reactive protein (CRB), ferritin, and glomerular filtration rate (GFR) levels. It was found that the levels of PLA2, IL-6, APTV, AST, CRP, LDH, IL-6, D-dimer, and ferritin, as well as the number of neutrophils, significantly increased in patients with mild to severe COVID-19 infections. The levels of IL-6 were positively correlated with APTT; the levels of AST, LDH, CRP, D-dimer, and ferritin; and the number of neutrophils. The increase in the level of sPLA2 was positively correlated with the levels of CRP, LDH, D-dimer, and ferritin, the number of neutrophils, and APTT, and negatively correlated with the levels of GFR and lymphocytes. High levels of IL-6 and PLA2 significantly increase the risk of a severe course by 13.7 and 2.24 times, and increase the risk of death from COVID-19 infection by 14.82 and 5.32 times, respectively. We have shown that the blood levels of sPLA2 and IL-6 increase in cases which eventually result in death and when patients are transferred to the ICU (as the severity of COVID-19 infection increases), showing that IL-6 and sPLA2 can be considered as early predictors of aggravation of COVID-19 infections.

Serum lipoprotein phospholipase A2 level has diagnostic value for cognitive impairment in type II diabetes patients with white matter hyperintensity

This prospective study investigated the relationship between the lipoprotein-associated phospholipase A2 (Lp-PLA2) level and cognitive impairment (CI) in type II diabetes mellitus (TIIDM) patients with white matter hyperintensity (WMH). A total of 87 TIIDM patients diagnosed with WMH were included in this study. They were grouped into CI group and noncognitive impairment (NCI) group based on the MoCA scales. The serum Lp⁃PLA2 levels and MoCA scores of the patients with WMH were compared with those of control (Ctl). Logistic regression was used to analyze the risk factors affecting CI and diagnostic value of serum Lp⁃PLA2 for CI. The WMH group had significantly higher serum level of Lp⁃PLA2 and significantly lower MoCA score than Ctl group. There were significant differences in serum levels of homocysteine, high-density lipoprotein cholesterol (HDL-C) and Lp⁃PLA2 (P < 0.05) between the groups. Logistic regression showed that HDL-C, homocysteine and Lp⁃PLA2 were the risk factors for CI in the patients (P < 0.05); receiver operating characteristic curve analysis showed that HDL-C and Lp⁃PLA2 had significant diagnostic value for CI in WMH patients. Therefore, Lp⁃PLA2 can be assessed in the elderly as a screening to identify putative CI patents for preventive treatment and management.

Protein expression changes during phagocytosis influenced by low-frequency electromagnetic field exposure

The aim of our studies was to determine the influence of a low-frequency electromagnetic field (EMF) on the phagocytosis of latex beads (LBs) and the expression level of proteins/genes in the human monocytic macrophage Mono Mac 6 (MM6) cell line in in vitro conditions. Before phagocytosis assay cells were pre-stimulated with infectious agents such as lipopolysaccharide (LPS), Staphylococcal enterotoxin B (SEB), or the proliferatory agent phytohaemagglutinin (PHA), and then exposed to EMF (30 mT, 7 Hz, 3 h). The expression of cytoplasmic proteins like iPLA, cPLA, iNOS, NLR3/4, and Hsp70 involved in the immune response pathways to phagocytosed particles were evaluated with the usage of the Western blot analysis. mRNA encoding the iNOS protein was detected by reverse transcription PCR method. The most meaningful changes were observed for PLA2 and NLC4 proteins level and between iNOS protein expression and mRNA encoding iNOS protein amount. The EMF exposure exerted the strongest effect on iNOS encoding mRNA in cells pre-stimulated with LPS or SEB and phagocytosing LBs. The influence of EMF on phagocytosis was experimentally proved for the first time and there is a need for further investigations in term of the usage of EMF as a prospect, supportive therapy.

Aspirin resistance and blood biomarkers in predicting ischemic stroke recurrence: An exploratory study.

BACKGROUND:Recurrent strokes cause greater complications and worse outcomes by adding to the existing neurological deficit. There is the paucity of data on serum markers of inflammation as predictors of recurrent stroke. This study was planned to analyze the clinico-etiological profile of recurrent noncardioembolic ischemic stroke, estimate aspirin resistance among regular aspirin users and evaluate blood biomarkers high-sensitivity C-reactive protein (hsCRP), Tumor necrosis factor-alpha (TNF-α), Lipoprotein-associated phospholipase A2 (Lp-PLA2) as probable predictors of stroke recurrence.METHODS:Patients of recurrent noncardioembolic ischemic stroke fulfilling the inclusion criteria were enrolled. Detailed history, clinical examination, and investigations were obtained as per protocol. Aspirin resistance was determined by light transmission aggregometry. Serum hsCRP, TNF-α, and Lp-PLA2 levels were estimated.RESULTS:This study included 34 males and 16 females. Majority of the patients were > 60 years (n = 30, 60%). Thirty (60%) cases had a repeat stroke after 1 year of primary event. Thirty-nine (78%) study participants had hypertension, while 15 (30%) had diabetes. Middle cerebral artery (n = 40, 80%) was the most common vascular territory. Thirty-one (62%) cases belonged to TOAST subtype 1 (large artery atherosclerosis). Seventy two percent cases were prescribed aspirin after index stroke, but only 36% were compliant. Median (range) hsCRP level was 7.5 (0.3–155) mg/L with 72% of patients having high hsCRP level (>3 mg/L). Median (range) serum PLA2 level was 11.98 (3.31–87.24) ng/ml in patients and 6.96 (0.15–61.42) ng/ml in controls (P = 0.029). Median (range) serum TNF-α level in patients was significantly higher than controls (68.22 [1.3–287] pg/ml versus 0.098 [0.002–36.31] pg/ml, P < 0.001). Aspirin resistance was found in 41.7% patients while 16.7% were semi-resistant. Mean % platelet aggregation was 34.75 ± 21.58 in patients and 64.75 ± 16.98 for controls (P < 0.001).CONCLUSIONS:Majority of patients with recurrent stroke were elderly (>60 years), hypertensive, and non-compliant with aspirin. Aspirin resistance was an important factor in patients with antiplatelet compliance. Inflammatory biomarkers hsCRP, PLA2, and TNF-α were found to be significantly elevated in patients compared to controls.

Brain Derived Neurotrophic Factor Deficiency is Associated with Cognitive Impairment and Elevated Phospholipase A2 Activity in Plasma of Mice

Decreased levels of Brain-Derived Neurotrophic Factor (BDNF) are a common finding in schizophrenia. Another well-documented protein linked to schizophrenia is intracellular Ca2+-independent Phospholipase (PLA2). However, the potential association between PLA2 and BDNF with regard to schizophrenia has yet to be examined.In the present study, male and female BDNF knockout mice, a possible genetic model of schizophrenia, were exposed to prenatal stress and tested in the nest test, open field test and T-maze. Following behavioral tests, whole brain and plasma samples were harvested to measure the activity of PLA2. BDNF knockout mice showed cognitive deficits in the T-maze. Furthermore, there was a quadratic association of PLA2 with performance in the open field test. Moreover, BDNF deficiency and female sex were associated with elevated plasma PLA2 levels. The cognitive impairment of BDNF heterozygous mice as well as their increased PLA2 activity in plasma is consistent with findings in schizophrenia patients. The particular elevation of PLA2 activity in females may partly explain sex differences of clinical symptoms in schizophrenia (e.g. age of onset, severity of symptoms). Additionally, PLA2 was significantly correlated with body and adrenal weight after weaning, whereby the latter emphasizes the possible connection of PLA2 with steroidogenesis.

Hepatopancreas transcriptome analysis reveals the molecular responses to different dietary n-3 PUFA lipid sources in the swimming crab Portunus trituberculatus

n-3 LC-PUFA is vital to crustaceans for the growth, molting and development, while the precise molecular mechanism underlying the regulation of n-3 LC-PUFA is poorly investigated. Herein, an 8-week nutritional trial and transcriptome analysis were used to investigate the impacts of dietary n-3 PUFA lipid sources including fish oil (FO), krill oil (KO) and linseed oil (LO) on the transcriptome characteristics in hepatopancreas of swimming crab (Portunus trituberculatus). Crabs fed KO diet had significantly higher percent weight gain (PWG) and molting ratio (MR) than those of crabs fed other diets. The transcriptome analysis reveled that a total of 6567 unigenes were annotated in KEGG database, of which the expression of 1126 significantly altered in the KO vs FO group, while the expression of 921 unigenes changed in the LO vs FO group. The differentially expressed genes (DEGs) in KO vs FO comparison group involved in fat digestion and absorption, ABC transporters, adipocytokine signaling pathway, protein digestion and absorption, glycolysis/gluconeogenesis and HIF-1 signaling pathway. While dietary LO significantly altered the expression levels of genes involved in carbohydrate digestion and absorption, mineral absorption, protein processing in endoplasmic reticulum, glycosphingolipid biosynthesis and insect hormone biosynthesis. In addition, dietary KO significantly up-regulated the expression levels of cyp18a1 (participates in the process of ecdysteroids biosynthesis and further controls the molting of crustaceans), tryp (trypsin-like serine proteinases, resists the pathogen invasion), pla2 (phospholipase A2, regulates different inflammatory processes), ABC transporter (ATP-binding cassette transporter, transports amino acids, lipids and proteins into and out of the cell), cpt1 (carnitine palmitoyltransferase I, the major rate-limiting enzyme of mitochondrial fatty acid β-oxidation) and slc2 (solute carrier 2, mediates bidirectional glucose transport). Meanwhile, dietary LO could significantly down-regulated the expression level of cyp315a1 compared to dietary FO. To sum up, swimming crab fed with dietary krill oil could not only enhance the transport of glucose, lipids, fatty acids and other substrates, but also up-regulate the expression levels of genes related to molting, immunity and energy metabolism to promote molting and growth of swimming crab. The present study revealed the molecular responses of crabs to different dietary n-3 PUFA lipid sources, and further provided important clues with respect to elucidating the molecular mechanisms underlying the regulation of EPA and DHA on the molting and growth of swimming crab.

Serum concentration of phospholipase A2 and platelet aggregation during hyperbaric oxygen therapy in patients with ischemic heart disease

To study changes in serum PLA2 concentration and its relation to platelet aggregation activity during HBO in patients with stable CHD. We examined 42 patients with stable angina FC II-III, 27 received antiplatelet therapy (Cardiomagnyl 75 mg: acetylsalicylic acid + magnesium hydroxide), and 15 patients did not. All patients received a 10-day course of HBO at 1.2 atmosphere mode for 40 min. Platelet hemostasis and serum PLA2 concentration were evaluated. Platelet aggregation was tested using Biola LA-230-2 aggregation analyzer (Biola Scientific, Russia). The platelets count and mean platelet volume (MPV) were determined on a Mindray BS-3200 hematology analyzer (Mindray, China). PLA2 levels were determined by enzyme immunoassay using Model 680 microplate reader (Bio-Rad, USA). Residual platelet reactivity was evaluated by 5.0 ADP-induced aggregation. Assessment of the HBO effect on the functional state of platelets depending on their aggregation activity and the therapy taken showed a significant increase in spontaneous aggregation and ADP-induced aggregation at inducer concentration of 1.0 μM (p=0.049) in patients with baseline hyperaggregation taking Cardiomagnyl after HBO. No significant changes in PLA2 concentration were observed. At the same time, patients with baseline hyperaggregation who did not take antiplatelet agents had no changes in platelet aggregation activity and a decreased serum PLA2. In patients with baseline normal aggregation receiving an antiplatelet drug, a course of HBO had no effect on platelet aggregation activity and PLA2 level. In patients with baseline normal aggregation who did not take antiplatelet agents, a course of HBO resulted in significant decrease in PLA2 levels and no changes in platelet aggregation activity. In patients with low aggregation activity (hypoaggregation) who took antiplatelet agents, a significant increase in spontaneous aggregation and no change of serum PLA2 after an HBO course was observed. The study showed a divergent response to the hyperbaric oxygen, depending on the antiplatelet therapy and the background aggregation.

Antihypertensive activity of oleanolic acid is mediated via downregulation of secretory phospholipase A2 and fatty acid synthase in spontaneously hypertensive rats.

Oleanolic acid (OA) is reported to possess antihypertensive activity via the regulation of lipid metabolism; however, the mechanisms underlying lipid regulation by OA are yet to be fully elucidated. The aim of the present study was to evaluate the mechanisms via which OA regulates lipid metabolism in spontaneously hypertensive rats (SHRs) via ultra-performance liquid chromatography-quadrupole/Orbitrap-mass spectrometry (MS)-based lipidomics analysis. SHRs were treated with OA (1.08 mg/kg) for 4 weeks. The liver tissues were excised, homogenized in dichloromethane and centrifuged, and subsequently the supernatant layer was collected and concentrated under vacuum to dryness. The dichloromethane extract was subjected to MS analysis and database searching, and comparison of standards was performed to identify potential biomarkers. Partial least squares-discriminant analysis performed on the liver lipidome revealed a total of 14 endogenous metabolites that were significantly changed in the SHR model group (SH group) compared with Wistar Kyoto rats [normal control (NC group)], including glycerophospholipids, sphingolipids and glycerides. Heatmaps revealed that the liver lipid profiles in the OA group were clustered more closely compared with those observed in the NC group, indicating that the antihypertensive effect of OA was mediated via regulation of liver lipid metabolites. It was observed that the protein levels of secretory phospholipase A2 (sPLA2) and fatty acid synthase (FAS) were increased in the SH group compared with the NC group. In addition, the levels of lysophosphatidylcholine and triglycerides in the liver were elevated, whereas the levels of low-density lipoprotein cholesterol and high-density lipoprotein cholesterol were reduced in the SH group. Upon treatment with OA, the mRNA and protein levels of PLA2 and FAS were observed to be downregulated. Collectively, the present study indicated that the antihypertensive activity of OA was mediated via downregulation of sPLA2 and FAS in SHRs, and that treatment with OA resulted in significant improvements in blood pressure and associated abnormalities in the lipid metabolites.

Altered Metabolism of Phospholipases, Diacylglycerols, Endocannabinoids, and N-Acylethanolamines in Patients with Mastocytosis

Background Mastocytosis is a condition characterized by the expansion and accumulation of mast cells (MCs) in various organs. The symptoms are related to the increased release of MC-derived mediators that exert local and distant effects. MCs are a source and target of phospholipase enzymes (PLs), which catalyze the cleavage of membrane phospholipids releasing lipid mediators (e.g., diacylglycerols (DAGs) and the endocannabinoid (EC) 2-arachidonoylglycerol (2-AG)). To date, there are no data on the role of these lipid mediators in mastocytosis. Here, we analyzed plasma levels of PLA2, PLC, DAG, ECs, and EC-related N-acylethanolamines in patients with mastocytosis. Methods In 23 patients with mastocytosis and 23 healthy individuals, we measured plasma PLA2 and PLC activities, DAG, 2-AG, anandamide (AEA), palmitoylethanolamide (PEA), and oleoylethanolamide (OEA). Results Plasma PLA2 and PLC activities were increased in mastocytosis patients compared to controls. Concentrations of DAG (18:1 20:4 and 18:0 20:4), two second messengers produced by PLC, were higher in mastocytosis compared to controls, whereas the concentrations of their metabolite, 2-AG, were not altered. AEA was decreased in mastocytosis patients compared to controls; by contrast, AEA congener, PEA, was increased. PLA2 and PLC activities were increased only in patients with mediator-related symptoms. Moreover, PLC activity was positively correlated with disease severity and tryptase concentrations. By contrast, AEA was negatively correlated with tryptase concentrations. Conclusions PLs and some lipid mediators are altered in patients with mastocytosis. Our results may pave the way for investigating the functions of these mediators in the pathophysiology of mastocytosis and provide new potential biomarkers and therapeutic targets.

Rosmarinic acid affects immunological and inflammatory mediator levels and restores lung pathological features in asthmatic rats

BackgroundThe effects of rosmarinic acid (RA) on immunological and inflammatory mediator levels in bronchoalveolar lavage fluid (BALF) as well as lung pathological changes in asthmatic rats were investigated. MethodsThe levels of IFN-γ, IL-4, IFN-γ/IL-4 ratio, IgE, PLA2, and total protein (TP) in BALF and pathological changes in the lung were evaluated in control group (C), asthma group (sensitized to ovalbumin) (A), asthma groups treated with RA and dexamethasone. ResultsCompared to the control group, asthmatic rats showed increased levels of IL-4, IgE, PLA2, and TP as well as all pathological scores with decreased levels of IFN-γ and IFN-γ/IL-4 ratio (P<0.05 to P<0.001). The levels of IL-4, IgE, PLA2, and TP significantly reduced in groups treated with all concentrations of RA compared to asthma group (P<0.001 for all cases). IFN-γ was significantly decreased in groups treated with two lower concentrations of RA but IFN-γ/IL-4 ratio was increased in groups treated with two higher concentrations of RA compared to asthma group (P<0.05 to P<0.001). Treatment with all doses of RA led to significant improvement in pathological scores in asthmatic animals (P<0.05 to P<0.001). Most measured parameters were also significantly improved in dexamethasone-treated animals (P<0.01 to P<0.001) but IFN-γ/IL-4 ratio and the scores of interstitial fibrosis, bleeding and epithelial damage did not change in this group. ConclusionThe results indicated a preventive effect for RA on immunological and inflammatory mediators as well as lung pathological changes in asthmatic rats which were comparable or even more potent than that of dexamethasone.

Role of the placenta in serum autotaxin elevation during maternal cholestasis.

Intrahepatic cholestasis of pregnancy (ICP) is frequently accompanied by pruritus, whose etiology has been associated with an enhanced production of lysophosphatidic acid (LPA) by the combined action of phospholipase A1/A2 (PLA1/PLA2) and autotaxin (ATX). Here, we have investigated whether the placenta is involved in LPA release to maternal circulation during ICP. Serum levels of ATX and LPA (determined by ELISA) were elevated in women with ICP, and a correlation between both parameters was found. No relationship between serum levels of ATX or LPA and bile acids was found. Expression levels of ATX and PLA2 were determined by RT-qPCR and Western blot. Placenta ATX but not PLA2 was significantly upregulated in ICP, and a tendency to increase was found at the protein level. A correlation between serum ATX and placental ATX mRNA levels was found. In human placenta at term, ATX was clearly detected (by immunofluorescence) in Hofbauer cells, but only faintly in trophoblast cells. In pregnant rats, the expression of Atx and Pla2 in placenta was lower than in liver. When obstructive cholestasis was imposed by bile duct ligation from day 14 of gestation until term, placenta Atx and Pla2 expression was markedly enhanced, and overexpression was confirmed at the protein level for Pla2, whereas Atx protein was not detected. In conclusion, the placenta substantially participates in LPA production during gestation. This contribution is markedly higher during maternal cholestasis and hence, may be involved in ICP-associated pruritus. NEW & NOTEWORTHY Fetal placental macrophages and, to a lesser extent, trophoblast cells express high levels of autotaxin at term. An increased expression of mRNA and protein autotaxin, the key secretory enzyme responsible for the production of lysophosphatidic acid in serum, has been observed in placentas of women with cholestasis of pregnancy, which supports that the placenta can contribute to an increased production of this pruritogenic compound in women suffering from this liver disease.

Expression of factors involved in apoptosis and cell survival is correlated with enzymes synthesizing lysophosphatidic acid and its receptors in granulosa cells originating from different types of bovine ovarian follicles

BackgroundLysophosphatidic acid (LPA) regulates reproductive processes in the cow. Ovarian granulosa cells play a pivotal role in follicle growth and development. Nevertheless, the role of LPA in the local regulation of granulosa cell function in different follicle categories in the bovine ovary has not been investigated.MethodsOvarian follicles were divided into healthy, transitional and atretic categories. The expression levels of AX, PLA2, LPARs and factors involved in apoptosis and cell survival processes in granulosa cells in different types of follicles were measured by real-time PCR. The correlations between the expression levels of AX, PLA2, LPARs and the examined factors were measured. The immunolocalization of AX, PLA2 and LPARs in different ovarian follicles was examined by immunohistochemistry. Statistical analyses were conducted in GraphPad using a one-way ANOVA followed by the Kruskal-Wallis multiple comparison test or a correlation analysis followed by Pearson’s test.ResultsThe expression levels of AX, PLA2 and LPARs, with the major role of LPAR2 and PLA2, were found in the granulosa cells originating from different follicle types. The expression levels of the factors involved in cell apoptosis (TNFα and its receptors, FAS, FASL, CASP3, CASP8, β-glycan, and DRAK2) were significantly higher in the granulosa cells of the atretic follicles compared to the healthy follicles. A number of correlations between LPARs, AX, PLA2 and factors associated with apoptosis were observed in the atretic but not in the healthy follicles. A greater expression of the factors involved in differentiation and proliferation in the granulosa cells (DICE1 and SOX2) was found in the healthy follicles in comparison with the atretic. A number of correlations between LPARs, AX, PLA2 and the factors associated with cell survival were observed in the healthy but not in the atretic follicles.ConclusionsGranulosa cells are the target of LPA action and the source of LPA synthesis in the bovine ovarian follicle. We suggest that the participation of LPA in apoptosis in the atretic follicles mainly occurs through the regulation of TNF-α-dependent and caspase-induced pathways. In the transitional follicles, LPA might influence the inhibins to shift the balance between the number of healthy and atretic follicles. In the healthy follicle type, LPA, acting via LPAR1, might regulate MCL1 and estradiol-stimulating ERβ mRNA expression, leading to the stimulation of anti-apoptotic processes in the granulosa cells and their differentiation and proliferation.

Anti‐inflammatory, antioxidant, and immunomodulatory effects of curcumin in ovalbumin‐sensitized rat

Anti-inflammatory and antioxidant properties of curcumin have been shown. In this study, anti-inflammatory, antioxidant, and immunomodulatory effects of curcumin in sensitized rat were evaluated. Six groups of rats including control (C), ovalbumin-sensitized (as a rat model of asthma, S), S groups treated with curcumin (Cu 0.15, 0.30, and 0.60 mg/mL), and 1.25 μg/mL dexamethasone (S + D) were studied. Curcumin and dexamethasone were given in animals' drinking water during sensitization period. Total and differential WBC count, PLA2, TP, IFN-γ, IL-4, IgE, oxidant, and antioxidant biomarker levels in bronchoalveolar lavage fluid (BALF) were examined. Total WBC, neutrophil and eosinophil counts, levels of PLA2, TP, IgE, IL-4, NO2 , NO3 , and MDA in BALF were increased but lymphocyte percentage, SOD, CAT, thiol, and IFN-γ levels and IFN-γ/IL-4 ratio decreased in S animals compared to controls (P < 0.001 for all cases). Treatment with all concentrations of curcumin significantly improved total WBC, PLA2, TP, IgE, IL-4, IFN-γ, IFN-γ/IL-4 ratio, SOD, thiol, NO2 , and NO3 compared to S group (P < 0.01 to P < 0.001). Two higher concentrations of curcumin significantly decreased neutrophil and eosinophil counts and MDA level but increased IFN-γ, CAT and lymphocyte values compared to S group (P < 0.001 for all cases). Dexamethasone treatment also significantly improved most of the measured parameters (P < 0.05 to P < 0.001) but it did not change IL-4 and IFN-γ levels and IFN-γ/IL-4 ratio. Anti-inflammatory, antioxidant, and immunomodulatory effects of curcumin with more specific immunomodulatory effect on Th1/Th2 balance compared to dexamethasone in sensitized rats was shown. © 2017 BioFactors, 43(4):567-576, 2017.

Glasgow Coma Scale Improvement after Lidocaine Infusion in Moderate Traumatic Brain Injury

Since the Glasgow Coma Scale was developed 40 years ago it has been accepted throughout the world as a method for assessing impaired consciousness. Fourty moderate TBI patients were simple randomly divided into 2 groups: group A take normal saline infusion without lidocaine as control group (N=20), and group B take normal saline with lidocaine at rate 1 mg/kg/BW infusion as treated groups (N=20). Serum samples were collected at before induction and 2 hours after giving infusion treatment to measure serum IL-6 and PLA2 level, and lidocaine serum concentration. GCS were measured before induction and after 24 hours surgery. Data were analyzed using analytic statistic with paired t test to measure serum IL-6, PLA2, & lidocaine serum level before induction and after 2 hours. The correlation of serum IL-6, PLA2 and GCS was measure with Separman correlation. A P value less than 0.05 were considered significant. The result are both serum IL-6 and PLA2 levels were significantly lower in saline with lidocain infusion group (p<0.005) with Mann Whitney and Dependent t test statistically analyses. The lower serum IL-6 and PLA2 levels, the higher GCS value. GCS measurement was significantly higher in lidocaine infusion group (p<0.05) with dependent t test. The decrese in serum IL-6 and PLA2 was correlated with the dosage of lidocaine. Medium doses of lidocaine achieved the optimum decreases in the serum IL 6 and PLA2 level. Therefore, inhibition of the composition and secretion of IL-6 and PLA2 levels by Lidocaine might be one of the mechanisms involved in decreasing inflammatory reaction and cellular damage in brain cells, and may be responsible for the neuroprotective effect after TBI. Glasgow Coma Scale was improving after infusion with lidocaine 1 mg/ kg/ hr during traumatic brain injury surgery by decreasing IL-6 and PLA2 plasm.

Plasma secreted phospholipase A2 in asthmatic children: correlation with leptin levels and exercise induced bronchoconstriction

Purpose: Dysregulated cysteinyl leukotriene (CysLT) synthesis is prominent in exercise-induced bronchoconstriction (EIB). Secreted phospholipase A2 (sPLA2) plays a key regulatory role in the biosynthesis of CysLTs. We previously found that serum leptin levels correlate with (EIB) in children with asthma. The aim of this study was to address the relationship between plasma sPLA2/leptin levels and EIB. Methods: Sixty-seven prepubertal children between the ages of 6 and 10 years were included in the study. They were asthmatics with EIB (n= 25), asthmatics without EIB (n= 21), and healthy subjects (n= 21). We measured the plasma sPLA2 and leptin levels. We also performed pulmonary function tests at baseline, after bronchodilator inhalation, and after exercise. Results: The sPLA2 and leptin levels were significantly higher in asthmatics with EIB than in those without and control subjects. In addition, sPLA2 levels were significantly correlated with body mass index (Speraman correlation coefficient r= 0.343, P= 0.023) and leptin levels (partial correlation coefficient r= 318, P= 0.033). The maximum decrease in % forced expiratory volume in 1 second after exercise was significantly correlated with both PLA2 levels (r= 0.301, P= 0.041) and leptin levels (r= 0.346, P= 0.018). Conclusion: The sPLA2 and leptin levels were significantly higher in asthmatics with EIB than in asthmatics without EIB and control subjects. In addition, sPLA2 levels were significantly correlated with leptin levels and EIB in asthmatic children. (Allergy Asthma Respir Dis 2015;3:99-104)

Non-steroidal anti-inflammatory drugs attenuate the vascular responses in aging metabolic syndrome rats.

Metabolic syndrome (MS) and aging are low-grade systemic inflammatory conditions, and inflammation is a key component of endothelial dysfunction. The aim of this study was to investigate the effects of non-steroidal anti-inflammatory drugs (NSAIDs) upon the vascular reactivity in aging MS rats. MS was induced in young male rats by adding 30% sucrose in drinking water over 6, 12, and 18 months. When the treatment was finished, the blood samples were collected, and aortas were dissected out. The expression of COX isoenzymes and PLA2 in the aortas was analyzed using Western blot analysis. The contractile responses of aortic rings to norepinephrine (1 μmol/L) were measured in the presence or absence of different NSAIDs (10 μmol/L for each). Serum levels of pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β) in control rats were remained stable during the aging process, whereas serum IL-6 in MS rats were significantly increased at 12 and 18 months. The levels of COX isoenzyme and PLA2 in aortas from control rats increased with the aging, whereas those in aortas from MS rats were irregularly increased with the highest levels at 6 months. Pretreatment with acetylsalicylic acid (a COX-1 preferential inhibitor), indomethacin (a non-selective COX inhibitor) or meloxicam (a COX-2 preferential inhibitor) decreased NE-induced contractions of aortic rings from MS rats at all the ages, with meloxicam being the most potent. Acetylsalicylic acid also significantly reduced the maximum responses of ACh-induced vasorelaxation of aortic rings from MS rats, but indomethacin and meloxicam had no effect. NSAIDs can directly affect vascular responses in aging MS rats. Understanding the effects of NSAIDs on blood vessels may improve the treatment of cardiovascular diseases and MS in the elders.

The extract of Zataria multiflora affect tracheal responsiveness, serum levels of NO, nitrite, PLA2, TP and histamine in sensitized Guinea pigs

Ethnopharmacological relevanceDifferent therapeutic effects for Zataria multiflora have been described in Iranian traditional medicine. In addition anti-inflammatory and anti-oxidant effects of this plant were previously described. The effect of the extract of Zataria multiflora on tracheal responsiveness (TR) and inflammatory mediators of sensitized guinea pigs was examined. Materials and methodsFive groups of sensitized guinea pigs to ovalbumin (OA) were given drinking water alone, drinking water containing three concentrations of the extract and dexamethasone. TR to methacholine and OA, serum levels of NO, nitrite, PLA2, TP and histamine were measured (n=6, for each groups). ResultsTR to methacholine and OA, serum levels of NO, nitrite, PLA2, TP and histamine were increased in sensitized animals compared to control group (p<0.05 to p<0.001). Treatment of sensitized animals with dexamethasone and most concentration of the extract lead to significantly decrease in all measured parameters compared to sensitized group (p<0.05 to p<0.001) except TP and histamine levels in treated group with dexamethasone. ConclusionThese results showed a preventive effect of the extract of Zataria multiflora on tracheal responsiveness, serum level of NO, nitrite, total protein, PLA2 and histamine in sensitized guinea pigs which was equal or even more potent than dexamethasone at used concentrations.