M1 Decrease Research Articles

Abstract 14672: Regulation of Succinate Signaling and Immunometabolism in Diabetes-Associated Atherosclerosis by Microrna-369-3p

Introduction: The risk of atherosclerotic cardiovascular disease (ASCVD) is increased in people with diabetes. Diabetes (D) is associated with impaired metabolism and functionality of macrophages (Mps), resulting in chronic inflammation. Identifying factors to combat metabolic alterations and promote resolution of inflammation remains an unmet goal. microRNAs (miRs) orchestrate multiple signaling events in Mps and regulate inflammation. Hypothesis: miR-369-3p regulates metabolic reprogramming in Mps during diabetes-associated atherosclerosis (D-AS). Methods and Results: miRNA-seq profiling from aortic intimal lesions of LDLR-/- mice on a high-fat sucrose containing (HFSC) diet revealed miR-369-3p downregulation with D-AS progression. miR-369-3p was also downregulated in PBMCs derived from D and D-associated ASCVD patients. Cell-type profiling showed miR-369-3p enrichment in mouse bone marrow derived Mps (BMDMs). oxLDL-stimulated BMDMs (foam cells, FCs) resulted in marked downregulation of miR-369-3p, increased secretion of IL1-β, and metabolic shift of oxidative phosphorylation (OXPHOS) to glycolysis, while miR-369-3p overexpression (miR-369-3p O/E) favored OXPHOS over glycolysis and reduced IL1-β secretion. Metabolomics revealed succinate, a key immunometabolite was profoundly elevated extracellularly from FCs, while miR-369-3p O/E reversed this phenotype. Extracellular succinate activates its receptor, GPR91. Indeed, GPR91 was upregulated in FCs while miR-369-3p O/E significantly repressed GPR91. 3’UTR luciferase reporter assay confirmed GPR91 as a miR-369-3p target. miR-369-3p O/E dampened GPR91-mediated inflammasome activation and mitochondrial dysfunction in FCs by decreasing p-ERK, p-DRP1, and NLRP3. Therapeutic delivery of miR-369-3p in LDLR-/- mice fed HFSC diet for 12 weeks reduced plaque size by ORO staining and plasma IL1-β. Flow cytometry of aortic Mps revealed that miR-369-3p decreased GPR91 expression by ~50%, along with a decrease in M1 and increase in M2-like-Mps. Conclusions: miR-369-3p confers a pro-resolving role in regulating lipid-driven inflammation and metabolic rewiring in Mps via targeting GPR91 and preventing D-AS plaque progression.

Macrophages of the M1 and M2 types play a role in keloids pathogenesis.

The pathophysiology of keloid formation is unknown, however, macrophages are thought to play a role in keloid formation. Understanding the mechanism(s) of keloid development might be crucial in developing a new treatment regimen for keloids. The aim of this study was to understand possible status of M1 and M2 type macrophages in the pathogenesis of keloid. Thirty cases of Keloid tissues were selected according to our inclusion and exclusion criteria, as well as 30 normal scars, were enrolled in our study as a control group. An excisional biopsy was harvested and ELISA was done on keloid tissue and normal scar samples, with CD68, the surface marker for M1 and CD163 representing M2. The results revealed the low expression of M1 (CD68) in keloid tissue meanwhile high levels of M1 were detected in normal scars. We also detected that higher tissue expression of M2 (CD163) was significantly associated with keloid cases when compared to low M2 expression in the control group. An important finding that was discovered during our study is that the M1 and M2 are significant predictors of keloid. Every increase of 1 ng/mL in M1 decreases the risk of keloid by 0.99 while every increase of one unit in M2 increases the risk of keloid by 2.01. This study concluded that the keloid formation could be a result of an abnormal response to tissue injury where there is an excessive entry of inflammatory cells into the wound, including macrophages and that the keloid incidence might be related to a decrease in M1 and an increase in M2.

Production of NOS2 and inflammatory cytokines is reduced by selected protein kinase inhibitors with partial repolarization of HL-60 derived and human blood macrophages

JAK/STAT pathway plays a well-known role in macrophage polarization, but other signaling routes may also be involved. The aim of this study was to identify new signaling pathways and repolarize macrophages by selected protein kinase inhibitors. HL-60 derived macrophages were chosen as model cells and human blood macrophages were used for comparison. M1 and M2 polarization of HL60 derived and human blood macrophages was promoted by LPS + IFNγ (LIF) and IL-4 treatments, respectively. In HL-60 derived macrophages, M1 polarization was mediated by Erk1/2 and p38 phosphorylation, while HSP27 phosphorylation was involved in M2 polarization. The inhibition of both MAPK and JAK/STAT pathways reduced the expression of NOS2, IP-10 and TNFα, IL-8 production was decreased by the inhibition of AMPK and PKD, the upstream kinase of HSP27. HSP27 phosphorylation was inhibited by NB 142, a PKD inhibitor. The expression of CD80 (M1 marker) was reduced by MAPK and JAK/STAT inhibitors, without increasing CD206 (M2 marker). On the other hand, CD206 was reduced by PKD and AMPK inhibitors, without increasing CD80 marker. Phagocytic capacity of HL-60 derived macrophages was higher in M1 macrophages and decreased by trametinib and a p38 inhibitor, while in human blood macrophages, where AT 9283, a JAK/STAT inhibitor also caused a significant decrease in M1 polarized macrophages, no difference was observed between M1 and M2 macrophages. Our results suggest that the repolarization of macrophages cannot be achieved by inhibiting their signaling pathways; nevertheless, the expression of certain polarization markers was decreased, therefore a “depolarization” could be observed both in M1 and M2 polarized cells. Selected protein kinase inhibitors of M1 polarization, decreasing NOS 2 and inflammatory cytokines may be potential candidates for therapeutical trials against inflammatory diseases.

Color vision assessment following cataract surgery using anomaloscope

Aim: To evaluate the impact of cataract on color vision detection using an anomaloscope.
 Methods: 12 patients aged 64–82 years with history of cataract surgery in one eye and mature cataracts in the other eye were examined for color vision using the Rayleigh and the Moreland anomaloscope tests.
 Results: Different patterns of blue and green color vision deterioration have been observed in the eye with cataract as compared to the IOL eye. A decrease in M1 to approximately 1 (the minimum of the Moreland test scale) was observed in 5 out of 12 participants, whereas an increase in M2 to 99 (the maximum of the Moreland test scale) was observed in 6 out of 12 subjects. R1 found in the Rayleigh test was significantly lower in the cataract eyes as compared to the IOL eyes (38.9 ± 2.3 vs. 43.4 ± 1.6; p < 0.001).
 Conclusions: It has been shown that cataract causes a decrease in color sensitivity, especially affecting green and blue colors. Following IOL implantation, the previously deteriorated vision of green and blue was clearly improved. The most valuable parameters of the anomaloscope test, which enable to determine cataract severity, were M1, M2 and R1.

Abstract P217: Imbalance Of M1/M2 Macrophages In Ovaries Of Hypertensive Mice Is Associated With Reproductive Dysfunction And Lymphangiogenesis

Macrophages are the principal immune cells in ovaries. Besides protecting against invading pathogens and antigens, macrophages also play an essential role in folliculogenesis, ovulation, luteinization, lymphangiogenesis, and other functions. An imbalance in M1/M2 macrophages is observed in systemic circulation in patients and animals with hypertension (HTN). Although studies have demonstrated an association between HTN and reproductive dysfunction in females, the effect of HTN on ovarian M1/M2 ratio and lymphatics is largely unknown. We hypothesized that L-NAME-induced HTN (LHTN) and salt-sensitive hypertension (SSHTN) may increase the M1/M2 balance in ovaries which is associated with lymphangiogenesis and reproductive dysfunction in mice. Female mice were either provided L-NAME (0.5 mg/mL) in their drinking water for 3 weeks or L-NAME for 2 weeks, followed by a 2-week washout period and subsequent 3-week 4% high salt diet (SSHTN). Control mice (C) received tap water and normal diet. Flow cytometry analysis revealed a significant decrease in M1 (C: 46%±1, LHTN: 33%±2; p<0.05) and an increase in M2 (C: 7%±1, LHTN: 12%±1; p<0.05) macrophages in ovaries from LHTN mice. In SSHTN mice, ovaries had significantly increased M1 (C: 24%±1, SSHTN: 44%±2; p<0.05) and decreased M2 (C: 12%±1, SSHTN: 4%±1; p<0.05) macrophages. There was a significant increase in gene expression of the hormone receptors AR, FSHR, ERa, ERb, and LHR, and the steroidogenic enzymes StAR, 3bHSD, CYP11a1, and CYP17a1. Ovaries of hypertensive mice had a significant increase in gene expression of the lymphatic vessel markers Lyve-1, Podoplanin, and Prox-1, the lymphangiogenic growth factor VEGF-C and receptors VEGFR-2 and VEGFR-3. Taken together these data demonstrate that HTN disturbs M1/M2 macrophages in ovaries and is associated with reproductive dysfunction and lymphangiogenesis. Manipulation of M1/M2 ratio may be a therapeutic opportunity to improve reproductive health in hypertensive women.

Differentiating tic electrophysiology from voluntary movement in the human thalamocortical circuit

ObjectivesTourette syndrome is a neurodevelopmental disorder commonly associated with involuntary movements, or tics. We currently lack an ideal animal model for Tourette syndrome. In humans, clinical manifestation of tics cannot...

Effect of Fluconazole on the Pharmacokinetic Properties of Imrecoxib, a Novel NSAID: A Single-center, Open-label, Self-controlled Study in Healthy Chinese Male Volunteers

PurposeImrecoxib is one type of cyclooxygenase-2 inhibitor with the capability of reducing the potential cardiovascular risk caused by other NSAIDs. Co-administration with other medications can affect the cytochrome P450 (CYP) 2C9 enzyme function; thus, imrecoxib metabolism can be affected. The purpose of this research was to evaluate the effects of fluconazole, which is known to inhibit CYP2C9, on imrecoxib's pharmacokinetic (PK) parameters. MethodsIn this single-center, single-arm, open-label, self-controlled study, 12 healthy Chinese male volunteers (mean [SD] age, 22.6 [2.43] years) received the following 2 treatments separated by a washout period of 8 days under a fasting state: (1) a single oral dose of imrecoxib 100 mg; and (2) fluconazole 200 mg/d over 6 days followed by concurrent dosing of imrecoxib 100 mg and fluconazole 200 mg. Plasma concentrations of imrecoxib (M0) and its metabolites (4′-hydroxymethyl metabolite [M1] and 4′-carboxylic acid metabolite [M2]) for PK analysis were obtained at 0 (baseline) and 0.5, 1, 1.5, 2, 3, 4, 8, 12, 24, 48, and 72 hours after imrecoxib dosing. Safety and tolerability assessments were performed throughout the study. FindingsAll subjects completed the study. There was 1 adverse event; drug-induced liver damage in 1 subject occurred after he received imrecoxib plus fluconazole, and the subject recovered without any sequelae. Coadministration with fluconazole resulted in much higher plasma imrecoxib concentrations, with an increase of 88% in Cmax and 72% in AUC0–t compared with only imrecoxib treatment, which showed that fluconazole may increase plasma exposure to imrecoxib. Fluconazole also caused a small, but not clinically relevant, decrease in M1 and M2 mean Cmax (13% and 14%, respectively), but there was minimal change in M1 and M2 mean AUC0–t (3% and 2%). However, there were no statistically significant differences in vital signs, clinical laboratory test results, ECGs, or adverse events between treatments. ImplicationsConcurrent administration of imrecoxib and fluconazole did not seem to change imrecoxib's safety profile. The ratio (imrecoxib + fluconazole/imrecoxib) for AUC0–t was 1.72 (90% CI, 1.41–2.11) and for Cmax it was 1.88 (90% CI, 1.59–2.21). Hence, it is necessary to adjust the imrecoxib dose when it is concurrently used with other CYP2C9 inhibitors.

Penetrative Bénard-Marangoni Convection in a Micropolar Ferrofluid Layer via Internal Heating and Submitted to a Robin Thermal Boundary Conditions

Penetrative Benard-Maranagoni convection in micropolar ferromagnetic fluid layer in the presence of a uniform vertical magnetic field has been investigated via internal heating model. The lower boundary is considered to be rigid at constant temperature, while the upper boundary free open to the atmosphere is flat and subject to a convective surface boundary condition. The resulting eigenvalue problem is solved numerically by Galerkin method. The stability of the system is found to be dependent on the dimensionless internal heat source strength Ns, magnetic parameter M1, the non-linearity of magnetization parameter M3, coupling parameter N1, spin diffusion parameter N3 and micropolar heat conduction parameter N5. The results show that the onset of ferroconvection is delayed with an increase in N1 and N5 but hastens the onset of ferroconvection with an increase in M1, M3, N3 and Ns. The dimension of ferroconvection cells increases when there is an increase in M3, N1, N5 and Ns and decrease in M1 and N3.

Paradoxical Attenuation of Autoimmune Hepatitis by Oral Isoniazid in Wild-Type andN-Acetyltransferase–Deficient Mice

Isoniazid (INH) treatment can cause serious liver injury and autoimmunity. There are now several lines of evidence that INH-induced liver injury is immune mediated, but this type of liver injury has not been reproduced in animals, possibly because immune tolerance is the dominant response of the liver. In this study, we immunized mice with isonicotinic acid (INA)-modified proteins and Freund's adjuvant, which led to mild experimental autoimmune hepatitis (EAH) with an increase in cells staining positive for F4/80, CD11b, CD8, CD4, CD45R, and KI67. We expected that subsequent treatment of mice with oral INH would lead to more serious immune-mediated liver injury, but paradoxically it markedly attenuated the EAH caused by immunization with INA-modified hepatic proteins. In addition, patients of the slow acetylator phenotype are at increased risk of INH-induced liver injury. Treatment of arylamine N-acetyltransferase-deficient Nat1/2(-/-) mice with INH for up to 5 weeks produced mild increases in glutamate and sorbitol dehydrogenase activities, but not severe liver injury. Female Nat1/2(-/-) mice treated with INH for 1, 3, or 7 days developed steatosis, an increase in Oil Red O staining, and abnormal mitochondrial morphology in the liver. A decrease in M1 and an increase in M2a and M2b macrophages was observed in female Nat1/2(-/-) mice treated with INH for 1, 3, or 7 days; these changes returned to baseline levels by day 35. These data indicate that INH has immunosuppressive effects, even though it is also known to induce autoantibody production and a lupus-like autoimmune syndrome in humans.

Outer-diameter narrowing of the internal carotid and middle cerebral arteries in moyamoya disease detected on 3D constructive interference in steady-state MR image: is arterial constrictive remodeling a major pathogenesis?

To obtain information on affected vessels in moyamoya disease (MMD), we analyzed the vascular morphological characteristics of MMD using three-dimensional (3D) constructive interference in steady-state (CISS) magnetic resonance imaging (MRI). The population of this 3D-CISS MRI study consisted of 51 patients with MMD: 16 patients with atherosclerotic middle cerebral artery (MCA) stenosis or occlusion, 42 MRI control patients, and 28 control digital subtraction angiography (DSA) patients. We measured the outer diameters of the terminal portion of the internal carotid artery (ICA) and the proximal portion of the MCA (M1 portion). We evaluated the inner diameter as the relative value (%) obtained from magnified DSA images and analyzed these data. The outer diameters of the ICA and M1 portions were significantly smaller in the MMD group than in the other two groups, while the M1 outer diameter of the atherosclerosis group was not significantly different compared to the control (ICA: MMD, 2.61 ± 0.46 mm vs. control, 4.04 ± 0.50 mm and M1: MMD, 1.92 ± 0.43 mm vs. control, 3.34 ± 0.54 mm vs. atherosclerosis, 3.45 ± 0.56 mm). Furthermore, in MMD patients, the outer diameter was unrelated to the progression of the luminal stenosis grade estimated by DSA. This is the first report that the outer diameters of both the ICA and M1 decrease in MMD patients. Our findings suggest that the vascular constrictive changes of the affected arteries are an important phenomenon reflecting MMD pathology.

Lesions of the Cerebellar Nodulus and Uvula Impair Downward Pursuit

We studied sinusoidal (SIN) and step-ramp (SR) pursuit in two rhesus monkeys, before and after surgical lesions of the cerebellar nodulus and uvula (Nod/Uv). Eye movements were recorded using the magnetic field scleral search coil method. Pursuit targets were generated by an LCD projector and back-projected onto a tangent screen in an otherwise dark room. After the Nod/Uv lesions, both monkeys showed a reduced eye velocity during downward pursuit (SIN: 42% decrease in M1, 91% decrease in M2; SR: 37% decrease in M1, 85% decrease in M2). For SR, the decrease was seen only for the closed-loop response; initial eye acceleration did not change (P>0.05). Upward pursuit gains increased for SIN (M1: 9%, M2: 11%); they decreased for SR (M1: 27%, M2: 18%), but to a lesser degree than for downward pursuit. Horizontal pursuit was little changed in M1 but was reduced in one direction in M2, the animal with the larger lesion. The deficit in downward tracking was limited to foveal pursuit; ocular following of random-dot stimuli was retained, even when the target subtended only several degrees. Our findings support a critical role for the Nod/Uv in vertical pursuit, particularly for sustained downward pursuit. Finally, in both monkeys, the lesion increased spontaneous upward ocular drift in the dark (mean prelesion, 1.43 degrees/s; postlesion, 5.92 degrees/s), suggesting a role for the Nod/Uv in holding the eyes still and in the genesis of downbeat nystagmus.

Distribution of muscarinic receptor subtypes in rat brain from postnatal to old age

In an effort to understand the developmental changes in the distribution of muscarinic receptor subtypes (m1–m5), specific brain regions from juvenile (16-day-old), young (21-day-old) and adult (91-day-old) rats were analyzed using subtype-selective antibodies. These studies revealed significant age-dependent changes in the four brain regions examined. In cortex, and area associated with higher cognitive functions, significant increases of m2 and m4 receptors occurred between juvenile and adult rats. In the striatum, the level of m4 receptor increased with age whereas the m1, m2 and m3 receptors had reached mature levels within the first 16 days. Small but significant changes occured in the cerebellum with a decrease in m1, m3 and m4 receptor subtypes. In contrast to other brain regions, the hippocampus displayed consistent expression levels of muscarinic receptor subtypes. Suggesting that this brain region, which is involved in the foundation of numerous neural networks, requires a full complement of muscarinic receptors at a very early age. Muscarinic receptors have been shown to be important in a number of behavioral activities, including learning and memory. The changes observed in the age-dependent expression of these receptors most likely play an important role in how acetylcholine produces its effects in vivo.

Behavioural, biochemical and histological effects of trimethyltin (TMT) induced brain damage in the rat.

To assess the nature and extent of behavioural, biochemical and histological changes induced by trimethyltin (TMT), rats were treated with a single injection of TMT over a dose range of 6, 7 and 8 mg/kg i.p. Behavioural observations were performed at a minimum of 21 days after the administration of TMT. The behavioural consequences of TMT were hyperactivity in the open-field test, increased locomotor activity and deficits in passive and active avoidance behaviour, T-maze alternation and Morris Water Maze behaviour. The behavioural changes were dose dependent and were accompanied by a degree of pathological damage to the hippocampal pyramidal cells which was particularly apparent at the highest dose. The main biochemical effects of TMT involved deficits in the serotonergic and GABA-ergic systems and a decrease in M1 and M2 binding sites in the hippocampus. These results suggest that the toxic interaction of TMT with the hippocampus and other limbic brain regions may be responsible for its effect on learning and memory.

Orthopyroxenes from granulite rocks of the Wilson Terrane (Victoria Land, Antarctica): crystal chemistry and cooling history

The compositions of 17 orthopyroxenes from granulites facies rocks of the Wilson Terrane (Ross Orogen), determined by microprobe analysis, are consistent with a metamorphic origin. Three groups may be distinguished : a) orthopyroxenes with high Al 2 O 3 content (2.85-5.33 weight%) and X Mg =0.46-0.60, from garnetcordierite granulites; b) orthopyroxenes with low Al 2 O 3 (less than 0.9 wt%) and X Mg =0.49-0.62, from enderbites; c) orthopyroxenes with very low Al 2 O 3 (less than 0.4 wt%) and X Mg =0.70, from two-pyroxene mafic granulites. These differences do not reflect different metamorphic conditions, but are due to local mineral assemblages. Zoning occurs in few orthopyroxenes from garnet-cordierite and mafic granulites, in both cases doe to a retrograde evolution. The crystal chemistry and site populations of two Al-rich and one Al-poor specimen were determined by X-ray single crystal diffraction. The most relevant structural differences are the decrease in M1 and M2 polyhedral dimensions, and an increase in TB, both induced by Al. The high degree of Fe 2+ -Mg order (k D between 0.017 and 0.023) is coherent with slow cooling. Closure temperatures of the ordering process are between 214 ° and 243 ° C, with cooling rates in the range expected for a regional metamorphic terrane