Chemokine Monocyte Chemoattractant Protein-1 Research Articles

Ethanol-activated microglial exosomes induce MCP1 signaling mediated death of stress-regulatory proopiomelanocortin neurons in the developing hypothalamus

BackgroundMicroglia, a type of resident immune cells within the central nervous system, have been implicated in ethanol-activated neuronal death of the stress regulatory proopiomelanocortin (POMC) neuron-producing β-endorphin peptides in the hypothalamus in a postnatal rat model of fetal alcohol spectrum disorders. We determined if microglial extracellular vesicles (exosomes) are involved in the ethanol-induced neuronal death of the β-endorphin neuron via secreting elevated levels of the chemokine monocyte chemoattractant protein 1 (MCP1), a key regulator of neuroinflammation.MethodsWe employed an in vitro model, consisting of primary culture of hypothalamic microglia prepared from postnatal day 2 (PND2) rat hypothalami and treated with or without 50 mM ethanol for 24 h, and an in vivo animal model in which microglia were obtained from hypothalami of PND6 rats fed daily with 2.5 mg/kg ethanol or control milk formula for five days prior to use. Exosomes were extracted and characterized with nanosight tracking analysis (NTA), transmission electron microscopy and western blot. Chemokine multiplex immunoassay and ELISA were used for quantitative estimation of MCP1 level. Neurotoxic ability of exosome was tested using primary cultures of β-endorphin neurons and employing nucleosome assay and immunocytochemistry. Elevated plus maze, open field and restraint tests were used to assess anxiety-related behaviors.ResultsEthanol elevated MCP1 levels in microglial exosomes both in vitro and in vivo models. Ethanol-activated microglial exosomes when introduced into primary cultures of β-endorphin neurons, increased cellular levels of MCP1 and the chemokine receptor CCR2 related signaling molecules including inflammatory cytokines and apoptotic genes as well as apoptotic death of β-endorphin neurons. These effects of microglial exosomes on β-endorphin neurons were suppressed by a CCR2 antagonist RS504393. Furthermore, RS504393 when injected in postnatal rats prior to feeding with ethanol it reduced alcohol-induced β-endorphin neuronal death in the hypothalamus. RS504393 also suppressed corticosterone response to stress and anxiety-like behaviors in postnatally alcohol-fed rats during adult period.ConclusionThese data suggest that alcohol exposures during the developmental period elevates MCP1 levels in microglial exosomes that promote MCP1/CCR2 signaling to increase the apoptosis of β-endorphin neurons and resulting in hormonal and behavioral stress responses.

NIK Is a Mediator of Inflammation and Intimal Hyperplasia in Endothelial Denudation-Induced Vascular Injury

Neointimal hyperplasia is the main cause of vascular graft failure in the medium term. NFκB is a key mediator of inflammation that is activated during neointimal hyperplasia following endothelial injury. However, the molecular mechanisms involved in NFκB activation are poorly understood. NFκB may be activated through canonical (transient) and non-canonical (persistent) pathways. NFκB-inducing kinase (NIK, MAP3K14) is the upstream kinase of the non-canonical pathway. We have now explored the impact of NIK deficiency on neointimal hyperplasia following guidewire-induced endothelial cell injury and on local inflammation by comparing NIK activity–deficient alymphoplasia mice (NIKaly/aly) with control wild-type (NIK+/+) mice. Guidewire-induced endothelial cell injury caused neointimal hyperplasia and luminal stenosis and upregulated the local expression of NIK and the NFκB target chemokines monocyte chemoattractant protein-1 (MCP-1/CCL2) and chemokine ligand 5 (RANTES/CCL5). Immunohistochemistry disclosed the infiltration of the media and intima by F4/80 positive macrophages. The intima/media ratio and percentage of stenosis were milder in the NIKaly/aly than in the NIK+/+ mice. Additionally, the gene expression for MCP-1 and RANTES was lower and F4/80+ cell infiltration was milder in the NIKaly/aly than in the NIK+/+ mice. Finally, circulating MCP-1 levels were lower in the NIKaly/aly than in the NIK+/+ mice, reflecting milder systemic inflammation. In conclusion, NIK is a driver of vascular wall inflammation and stenosis following guidewire-induced endothelial cell injury. NIK targeting may be a novel therapeutic approach to limit arterial stenosis following endothelial cell injury.

DaiTongXiao improves gout nephropathy by inhibiting inflammatory response through the TLR4/MyD88/NF-κB pathway.

Introduction: Gouty nephropathy (GN) arises from factors like excessive purine intake, metabolic disorders or abnormal synthesis, and uric acid hypersaturation in the blood, leading to urate crystal deposition in kidney tissue. DaiTongXiao (DTX) is a remedy used by the Dai people of China. It shows efficacy in lowering uric acid levels and exhibits anti-inflammatory and kidney-protective properties. Methods: A GN rat model was induced using adenine and potassium oxonate. Following DTX administration, various parameters were assessed in urine, serum, and kidney tissue. Western blot analysis evaluated TLR4/MyD88/NF-κB signaling proteins, while immunofluorescence examined NF-κB nuclear expression. Results: DTX treatment improved kidney morphology, increased body weight, and kidney index and enhanced urinary levels of blood urea nitrogen (Bun), 24-h urinary protein, uric acid (UA), and allantoin in GN rats, reducing UA, Bun, creatinine (Cre), cystatin C (CysC), serum amyloid A (SAA), α1-microglobulin (MG), and β2-MG in serum analysis. Renal tissue assessments showed decreased xanthine oxidase (XOD), hydroxyproline (Hyp), α-smooth muscle actin (α-SMA), and collage type Ⅳ (COL-Ⅳ). Kidney damage severity was notably reduced. DTX lowered serum inflammatory factors like interleukin (IL) -18, tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), transforming growth factor-β1 (TGF-β1), and IL-1β in the rat serum, reducing chemokine monocyte chemoattractant protein-1 (MCP-1) and adhesion factor vascular cell adhesion molecule-1(VCAM-1). Western blotting demonstrated the downregulation of TLR4/MyD88/NF-κB pathway proteins, and immunofluorescence revealed reduced NF-κB expression in renal tissue. Discussion: DTX exhibits significant anti-GN effects by modulating TLR4/MyD88/ NF-κB pathway protein expression, reducing inflammatory factor release, and inhibiting GN progression.

Proinflammatory Activation of Monocytes in Patients with Immunoinflammatory Rheumatic Diseases.

. The objective of this study was to evaluate the proinflammatory activation of circulating monocytes in patients with IRDs. . The study involved 149 participants (53 patients with rheumatoid arthritis (RA), 45patients with systemic lupus erythematosus (SLE), 34 patients with systemic scleroderma (SSc), and 17participants without IRDs) 30 to 65 years old. Basal and lipopolysaccharide (LPS)-stimulated secretion of monocytes was studied in a primary culture of monocytes obtained from blood by immunomagnetic separation. Quantitative assessment of the cytokines tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), as well as the chemokine monocyte chemoattractant protein-1 (MCP-1) was carried out in the culture fluid by ELISA. Proinflammatory activation of monocytes was calculated as the ratio of LPS-stimulated and basal secretions. . It was shown that the basal secretion of all studied cytokines was significantly increased in all groups of patients with IRDs, except for the secretion of IL-1β in the SLE group, compared to the control. LPS-stimulated secretion of TNF-α was increased and MCP-1 was decreased in patients with IRDs compared to the control group; LPS-stimulated IL-1β secretion only in the SSc group significantly differed from the control group. In the RA group, monocyte activation was reduced for all cytokines compared to the control; in the SLE group, for TNF-α and MCP-1; in the SSc group, for MCP-1. . The decrease in proinflammatory activation of monocytes in patients with IRDs is due to a high level of basal secretion of cytokines, which can lead to disruption of the adequate immune response in these diseases and is an important link in the pathogenesis of chronic inflammation.

Neurosteroid [3α,5α]3-hydroxypregnan-20-one inhibition of chemokine monocyte chemoattractant protein-1 in alcohol-preferring rat brain neurons, microglia, and astroglia.

Neuroimmune dysfunction in alcohol use disorder (AUD) is associated with activation of myeloid differentiation primary response 88 (MyD88)-dependent Toll-like receptors (TLR) resulting in overexpression of the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2). MCP-1 overexpression in the brain is linked to anxiety, higher alcohol intake, neuronal death, and activation of microglia observed in AUD. The neurosteroid [3α,5α][3-hydroxypregnan-20-one (3α,5α-THP) has been reported as an inhibitor of MyD88-dependent TLR activation and MCP-1 overexpression in mouse and human macrophages and the brain of alcohol-preferring (P) rats. We investigated how 3α,5α-THP regulates MCP-1 expression at the cellular level in P rat nucleus accumbens (NAc) and central amygdala (CeA). We focused on neurons, microglia, and astrocytes, examining the individual voxel density of MCP-1, neuronal marker NeuN, microglial marker IBA1, astrocytic marker GFAP, and their shared voxel density, defined as intersection. Ethanol-naïve male and female P rats were perfused 1 h after IP injections of 15 mg/kg of 3α,5α-THP, or vehicle. The NAc and CeA were imaged using confocal microscopy following double-immunofluorescence staining for MCP-1 with NeuN, IBA1, and GFAP, respectively. MCP-1 intersected with NeuN predominantly and IBA1/GFAP negligibly. 3α,5α-THP reduced MCP-1 expression in NeuN-labeled cells by 38.27 ± 28.09% in male and 56.11 ± 21.46% in female NAc, also 37.99 ± 19.53% in male and 54.96 ± 30.58% in female CeA. In females, 3α,5α-THP reduced the MCP-1 within IBA1 and GFAP-labeled voxels in the NAc and CeA. Conversely, in males, 3α,5α-THP did not significantly alter the MCP-1 within IBA1 in NAc or with GFAP in the CeA. Furthermore, 3α,5α-THP decreased levels of IBA1 in both regions and sexes with no impact on GFAP or NeuN levels. Secondary analysis performed on data normalized to % control values indicated that no significant sex differences were present. These data suggest that 3α,5α-THP inhibits neuronal MCP-1 expression and decreases the proliferation of microglia in P rats. These results increase our understanding of potential mechanisms for 3α,5α-THP modulation of ethanol consumption.

The protective effects of alamandine on angiotensin II-induced cardiac dysfunction and remodeling in rats

Alamandine (Ala), a relatively recent addition to the renin-angiotensin system (RAS), is a heptapeptide sharing structural similarities with angiotensin-(1-7). This peptide has garnered significant attention due to its distinctive antihypertensive, vasodilatory, and antifibrotic effects. In this study, we sought to determine whether Ala acts as a counter-regulatory agent against angiotensin II-induced cardiac hypertrophy, fibrosis, and inflammation in rats. Male SD rats (n=6 in each group) received a two-week subcutaneous infusion of vehicle (saline), angiotensin II (ANG II, 150 ng/kg/min), or a combination of ANG II and Ala (50 ng/kg/min) via mini osmotic pumps. Left ventricular (LV) function was evaluated through echocardiography. Histological analysis of cardiac hypertrophy and fibrosis was conducted using Hematoxylin Eosin and Trichome-Mason staining, respectively. Compared to the vehicle group, treatment with Ala significantly (*p<0.05) attenuated ANG II-induced cardiac mass (659.27 ± 54.26* vs. 824.48 ± 40.38 mg) with reduced LV thickness and end-diastolic volume. Ala treatment also improved cardiac function, as indicated by reduced global longitudinal strain and myocardial performance index. Histological images revealed that Ala treatment markedly reduced cardiomyocyte area (383.63* ± 7.20 vs 490.59 ± 13.13 μm²) and fibrosis area (5.22* ± 0.93 vs. 9.64 ± 0.36%) in rats treated with ANG II, along with reduced mRNA expression of pro-hypertrophic markers such as brain natriuretic peptide (4.29 ± 0.50* vs 7.53 ± 0.68, fold change) and β-myosin heavy chain (6.11 ± 0.61* vs 10.29 ± 1.02), as well as pro-fibrosis markers collagen 1 (5.72 ± 0.59* vs 10.56 ± 1.69), fibronectin (2.18 ± 0.34* vs 4.69 ± 0.70), and α-smooth muscle actin (2.14 ± 0.42* vs 3.85 ± 0.06), when compared to the rats treated with ANG II alone. Flow cytometry analysis also showed that Ala treatment significantly reduced ANG II-induced immune responses in the heart by reducing immune cell populations including CD4, CD8 T cells, B cells and NK cells, and increasing presence of anti-inflammatory M2 monocytes. Additionally, the mRNA levels of interlukin-6, CD 68, and chemokine monocyte chemoattractant protein-1 were significantly decreased in Ala-treated rats. Furthermore, treatment with Ala reversed ANG II-induced alterations in the mRNA expression of angiotensin-converting enzyme (ACE, 2.05 ± 0.16* vs. 3.08 ± 0.25), angiotensin type 1 receptor (1.45 ± 0.22* vs. 2.56 ± 0.31), and ACE 2 (1.08 ± 0.10** vs. 0.38 ± 0.06) in the LV of the heart. Taken together, these findings demonstrate that alamandine exerts a protective effect against ANG II-induced cardiac dysfunction, hypertrophy, fibrosis, and inflammation, potentially acting as a protective component of the RAS to counteract the adverse effects of ANG II. Alamandine might be a potential therapeutic agent in the treatment of cardiovascular disorders. Supported by NIH grants R01 HL139521 & 155091. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Paricalcitol Has a Potent Anti-Inflammatory Effect in Rat Endothelial Denudation-Induced Intimal Hyperplasia.

Neointimal hyperplasia is the main cause of vascular graft failure in the medium term. Vitamin D receptor activation modulates the biology of vascular smooth muscle cells and has been reported to protect from neointimal hyperplasia following endothelial injury. However, the molecular mechanisms are poorly understood. We have now explored the impact of the selective vitamin D receptor activator, paricalcitol, on neointimal hyperplasia, following guidewire-induced endothelial cell injury in rats, and we have assessed the impact of paricalcitol or vehicle on the expression of key cell stress factors. Guidewire-induced endothelial cell injury caused neointimal hyperplasia and luminal stenosis and upregulated the expression of the growth factor growth/differentiation factor-15 (GDF-15), the cytokine receptor CD74, NFκB-inducing kinase (NIK, an upstream regulator of the proinflammatory transcription factor NFκB) and the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2). Immunohistochemistry confirmed the increased expression of the cellular proteins CD74 and NIK. Paricalcitol (administered in doses of 750 ng/kg of body weight, every other day) had a non-significant impact on neointimal hyperplasia and luminal stenosis. However, it significantly decreased GDF-15, CD74, NIK and MCP-1/CCL2 mRNA expression, which in paricalcitol-injured arteries remained within the levels found in control vehicle sham arteries. In conclusion, paricalcitol had a dramatic effect, suppressing the stress response to guidewire-induced endothelial cell injury, despite a limited impact on neointimal hyperplasia and luminal stenosis. This observation identifies novel molecular targets of paricalcitol in the vascular system, whose differential expression cannot be justified as a consequence of improved tissue injury.

Plasma and peritoneal fluid cytokine profiles in patient with Essure® implant: Towards a molecular signature?

Objective(s)Many patients with Essure® implant may experience adverse events related to the device. Although local inflammation does not appear to be the pathophysiological mechanism underlying the symptoms, systemic inflammation could play a role. In the present study, as cytokines are involved in the inflammatory process, we proposed to investigate the profile of circulating and peritoneal cytokines. Study designIn this retrospective study, we evaluated the levels of cytokines in peritoneal fluid (PF) as well as in plasma sample from three different groups: Essure® group, endometriosis group (known to be associated with immune dysregulation), and control group. ResultsThere were 60 symptomatic patients with Essure® device, 30 patients with endometriosis and a control group of 30 patients. The PF levels of Interleukin-10 (IL-10), Interleukin-6 (IL-6), and Monocyte chemoattractant protein-1 (MCP-1) were statistically higher in endometriosis group than in Essure® group and control group. The plasma level of MCP-1 was higher in Essure® group than in endometriosis group and control group. The plasma level of TNF-α was higher in Essure® group than in control group. ConclusionsThe chemokine MCP-1 as well as the pro-inflammatory TNF-α, are known to be increased in patients with fibromyalgia and chronic fatigue syndrome. Since patients with Essure® may exhibit symptoms similar to fibromyalgia, MCP-1 and TNF-α may be relevant markers in symptomatic patients with Essure®. Because of the lack of longitudinal data (no evaluation of postoperative cytokine profile and no assessment of the level of clinical improvement), other studies are needed to confirm these preliminary results.

Assessment of Monocyte Chemoattractant Protein -1 and Fertility Hormones in Iraqi Women with Polycystic Ovarian Syndrome

Polycystic ovarian syndrome (PCOS) is a well-known endocrinopathy and one of the most frequent endocrine-reproductive-metabolic syndromes in women, which can result in reduced fertility. While the actual cause is unknown, PCOS is regarded as a complicated genetic characteristic with a great degree of variability. Moreover, hormones and immune cells, including both innate and acquired immune cells, are thought to interact in PCOS. Chronic low-grade inflammation raises the risk of autoimmune disease. The study's purpose is to investigate the chemokine monocyte chemoattractant protein-1 (MCP-1) and fertility hormones in samples of women patients with polycystic ovary syndrome (PCOS) in the City of Medicine. Sixty PCOS women comprise 30 healthy control women; their average age was 20–40 years, and their weight ranged from 60 to 100 kg. The results showed an increase in the level of MCP1 in PCOS patients, but this increase was not significant (P<0.05), which was not influenced by BMI or fertility hormones. As well as elevated fertility hormones, this study, when compared to controls as well as patients with PCOS, showed a significant increase in the level of testosterone (14.63 ±2.30 nmol/L) while in control women (0.627 ±0.04), LH hormone in patients and control group (6.54 ±0.51 mIU/mL), and 2.93 ±0.18, respectively. Prolactin hormone was increased in PCOS patients (16.27 ±1.25 ng/mL) when compared to the control group \ (12.85 ±0.62). There was no significant difference in FSH hormone in women with PCOS (5.27 ±0.28 mIU/mL) compared with the control group (5.59 ±0.18).

Epstein-Barr Virus Promotes Inflammatory Cytokine Production in Human Gingival Fibroblasts

BackgroundPeriodontitis is one of the most common chronic oral inflammatory diseases. Over the past decade, herpes viruses, particularly Epstein-Barr virus (EBV), have been considered promising pathogenic candidates for periodontitis. However, the specific mechanism by which EBV contributes to the development of periodontitis is still unknown. This study aimed to explore the mechanism of EBV underlying the inflammatory response in human gingival fibroblasts (HGFs). Materials and methodsHGFs were stimulated with different concentrations of EBV (104, 105, 106, 107, and 108 DNA copies/mL) for 0, 8, 24, or 48 hours. The mRNA levels of interleukin (IL)-1β, tumour necrosis factor-α (TNF-α), IL-8, monocyte chemoattractant protein-1 (MCP-1), and Toll-like receptor 9 (TLR9) were measured using quantitative real-time polymerase chain reaction (PCR). Enzyme-linked immunosorbent assays (ELISAs) were performed for determining the mRNA and protein levels of IL-1β, TNF-α, IL-8, and MCP-1. Real-time PCR and ELISA were performed to determine the protein levels of IL-1β, TNF-α, IL-8, and MCP-1. Activation of the TLR9/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa B (NF-κB) pathway was evaluated using western blotting. ResultsThe expressions of IL-1β, TNF-α, IL-8, and MCP-1 were significantly upregulated in HGFs under EBV stimulation in a concentration- and time-dependent manner. EBV promoted TLR9 and MyD88 expression and induced NF-κB transcription. On the contrary, the upregulation of these factors and the activation of NF-κB pathway were drastically inhibited by TLR9 antagonists. ConclusionsOur findings demonstrate that EBV promotes the production of inflammatory cytokines IL-1β and TNF-α and chemokines IL-8 and MCP-1 in HGFs through the TLR9/MyD88/NF-κB pathway.

Monocyte chemoattractant protein-1 and cyclooxygenase enzymes in rats treated with drugs of abuse

The CC chemokine monocyte chemoattractant protein (MCP)-1/CCL2 and cyclo-oxygenase-2 enzyme are potent mediators of neuroinflammation. We studied the effect of treatment with Cannabis sativaresin extract and/or tramadol, two common drugs of abuse on the level of MCP-1 in brain and serum of rats. In addition, serum levels of cyclo-oxygenase 1 (COX-1) and cyclo-oxygenase 2 (COX-2) enzymes in rats given cannabis werealso determined. Cannabis sativaresin at 5, 10 or 20 mg/kg (expressed as delta-9-tetrahydrocannabinol), tramadol at 5, 10 or 20 mg/kg or tramadol at 10 mg/kg combined with cannabis at 5-20 mg/kg were given once a day, subcutaneously, for six weeks. Resultsshowed that, as compared to corresponding controls, Cannabis sativa resin produced a significant increase in MCP-1 increased inbrain tissue (29.8-56.4%) and serum (14.8-31.0%). MCP-1 showed also significant increments in brain (34.6%) and serum (22.1%) by 20 mg/kg tramadol. After the concomitant treatment with both cannabis and tramadol, MCP-1 increased in brainby 88.5-145.7%and in serum by 32.6-64.8%. Moreover, cannabis given at 20 mg/kg significantly increasedserum COX-1 and COX-2 by 362% and 58.8% compared to corresponding control values. The histopathological study of the striatum of cannabis-treated animals revealed the presence of apoptotic cells and pyknotic nuclei and congestion of blood vessels. The striatum from animals given tramadol showed neuronal perinuclear cytoplasmic vacuolations and apoptotic cells. Moreover, the combined administration of both cannabis and tramadol resulted in neuronal necrosis, vacuolation, apoptosis and pyknosis. Collectively, these results indicate that treatment with cannabis resin and/or tramadol is associated with a proinflammatory response that involves MCP-1 and cyclooxygenases and which could result in the development of brain injury.

The Clinical Relevance of Selected Cytokines in Newly Diagnosed Multiple Myeloma Patients.

Multiple myeloma (MM) is the second most common hematological neoplasm. Cytokines, chemokines, and their receptors, induced by the microenvironment of MM, participate in tumor growth, the attraction of leukocytes, cell homing, and bone destruction. This study aimed to assess the correlation between the pretreatment serum concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), angiogenic chemokine monocyte chemoattractant protein-1 (MCP-1), and vascular endothelial growth factor (VEGF) and the clinical outcomes and survival of patients newly diagnosed with MM. The study group consisted of 82 individuals. The IL-8 concentration was significantly positively correlated with the age of onset (p = 0.007), the International Staging System (ISS) stage (p = 0.03), the Eastern Cooperative Oncology Group (ECOG) performance status (p < 0.001), the degree of anemia before treatment (p < 0.0001), the degree of kidney disease (p < 0.001), and VEGF (p = 0.0364). Chemotherapy responders had significantly lower concentrations of IL-8 (p < 0.001), IL-6 (p < 0.001), and VEGF (p = 0.04) compared with non-responders. Patients with treatment-induced polyneuropathy had significantly higher levels of IL-8 (p = 0.033). Patients with a high level of IL-6 had a 2-fold higher risk of progression-free survival (PFS) reduction (17 vs. 35 months; HR = 1.89; p = 0.0078), and a more than 2.5-fold higher risk of overall survival (OS) reduction (28 vs. 78 months; HR = 2.62; p < 0.001). High levels of IL-6, IL-8, and VEGF demonstrated significant predictive values for some clinical conditions or outcomes of newly diagnosed MM patients. Patients with an early response to chemotherapy had a significantly lower concentration of these cytokines. A high pretreatment IL-6 concentration was an independent negative prognostic marker for newly diagnosed MM patients.

SAT381 Effect of Obesity on Androgen-induced Splenic Dysregulation in Polycystic Ovary Syndrome

Abstract Disclosure: S. Rezq: None. J. Basnet: None. A.M. Huffman: None. L.L. Yanes Cardozo: None. D.G. Romero: None. Background: Polycystic ovary syndrome (PCOS) is characterized by ovarian dysfunction and hyperandrogenism. Women with PCOS exhibit low-grade systemic inflammation, obesity, and immune dysregulation. The spleen plays an important role in immune function and physiological inflammation regulation. Androgens and obesity negatively impact splenic structure and function. However, the effect of androgens on spleen architecture and function and its modulation by obesogenic diet is unknown in PCOS. Using a well-established PCOS model of hyperandrogenemia we aim to test the hypothesis that androgens cause splenic dysregulations that are exacerbated by obesity. Methods: Three-week-old female mice were treated with the androgen dihydrotestosterone (DHT, 8 mg, s.c) or vehicle. Animals were fed a high-fat diet (HFD; 60% kcal fat) or a low-fat diet (LFD; 10% kcal fat) for 90 days (n=6-8/group). Body composition (EchoMRI) and spleen weight were assessed. Splenic mRNA or protein expression of multiple immune cells/regulatory markers such as chemokines ligands Cxcl12 and Lymphotoxin B (Ltb), and their corresponding receptors C-X-C chemokine receptor type 4 (CXCR4) and Ltb receptor (LTBR), as well as the chemokine monocyte chemoattractant protein-1 (MCP-1), proliferation markers Csf1, Csf2, and Csf3, the adhesion molecule Vcam1, and extracellular matrix (ECM) remodeling marker Coch were assessed by RT-qPCR or Western blot. Additionally, splenic androgen receptor (AR) protein expression was measured by Western blot, while the inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) were detected by ELISA. Results: DHT significantly (p&amp;lt;0.05) increased fat mass (6.2 ± 0.5 vs. 3.4 ± 0.3 g) and spleen weight (11.9 ± 1.6 vs. 4.9 ± 0.22 g) in LFD-fed mice, which was exacerbated by HFD. DHT-mediated splenomegaly was associated with a significant (p&amp;lt;0.05) decrease in the expression of AR (90%), the chemokine Cxcl12 (70-80%), the proliferation markers Csf1 and Csf2 (65-66%), and the ECM marker Coch (70%) in mice that were fed on both diets. PCOS mice had significantly (p&amp;lt;0.05) higher CXCR4 (4.4-4.7-fold) and LTBR (5.7-7.6-fold) expression, independent of diet. Interestingly, only the HFD-fed PCOS mice had significantly (p&amp;lt;0.05) lower splenic Vcam1 (76%), IL-6 (25%), and TNF-α (4.6 ± 0.2 vs. 5.6 ± 0.2 pg/μg protein) levels, while they also had higher MCP-1 protein levels (2-fold, p&amp;lt;0.05). Conclusion and significance: Our data show that androgens cause splenic structural and molecular alterations that are modulated by obesity. These findings could explain PCOS-associated immune dysregulation that is more predominant in obese women with PCOS. The study highlights the spleen as a novel key target organ of androgen-mediated pathology in PCOS. Furthermore, our study suggests that weight loss could potentially attenuate androgen-associated splenic derangements in PCOS. Presentation Date: Saturday, June 17, 2023

Depression and HIV: a scoping review in search of neuroimmune biomarkers.

People with HIV are at increased risk for depression, though the neurobiological mechanisms underlying this are unclear. In the last decade, there has been a substantial rise in interest in the contribution of (neuro)inflammation to depression, coupled with rapid advancements in the resolution and sensitivity of biomarker assays such as Luminex, single molecular array and newly developed positron emission tomography radioligands. Numerous pre-clinical and clinical studies have recently leveraged these next-generation immunoassays to identify biomarkers that may be associated with HIV and depression (separately), though few studies have explored these biomarkers in co-occurring HIV and depression. Using a systematic search, we detected 33 publications involving a cumulative N = 10 590 participants which tested for associations between depressive symptoms and 55 biomarkers of inflammation and related processes in participants living with HIV. Formal meta-analyses were not possible as statistical reporting in the field was highly variable; future studies must fully report test statistics and effect size estimates. The majority of included studies were carried out in the United States, with samples that were primarily older and primarily men. Substantial further work is necessary to diversify the geographical, age, and sex distribution of samples in the field. This review finds that alterations in concentrations of certain biomarkers of neuroinflammation (interleukin-6, tumour necrosis factor-α, neopterin) may influence the association between HIV and depression. Equally, the chemokines monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) or the metabolic index kynurenine:tryptophan (Kyn:Trp), which have been the focus of several studies, do not appear to be associated with depressive symptoms amongst people living with HIV, as all (MCP-1) or most (IL-8 and Kyn:Trp) available studies of these biomarkers reported non-significant associations. We propose a biomarker-driven hypothesis of the neuroimmunometabolic mechanisms that may precipitate the increased risk of depression among people with HIV. Chronically activated microglia, which trigger key neuroinflammatory cascades shown to be upregulated in people with HIV, may be the central link connecting HIV infection in the central nervous system with depressive symptoms. Findings from this review may inform research design in future studies of HIV-associated depression and enable concerted efforts towards biomarker discovery.

Serotonin transporter-deficient mice display enhanced adipose tissue inflammation after chronic high-fat diet feeding.

Serotonin is involved in leukocyte recruitment during inflammation. Deficiency of the serotonin transporter (SERT) is associated with metabolic changes in humans and mice. A possible link and interaction between the inflammatory effects of serotonin and metabolic derangements in SERT-deficient mice has not been investigated so far. SERT-deficient (Sert -/-) and wild type (WT) mice were fed a high-fat diet, starting at 8 weeks of age. Metabolic phenotyping (metabolic caging, glucose and insulin tolerance testing, body and organ weight measurements, qPCR, histology) and assessment of adipose tissue inflammation (flow cytometry, histology, qPCR) were carried out at the end of the 19-week high-fat diet feeding period. In parallel, Sert -/- and WT mice received a control diet and were analyzed either at the time point equivalent to high-fat diet feeding or as early as 8-11 weeks of age for baseline characterization. After 19 weeks of high-fat diet, Sert -/- and WT mice displayed similar whole-body and fat pad weights despite increased relative weight gain due to lower starting body weight in Sert -/-. In obese Sert -/- animals insulin resistance and liver steatosis were enhanced as compared to WT animals. Leukocyte accumulation and mRNA expression of cytokine signaling mediators were increased in epididymal adipose tissue of obese Sert -/- mice. These effects were associated with higher adipose tissue mRNA expression of the chemokine monocyte chemoattractant protein 1 and presence of monocytosis in blood with an increased proportion of pro-inflammatory Ly6C+ monocytes. By contrast, Sert -/- mice fed a control diet did not display adipose tissue inflammation. Our observations suggest that SERT deficiency in mice is associated with inflammatory processes that manifest as increased adipose tissue inflammation upon chronic high-fat diet feeding due to enhanced leukocyte recruitment.

Time- and glucose-dependent differentiation of 3T3-L1 adipocytes mimics dysfunctional adiposity

The 3T3-L1 murine adipocyte cell line remains one of the most widely used models to study the mechanisms of obesity and related pathologies. Most studies investigate such mechanisms using mature adipocytes that have been chemically induced to differentiate for 7 days in media containing 25 mM glucose. However, the dysfunctional characteristics commonly observed in obesity including adipocyte hypertrophy, increased expression of inflammatory markers, enhanced production of reactive oxygen species (ROS), increased steroidogenic enzyme expression/activity and production of steroid hormones, are not necessarily mimicked in these cells. The aim of this study was to provide an inexpensive model which represents the well-known characteristics of obesity by manipulating the time of adipocyte differentiation and increasing the concentration of glucose in the cell media. Our results showed a glucose- and time-dependent increase in adipocyte hypertrophy, ROS production and gene expression of the pro-inflammatory cytokine interleukin-6 (IL-6), as well as a time-dependent increase in lipolysis and in the gene expression of the chemokine monocyte chemoattractant protein 1 (MCP1). We also showed that gene expression of the steroidogenic enzymes 11-beta-hydroxysteroid dehydrogenase type 1 (11βHSD1), 17βHSD type 7 and 12, as well as CYP19A1 (aromatase), were significantly higher in the hypertrophic model relative to the control adipocytes differentiated using the conventional method. The increase in 11βHSD1 and 17βHSD12 expression was consistent with the enhanced conversion of cortisone and androstenedione to cortisol and testosterone, respectively. As these characteristics reflect those commonly observed in obesity, hypertrophic 3T3-L1 adipocytes are an appropriate in vitro model to study mechanisms of adipocyte dysfunction in an era where the rise in obesity incidence is a global health concern, and where access to adipose tissue from obese patients are limited.

ОЦЕНКА ЗНАЧИМОСТИ ВЗАИМОСВЯЗИ МЕЖДУ ПОКАЗАТЕЛЯМИ РЕМОДЕЛИРОВАНИЯ СОСУДОВ, ВОСПАЛИТЕЛЬНОГО ОТВЕТА И ПОЛИМОРФИЗМА ГЕНОВ ДИСФУНКЦИИ ЭНДОТЕЛИЯ У ПАЦИЕНТОВ МУЖСКОГО ПОЛА С ИНФАРКТОМ МИОКАРДА В ЗАВИСИМОСТИ ОТ НАЛИЧИЯ ИЛИ ОТСУТСТВИЯ ГИПЕРТОНИЧЕСКОГО АНАМНЕЗА

Introduction. Endothelial dysfunction, arterial hypertension and surrogate events of myocardial infarction are interrelated phenomena, while the genetic predisposition in these patients is an independent risk factor for the development of cardiovascular pathology. Aim. The aim of the study was to evaluate the significance of the interaction of associations of polymorphic loci of adhesion molecule genes (immunoglobulin VCAM1), MCP-1 chemokine, vascular endothelial growth factor (VEGFA), nitric oxide control enzyme (DDAH1), markers of the inflammatory response, vascular remodeling and endothelial dysfunction in patients with male myocardial infarction, depending on the presence or absence of arterial hypertension in history in the development of myocardial infarction. Material and methods. 667 male patients and 621 practically healthy individuals in the comparison group were examined. Patients underwent determination of general clinical parameters, serum concentrations of monocyte-chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGFA), cytokines (IL-1,6; TNF-α), molecular genetic study, measurement of media intima thickness of the common carotid artery. Selected in the group of patients with myocardial infarction, individuals with or without a history of arterial hypertension. Results and discussion. As a result of a comparative analysis of groups of patients with and without a history of arterial hypertension with a group of healthy individuals, significant differences in the frequencies of genotypes and alleles for the following polymorphic gene loci were revealed: MCP-1, VEGFA, DDAH1, VCAM1. The serum concentration of cytokines (IL-1,6; TNF-α) in patients, regardless of the presence or absence of a history of hypertension, significantly exceeded the level in the comparison group. The VEGFA index in patients with myocardial infarction with arterial hypertension was significantly increased compared to the serum level in patients with myocardial infarction without arterial hypertension. Conclusion. A higher significance of the interaction between indicators of vascular remodeling, inflammation, endothelial dysfunction gene polymorphism in the development of myocardial infarction in patients with a history of hypertension, in contrast to non-hypertensive patients with myocardial infarction, was revealed, which indicates a powerful contribution of arterial hypertension to the etiology and implementation of myocardial infarction.

Acertannin Prevented Dextran Sulfate Sodium-induced Colitis by Inhibiting the Colonic Expression of IL-23 and TNF-α in C57BL/6J Mice.

The present study investigates the effects of acertannin on colitis induced by dextran sulfate sodium (DSS) and changes in the colonic levels of the cytokines interleukin (IL)-1β, IL-6, IL-10, IL-23, tumor necrosis factor (TNF)-α, the chemokine monocyte chemoattractant protein (MCP)-1, and vascular endothelial growth factor (VEGF).We examine the following: inflammatory colitis was induced in mice by 2% DSS drinking water given ad libitum for 7 days. Red blood cell, platelets, and leukocyte counts and hematocrit (Ht), hemoglobin (Hb), and colonic cytokine and chemokine levels were measured. The disease activity index (DAI) was lower in DSS-treated mice orally administered acertannin (30 and 100 mg/kg) than in DSS-treated mice. Acertannin (100 mg/kg) inhibited reductions in the red blood cell count and Hb and Ht levels in DSS-treated mice. Acertannin prevented DDS-induced mucosal membrane ulceration of the colon and significantly inhibited the increased colonic levels of IL-23 and TNF-α. Our findings suggest that acertannin has potential as a treatment for inflammatory bowel disease (IBD).

Baicalein alleviates fibrosis and inflammation in systemic sclerosis by regulating B-cell abnormalities

BackgroundSystemic sclerosis (SSc; also known as “scleroderma”) is an autoimmune disorder characterized by extensive fibrosis, vascular changes, and immunologic dysregulation. Baicalein (phenolic flavonoid derived from Scutellaria baicalensis Georgi) has been used to treat the pathological processes of various fibrotic and inflammatory diseases. In this study, we investigated the effect of baicalein on the major pathologic characteristics of SSc: fibrosis, B-cell abnormalities, and inflammation.MethodsThe effect of baicalein on collagen accumulation and expression of fibrogenic markers in human dermal fibroblasts were analyzed. SSc mice were produced by injecting bleomycin and treated with baicalein (25, 50, or 100 mg/kg). The antifibrotic features of baicalein and its mechanisms were investigated by histologic examination, hydroxyproline assay, enzyme-linked immunosorbent assay, western blotting and flow cytometry.ResultsBaicalein (5–120 μM) significantly inhibited the accumulation of the extracellular matrix and fibroblast activation in transforming growth factor (TGF)-β1- and platelet derived growth factor (PDGF)-induced human dermal fibroblasts, as evidenced by abrogated deposition of total collagen, decreased secretion of soluble collagen, reduced collagen contraction capability and downregulation of various fibrogenesis molecules. In a bleomycin-induced model of dermal fibrosis in mice, baicalein (25–100 mg/kg) restored dermal architecture, ameliorated inflammatory infiltrates, and attenuated dermal thickness and collagen accumulation in a dose-dependent manner. According to flow cytometry, baicalein reduced the proportion of B cells (B220+ lymphocytes) and increased the proportion of memory B cells (B220+CD27+ lymphocytes) in the spleens of bleomycin-induced mice. Baicalein treatment potently attenuated serum levels of cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-17A, tumor necrosis factor-α), chemokines (monocyte chemoattractant protein-1, macrophage inflammatory protein-1 beta) and autoantibodies (anti-scleroderma 70 (Scl-70), anti-polymyositis-scleroderma (PM-Scl), anti-centromeres, anti-double stranded DNA (dsDNA). In addition, baicalein treatment can significantly inhibit the activation of TGF-β1 signaling in dermal fibroblasts and bleomycin-induce mice of SSc, evidenced by reducing the expression of TGF-β1 and IL-11, as well as inhibiting both small mother against decapentaplegic homolog 3 (SMAD3) and extracellular signal-related kinase (ERK) activation.ConclusionsThese findings suggest that baicalein has therapeutic potential against SSc, exerting modulating B-cell abnormalities, anti-inflammatory effects, and antifibrosis.

Effect of astragaloside Ⅳ on angiotensin Ⅱ-induced inflammatory response of vascular endothelial cells and mechanism

This study was designed to determine the inhibitory effect of astragaloside Ⅳ(AS-Ⅳ), a principal bioactive component extracted from the Chinese medicinal Astragali Radix, on the inflammatory response of vascular endothelial cells induced by angiotensin Ⅱ(Ang Ⅱ), the most major pathogenic factor for cardiovascular diseases, and to clarify the role of calcium(Ca~(2+))/phosphatidylinosi-tol-3-kinase(PI3K)/protein kinase B(Akt)/endothelial nitric oxide synthase(eNOS)/nitric oxide(NO) pathway in the process. To be specific, human umbilical vein endothelial cells(HUVECs) were cultured in the presence of AS-Ⅳ with or without the specific inhibitor of NO synthase(NG-monomethyl-L-arginine, L-NMMA), inhibitor of PI3K/Akt signaling pathway(LY294002), or Ca~(2+)-chelating agent(ethylene glycol tetraacetic acid, EGTA) prior to Ang Ⅱ stimulation. The inhibitory effect of AS-Ⅳ on Ang Ⅱ-induced inflammatory response and the involved mechanism was determined with enzyme-linked immunosorbent assay(ELISA), cell-based ELISA assay, Western blot, and monocyte adhesion assay which determined the fluorescently labeled human monocytic cell line(THP-1) adhered to Ang Ⅱ-stimulated endothelial cells. AS-Ⅳ increased the production of NO by HUVECs in a dose-and time-dependent manner(P&lt;0.05) and raised the level of phosphorylated eNOS(P&lt;0.05). The above AS-Ⅳ-induced changes were abolished by pretreatment with L-NMMA, LY294002, or EGTA. Compared with the control group, Ang Ⅱ obviously enhanced the production and release of cytokines(tumor necrosis factor-α, interleukin-6), chemokines(monocyte chemoattractant protein-1) and adhesion molecules(intercellular adhesion molecule-1, vascular cellular adhesion molecule-1), and the number of monocytes adhered to HUVECs(P&lt;0.05), which were accompanied by the enhanced levels of phosphorylated inhibitor of nuclear factor-κBα protein and activities of nuclear factor-κB(NF-κB)(P&lt;0.05). This study also demonstrated that Ang Ⅱ-induced inflammatory response was inhibited by pretreatment with AS-Ⅳ(P&lt;0.05). In addition, the inhibitory effect of AS-Ⅳ was abrogated by pretreatment with L-NMMA, LY294002, or EGTA(P&lt;0.05). This study provides a direct link between AS-Ⅳ and Ca~(2+)/PI3K/Akt/eNOS/NO pathway in AS-Ⅳ-mediated anti-inflammatory actions in endothelial cells exposed to Ang Ⅱ. The results indicate that AS-Ⅳ attenuates endothelial cell-mediated inflammatory response induced by Ang Ⅱ via the activation of Ca~(2+)/PI3K/Akt/eNOS/NO signaling pathway.