Gene Expression Of Inflammatory Molecules Research Articles

Nuclear factor κB overactivation in the intervertebral disc leads to macrophage recruitment and severe disc degeneration.

Persistent inflammation has been associated with severe disc degeneration (DD). This study investigated the effect of prolonged nuclear factor κB (NF-κB) activation in DD. Using an inducible mouse model, we genetically targeted cells expressing aggrecan, a primary component of the disc extra cellular matrix, for activation of the canonical NF-κB pathway. Prolonged NF-κB activation led to severe structural degeneration accompanied by increases in gene expression of inflammatory molecules (Il1b, Cox2, Il6, and Nos2), chemokines (Mcp1 and Mif), and catabolic enzymes (Mmp3, Mmp9, and Adamts4). Increased recruitment of proinflammatory (F4/80+,CD38+) and inflammatory resolving (F4/80+,CD206+) macrophages was observed within caudal discs. We found that the secretome of inflamed caudal disc cells increased macrophage migration and inflammatory activation. Lumbar discs did not exhibit phenotypic changes, suggestive of regional spinal differences in response to inflammatory genetic overactivation. Results suggest prolonged NF-κB activation can induce severe DD through increases in inflammatory cytokines, chemotactic proteins, catabolic enzymes, and the recruitment and activation of macrophage cell populations.

Indian spices boost the immune system against COVID-19

Since 12th December 2019, the epidemic outbreak of an unknown acute respiratory tract infection has emerged in Wuhan City, China. The World Health Organization (WHO) discovered that this outbreak was caused by the 2019 novel coronavirus (2019-nCoV) or the extreme acute respiratory coronavirus 2 syndrome (SARS-CoV-2). No vaccine or no specific anti-viral treatment against COVID-19 has been made available so far. Therefore, COVID-19 can be prevented by enhancing the body's immune system and fighting off the symptoms. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is one of the essential transcription factors in humans which are responsible for the regulation of immune cell, synthesis of pro-inflammatory cytokines and gene expression of inflammatory molecules and reactive oxygen species (ROS). Phytochemicals derived from Indian spices (turmeric, garlic, ginger, etc.) can modulate the gene expression in the NF-κB pathway. In the midst of pandemic COVID-19, adding spices to food (eg. ‘Rasam’) can boost up the body's immune system. Indian spices are emerging as potential agents for the prevention of COVID-19.

Sex Specific Determinants in Osteoarthritis: A Systematic Review of Preclinical Studies.

Osteoarthritis (OA) is a highly prevalent joint disease that primarily affects about 10% of the world’s population over 60 years old. The purpose of this study is to systematically review the preclinical studies regarding sex differences in OA, with particular attention to the molecular aspect and gene expression, but also to the histopathological aspects. Three databases (PubMed, Scopus, and Web of Knowledge) were screened for eligible studies. In vitro and in vivo papers written in English, published in the last 11 years (2009–2020) were eligible. Participants were preclinical studies, including cell cultures and animal models of OA, evaluating sex differences. Independent extraction of articles and quality assessments were performed by two authors using predefined data fields and specific tools (Animals in Research Reporting In Vivo Experiments (ARRIVE) guideline and Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) tool). Twenty-three studies were included in the review: 4 in vitro studies, 18 in vivo studies, and 1 both in vitro and in vivo study. From in vitro works, sex differences were found in the gene expression of inflammatory molecules, hormonal receptors, and in responsiveness to hormonal stimulation. In vivo research showed a great heterogeneity of animal models mainly focused on the histopathological aspects rather than on the analysis of sex-related molecular mechanisms. This review highlights that many gaps in knowledge still exist; improvementsin the selection and reporting of animal models, the use of advanced in vitro models, and multiomics analyses might contribute to developing a personalized gender-based medicine.

Effect of omega‐3 and omega‐6 fatty acids on nitric oxide and IL‐6 production by RAW 264.7 macrophages

Chronic low‐grade inflammation, or meta‐inflammation, which may occur in obese subjects, is related to the infiltration of macrophages into adipose tissue (AT) as adipocyte number and size increase. The increased count in fat cells is correlated with an increase in adipose tissue macrophages (ATMs). These ATMs can secrete pro‐inflammatory cytokines which can have effects on the surrounding adipocytes. Omega‐3 fatty acids (FAs) reduce inflammation through direct competition with omega‐6 FAs, the precursors of the inflammatory eicosanoids, as well as through changes in gene expression of inflammatory molecules. Less is known about the effects of omega FAs on pro‐inflammatory cytokine production by immune cells, such as macrophages. The objective of this research was to determine the effects of omega‐3 and omega‐6 FAs on the production of nitric oxide (NO), a signaling molecule secreted by activated macrophages which is involved in mediating inflammation and production of the pro‐inflammatory cytokine interleukin‐6 (IL‐6), in lipopolysaccharide (LPS)‐activated macrophages. RAW 264.7 (murine macrophages) were incubated with 100 μM of sodium salts of linoleic acid (LA; C18:2, omega‐6), arachidonic acid (ARA; C20:4, omega‐6), or eicosapentaenoic acid (EPA; C20:5, omega‐3) for 24 h. After pre‐incubation with FAs macrophages were stimulated with 0.01 μg/ml of LPS for 6 h. Conditioned media was collected for NO and IL‐6 quantification via Griess assay and ELISA, respectively. FA treatments were cytoprotective, resulting in higher percentages of live cells than both control and LPS‐treated cells (Trypan blue exclusion method; p<0.0001, two‐way ANOVA). LPS‐stimulated macrophages pre‐incubated with LA, ARA, or EPA produced less NO than LPS‐stimulated macrophages without FA pre‐incubation (p=0.0001, two‐way ANOVA). Pre‐incubation of macrophages with LA, ARA, or EPA decreased IL‐6 production in comparison to LPS‐stimulated macrophages, with omega‐6 FAs having a more pronounced effect than omega‐3 FAs (p<0.0001, two‐way ANOVA). In conclusion, both omega‐6 and ‐3 FAs decreased activation of murine macrophages and decreased production of the inflammatory molecules NO and IL‐6, with omega‐6 FAs having a more marked effect than omega‐3 FAs. The stronger anti‐inflammatory effect of omega‐6 FAs on murine macrophages, compared to omega‐3 FAs, warrants further investigation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Anti-atherogenic and anti-inflammatory properties of glucagon-like peptide-1, glucose-dependent insulinotropic polypepide, and dipeptidyl peptidase-4 inhibitors in experimental animals.

We reported that native incretins, liraglutide and dipeptidyl peptidase‐4 inhibitors (DPP‐4i) all confer an anti‐atherosclerotic effect in apolipoprotein E‐null (Apoe −/−) mice. We confirmed the anti‐atherogenic property of incretin‐related agents in the mouse wire injury model, in which the neointimal formation in the femoral artery is remarkably suppressed. Furthermore, we showed that DPP‐4i substantially suppresses plaque formation in coronary arteries with a marked reduction in the accumulation of macrophages in cholesterol‐fed rabbits. DPP‐4i showed an anti‐atherosclerotic effect in Apoe −/− mice mainly through the actions of glucagon‐like peptide‐1 and glucose‐dependent insulinotropic polypepide. However, the dual incretin receptor antagonists partially attenuated the suppressive effect of DPP‐4i on atherosclerosis in diabetic Apoe −/− mice, suggesting an incretin‐independent mechanism. Exendin‐4 and glucose‐dependent insulinotropic polypepide elicited cyclic adenosine monophosphate generation, and suppressed the lipopolysaccharide‐induced gene expression of inflammatory molecules, such as interleukin‐1β, interleukin‐6 and tumor necrosis factor‐α, in U937 human monocytes. This suppressive effect, however, was attenuated by an inhibitor of adenylate cyclase and mimicked by 8‐bromo‐cyclic adenosine monophosphate or forskolin. DPP‐4i substantially suppressed the lipopolysaccharide‐induced expression of inflammatory cytokines without affecting cyclic adenosine monophosphate generation or cell proliferation. DPP‐4i more strongly suppressed the lipopolysaccharide‐induced gene expression of inflammatory molecules than incretins, most likely through inactivation of CD26. Glucagon‐like peptide‐1 and glucose‐dependent insulinotropic polypepide suppressed oxidized low‐density lipoprotein‐induced macrophage foam cell formation in a receptor‐dependent manner, which was associated with the downregulation of acyl‐coenzyme A cholesterol acyltransferase‐1 and CD36, as well as the up‐regulation of adenosine triphosphate‐binding cassette transporter A1. Our studies strongly suggest that incretin‐related agents have favorable effects on macrophage‐driven atherosclerosis in experimental animals.

Abstract 12161: Toll-like Receptor 9 Plays a Pivotal Role in Angiotensin II-induced Atherosclerosis

Background: Chronic vascular inflammation causes atherosclerosis. Toll-like receptor (TLR) 9 recognizes bacterial unmethylated DNA and plays a role in innate defense, whereas it can also provoke inflammation in response to fragmented DNA released from damaged mammalian cells. However, the role of TLR9 in the development of atherosclerosis remains unknown. Here, we investigated whether genetic deletion of TLR9 attenuates atherosclerosis in apolipoprotein E knockout (ApoE KO) mice. Methods and Results: We generated TLR9/ApoE double knockout (dKO) mice by crossing ApoE KO mice and TLR9 KO mice, and infused angiotensin II (Ang II) (1000 ng/kg/min) subcutaneously for 28 days using osmotic pump. There were no differences in blood pressure and plasma lipid levels between ApoE KO mice and TLR9/ApoE dKO mice. Ang II infusion significantly increased plasma levels of single-stranded DNA and double-stranded DNA, endogenous ligands of TLR9, in both strains of mice (P<0.001, respectively). TLR9/ApoE dKO mice showed less atherosclerotic lesion progression in the aortic arch and less lipid accumulation in atherosclerotic plaques in the aortic root compared with ApoE KO mice (P<0.05, respectively). TLR9/ApoE dKO mice also showed less gene expression of inflammatory molecules (e.g., ICAM-1, MCP-1) in the abdominal aorta. The result of cell sorting experiment using atherosclerotic aorta obtained from Ang II-infused ApoE KO mice demonstrated that macrophage population mainly expressed TLR9 in this tissue. We, therefore, performed in vitro experiments using thioglycolate-stimulated peritoneal macrophages. CpG ODN, agonistic oligonucleotide for TLR9, markedly promoted the expression of inflammatory molecules (e.g., ICAM-1 and MCP-1) in wild-type macrophages but not in TLR9 KO macrophages. Furthermore, conditioned medium (CM) of human umbilical vein endothelial cells (HUVECs) treated with Ang II promoted gene expression of ICAM-1 and TNF-α in THP-1 macrophages compared with CM of non-treated HUVECs. Conclusion: Our results suggested that Ang II infusion activates the TLR9 signaling in macrophages, leading to the promotion of vascular inflammation and atherosclerosis. TLR9 may serve as a potential therapeutic target for atherosclerotic diseases.

Gene Expression-Based Predictive Models of Graft Versus Host Disease-Associated Dry Eye.

To develop a predictive model based on inflammatory gene mRNA expression in conjunctival cells of graft versus host disease (GvHD)-associated dry eye (DE) patients, as well as to find meaningful correlations between gene signals and clinical signs. Twenty GvHD-DE patients and 14 healthy controls were recruited. Patients discontinued medications for 1 week before examination. Dry eye-related symptoms and signs were recorded, and conjunctival epithelial cells were collected by impression cytology after spending 20 minutes under standard conditions within a Controlled Environmental Research Laboratory. Gene expression of inflammatory molecules was determined by polymerase chain reaction, and the results were correlated with clinical signs. Shrinkage discriminant analysis, support vector machine, and k-nearest neighbor classifier methods were used to develop predictive models that were validated considering accuracy, calibration, and discriminant capability. Out of the 84 genes analyzed, 34 showed significant differences in expression. IL-6, IL-9, CCL24, CCL18, IL-10, IFN-γ, and CCL2 were highly increased (>6-fold); 26 genes were moderately upregulated (2- to 6-fold), whereas EGFR was downregulated (2.63 fold) in GvHD-DE samples. A panel based on EGFR, IL-6, IL-9, and NAMPT had an area under the receiver operating characteristic curve of 0.994, a sensitivity of 100%, and a specificity of 92.9%. EGFR expression correlated negatively with ocular surface damage markers, while IL-6, IL-9, and NAMPT correlated positively with these tests. EGFR, IL-6, IL-9, and NAMPT have the greatest potential as diagnostic biomarkers, with excellent sensitivity, specificity, and clinical relevance to the ocular surface status of GvHD.

Salt-inducible kinase 3 deficiency exacerbates lipopolysaccharide-induced endotoxin shock accompanied by increased levels of pro-inflammatory molecules in mice.

Macrophages play important roles in the innate immune system during infection and systemic inflammation. When bacterial lipopolysaccharide (LPS) binds to Toll-like receptor 4 on macrophages, several signalling cascades co-operatively up-regulate gene expression of inflammatory molecules. The present study aimed to examine whether salt-inducible kinase [SIK, a member of the AMP-activated protein kinase (AMPK) family] could contribute to the regulation of immune signal not only in cultured macrophages, but also in vivo. LPS up-regulated SIK3 expression in murine RAW264.7 macrophages and exogenously over-expressed SIK3 negatively regulated the expression of inflammatory molecules [interleukin-6 (IL-6), nitric oxide (NO) and IL-12p40] in RAW264.7 macrophages. Conversely, these inflammatory molecule levels were up-regulated in SIK3-deficient thioglycollate-elicited peritoneal macrophages (TEPM), despite no impairment of the classical signalling cascades. Forced expression of SIK3 in SIK3-deficient TEPM suppressed the levels of the above-mentioned inflammatory molecules. LPS injection (10 mg/kg) led to the death of all SIK3-knockout (KO) mice within 48 hr after treatment, whereas only one mouse died in the SIK1-KO (n = 8), SIK2-KO (n = 9) and wild-type (n = 8 or 9) groups. In addition, SIK3-KO bone marrow transplantation increased LPS sensitivity of the recipient wild-type mice, which was accompanied by an increased level of circulating IL-6. These results suggest that SIK3 is a unique negative regulator that suppresses inflammatory molecule gene expression in LPS-stimulated macrophages.

Condition-specific role of colonic inflammatory molecules in persistent functional colorectal hypersensitivity in the mouse.

A low-level inflammation has been hypothesized to mediate visceral hypersensitivity in functional bowel disorders that persist after or even in the absence of gut inflammation. We aimed to test the efficacy of a steroidal anti-inflammatory treatment, and identify local inflammatory molecules mediating post- and non-inflammatory colorectal hypersensitivity using two mouse models. Visceromotor responses to colorectal distension were quantified as a measure of colorectal sensitivity. On day 1, mice received intracolonic saline (control), trinitrobenzenesulfonic acid (postinflammatory on day 15), or acidified hypertonic saline (non-inflammatory). Colorectal sensitivity before (day 10) and after (day 15) 4-day dexamethasone (Dex) treatment was compared, and colonic gene expression of inflammatory molecules was quantified. Dexamethasone effectively inhibited gene expression of inflammatory molecules such as interleukin (IL)-1β and mast cell protease-1 in the colon, but did not attenuate colorectal hypersensitivity in either model. Gene expression of inflammatory molecules in the colon did not differ between control and the non-inflammatory model, but the postinflammatory model showed increased IL-10 and tight junction protein 2, and decreased IL-6, transforming growth factor (TGF)-β, a precursor of β-endorphin, occludin, and mucin 2. While no common molecule explained colorectal hypersensitivity in these models, hypersensitivity was positively correlated with TGF-β2 mRNA in control, and with IL-1β, inhibin βA, and prostaglandin E2 synthase in the Dex-treated postinflammatory model. In the non-inflammatory model, cyclooxygenase-2 mRNA was negatively correlated with colorectal sensitivity. These results suggest that persistent functional colorectal hypersensitivity is mediated by condition-specific mediators whose gene expression in the colon is not inevitably sensitive to steroidal anti-inflammatory treatment.

Abstract 470: Aip1 Suppresses Atherosclerosis by Limiting Hyperlipidemia-Induced Inflammation and Vascular Endothelial Dysfunction

Objective- Apoptosis signal-regulating kinase 1 (ASK1)-interacting protein-1 (AIP1) is a signaling adaptor molecule implicated in stress and apoptotic signaling induced by proinflammatory mediators. However, its function in atherosclerosis has not been established. In the present study, we use AIP1-null (AIP1-/-) mice to examine its effect on atherosclerotic lesions in an ApoE-null (ApoE-/-) mouse model of atherosclerosis. Methods and Results- ApoE-/- control mice developed atherosclerosis in the aortic roots and descending aortas upon Western-type diet for 10 weeks, while the atherosclerotic lesions are significantly augmented in ApoE-/-AIP1-/- double knockout (DKO) mice. DKO mice show increases in plasma inflammatory cytokines with no significant alterations in body weight, total cholesterol levels or lipoprotein profiles. Aortas in DKO mice show increased inflammation and endothelial cell (EC) dysfunction with NF-κB activity, correlating with increased accumulation of macrophages in the lesion area. Importantly, macrophages from DKO donors are not sufficient to augment inflammatory responses and atherogenesis when transferred to ApoE-KO recipients. Mechanistic studies suggest that AIP1 is highly expressed in aortic EC but not in macrophages, and AIP1 deletion in EC significantly enhance oxidized LDL-induced NF-κB signaling, gene expression of inflammatory molecules and monocyte adhesion, suggesting that vascular EC are responsible for the increased inflammatory responses observed in DKO mice. Conclusion- Our data demonstrate that loss of AIP1 in aortic EC primarily contributes to the exacerbated lesion expansion in the ApoE-/-AIP1-/- mice, revealing an important role of AIP1 in limiting inflammation, EC dysfunction and atherosclerosis.

AIP1 Suppresses Atherosclerosis by Limiting Hyperlipidemia-Induced Inflammation and Vascular Endothelial Dysfunction

Apoptosis signal-regulating kinase 1-interacting protein-1 (AIP1) is a signaling adaptor molecule implicated in stress and apoptotic signaling induced by proinflammatory mediators. However, its function in atherosclerosis has not been established. In the present study, we use AIP1-null (AIP1(-/-)) mice to examine its effect on atherosclerotic lesions in an apolipoprotein E-null (ApoE(-/-)) mouse model of atherosclerosis. ApoE(-/-) control mice developed atherosclerosis in the aortic roots and descending aortas on Western-type diet for 10 weeks, whereas the atherosclerotic lesions are significantly augmented in ApoE(-/-)AIP1(-/-) double knockout (DKO) mice. DKO mice show increases in plasma inflammatory cytokines with no significant alterations in body weight, total cholesterol levels, or lipoprotein profiles. Aortas in DKO mice show increased inflammation and endothelial cell (EC) dysfunction with nuclear factor-κB activity, correlating with increased accumulation of macrophages in the lesion area. Importantly, macrophages from DKO donors are not sufficient to augment inflammatory responses and atherogenesis when transferred to ApoE-KO recipients. Mechanistic studies suggest that AIP1 is highly expressed in aortic EC, but not in macrophages, and AIP1 deletion in EC significantly enhance oxidized low-density lipoprotein-induced nuclear factor-κB signaling, gene expression of inflammatory molecules, and monocyte adhesion, suggesting that vascular EC are responsible for the increased inflammatory responses observed in DKO mice. Our data demonstrate that loss of AIP1 in aortic EC primarily contributes to the exacerbated lesion expansion in the ApoE(-/-)AIP1(-/-) mice, revealing an important role of AIP1 in limiting inflammation, EC dysfunction, and atherosclerosis.

Gene Expression of Inflammatory Molecules in Circulating Lymphocytes from Arsenic-Exposed Human Subjects

Gene Expression of Inflammatory Molecules in Circulating Lymphocytes from Arsenic-Exposed Human Subjects

Gene Expression of Inflammatory Molecules in Circulating Lymphocytes from Arsenic-Exposed Human Subjects

Gene Expression of Inflammatory Molecules in Circulating Lymphocytes from Arsenic-Exposed Human Subjects