Overexpression Of Proinflammatory Cytokines Research Articles

Chronic Alcohol Ingestion and Predisposition to Lung “Cirrhosis”

Although liver is the organ most often associated with the damaging effects of chronic alcohol abuse, other organs may also be affected. In the past decade, data emerged linking chronic alcohol intake to lung dysfunction. However, the mechanisms by which alcohol affects the lung remain incompletely elucidated. In this issue, Sueblinvong and colleagues explore the effect of chronic alcohol intake in the well-known rodent model of bleomycin-induced lung injury. This represents a review of their article and a commentary on its findings in relation to current knowledge in the field. The investigators found that chronic alcohol intake increased lung fibrosis in the bleomycin-model of lung injury. This effect was related to increased production of transforming growth factor β (TGFβ) and expression of α-smooth muscle actin. Diet supplementation with S-adenosylmethionine greatly reduced the effect. These data strengthen published reports linking chronic alcohol intake with TGFβ overproduction and lung disrepair after injury, while implicating oxidant stress as a critical mediator of these effects. A review of Sueblinvong and colleagues' article and the literature strongly suggests that the lung is a target for alcohol, and that chronic alcohol intake may predispose the lung to disrepair after injury. The overexpression of pro-fibrotic growth factors and pro-inflammatory cytokines, and the generation of oxidant stress may lead to lung cellular dysfunction, aberrant tissue remodeling, and loss of lung function. These events may represent targets for intervention, but translational studies evaluating their role in humans are desperately needed.

Dietary l-arginine supplementation attenuates lipopolysaccharide-induced inflammatory response in broiler chickens

In the present study, two experiments were conducted to investigate the effect of dietary L-arginine (Arg) supplementation on the inflammatory response and innate immunity of broiler chickens. Expt 1 was designed as a 2 × 3 factorial arrangement (n 8 cages/treatment; n 6 birds/cage) with three dietary Arg concentrations (1.05, 1.42 and 1.90%) and two immune treatments (injection of lipopolysaccharide (LPS) or saline) given at an interval of 48 h between 14 and 21 d of age. In Expt 2, correlation between dietary Arg concentration (0.99, 1.39, 1.76, 2.13 or 2.53%) and percentage of circulating B cells (percentage of circulating lymphocytes) was determined. In Expt 1, LPS injection decreased body-weight gain and feed intake and increased feed conversion ratio of the challenged broilers (14-21 d; P< 0.05). LPS injection suppressed (P< 0.05) the percentages of splenic CD11+ and B cells (percentages of splenic lymphocytes) and phagocytic activity of splenic heterophils and macrophages; Arg supplementation linearly decreased the percentages of CD11+, CD14+ and B cells in the spleen (P< 0.10). LPS injection increased (P< 0.05) the expression of IL-1β and IL-6 mRNA in the spleen and caecal tonsils. Arginine supplementation decreased (P< 0.05) the expression of IL-1β, Toll-like receptor 4 (TLR4) and PPAR-γ mRNA in the spleen and IL-1β, IL-10, TLR4 and NF-κB mRNA in the caecal tonsils. In Expt 2, increasing dietary Arg concentrations linearly and quadratically reduced the percentage of circulating B cells (P< 0.01). Collectively, Arg supplementation attenuated the overexpression of pro-inflammatory cytokines probably through the suppression of the TLR4 pathway and CD14+ cell percentage. Furthermore, excessive Arg supplementation (1.76%) suppressed the percentages of circulating and splenic B cells.

L-Carnitine Attenuates the Development of Kidney Fibrosis in Hypertensive Rats by Upregulating PPAR-

The development of renal fibrosis is a consequence of arterial hypertension. L-carnitine plays an essential role in the β-oxidation of fatty acids, and we have previously demonstrated hypotensive, antioxidant, and anti-inflammatory effects of L-carnitine in arterial hypertension. This work aims to analyze the effect of L-carnitine on renal fibrosis and to explore the participation of peroxisome-proliferator activated receptor (PPAR)-γ in this effect. Four groups or rats were used: control, treated with L-carnitine, treated with L-NAME, and treated with L-carnitine + L-NAME. Cultured rat kidney cells were also used to examine the role of PPAR-γ in L-carnitine effect. An increase in the expression of collagen, transforming growth factor beta 1 (TGF-β1), connective tissue growth factor (CTGF), Nox2, and Nox4 was found in the kidney of L-NAME-treated rats. Hypertensive rats presented with an expansion of renal fibrotic areas, which was also accompanied by overexpression of proinflammatory cytokines, interleukin (IL)-1β, and IL-6. A reduction in the expression of PPAR-γ and in that of anti-inflammatory IL-10 was found in the kidney of these rats. Simultaneous treatment with L-carnitine attenuated the renal fibrosis (which correlated with a reduction of plasma TGF-β1 levels) and the pro-oxidative and proinflammatory status reported in L-NAME groups, with a concomitant increase in the expression of PPAR-γ. Furthermore, the antifibrotic effect of L-carnitine could be blocked by PPAR-γ inhibition. This study confirms the efficacy of L-carnitine against hypertension-associated renal fibrosis from in vivo and in vitro studies and suggests that the L-carnitine effect occurs in a PPAR-γ-dependent manner.

Calcitriol May Down-Regulate mRNA Over-Expression of Toll-Like Receptor-2 and -4,LL-37and Proinflammatory Cytokines in Cultured Human Keratinocytes

BackgroundAlthough vitamin D analogs have been used in the topical treatment of psoriasis, their mechanisms of action are not well understand. Calcitriol, the hormonally active vitamin D3 metabolite, has been demonstrated to exert immunomodulatory effects in the skin by down-regulating the expression of Toll-like receptors (TLRs) and proinflammatory cytokines.ObjectiveWe investigated the effects of calcitriol on the expression of TLR2, TLR4, antimicrobial peptide LL-37, and proinflammatory cytokines in cultured human keratinocytes.MethodsThe mRNA expression levels of TLR2, TLR4, tumor necrosis factor α (TNF-α), interleukin (IL)-1β and LL-37 in cultured human keratinocytes were measured by real-time polymerase chain reaction (PCR) and reverse transcription (RT). Furthermore, we measured supernatant TNF-α levels by an enzyme-linked immunosorbent assay (ELISA) to confirm the effects of calcitriol on TLR2 and TLR4.ResultsAs measured by RT-PCR and real-time PCR, calcitriol was found to suppress the lipopolysaccharide- and ultraviolet B radiation-mediated induction of expression of TLRs, LL-37 and proinflammatory cytokines such as TNF-α and IL-1β in normal human keratinocytes. The supernatant TNF-α levels measured by ELISA were also suppressed after treatment with calcitriol.ConclusionCalcitriol may down-regulate inflammatory stated over-expression of LL-37 and proinflammatory cytokines.

Mechanism of uptake of ZnO nanoparticles and inflammatory responses in macrophages require PI3K mediated MAPKs signaling

The inflammatory responses after exposure to zinc oxide nanoparticles (ZNPs) are known, however, the molecular mechanisms and direct consequences of particle uptake are still unclear. Dose and time-dependent increase in the uptake of ZNPs by macrophages has been observed by flow cytometry. Macrophages treated with ZNPs showed a significantly enhanced phagocytic activity. Inhibition of different internalization receptors caused a reduction in uptake of ZNPs in macrophages. The strongest inhibition in internalization was observed by blocking clathrin, caveolae and scavenger receptor mediated endocytic pathways. However, FcR and complement receptor-mediated phagocytic pathways also contributed significantly to control. Further, exposure of primary macrophages to ZNPs (2.5μg/ml) caused (i) significant enhancement of Ras, PI3K, (ii) enhanced phosphorylation and subsequent activation of its downstream signaling pathways via ERK1/2, p38 and JNK MAPKs (iii) overexpression of c-Jun, c-Fos and NF-κB. Our results demonstrate that ZNPs induce the generation of reactive nitrogen species and overexpression of Cox-2, iNOS, pro-inflammatory cytokines (IL-6, IFN-γ, TNF-α, IL-17 and regulatory cytokine IL-10) and MAPKs which were found to be inhibited after blocking internalization of ZNPs through caveolae receptor pathway. These results indicate that ZNPs are internalized through caveolae pathway and the inflammatory responses involve PI3K mediated MAPKs signaling cascade.

Fitting silicone hydrogel contact lenses with special diameters in a microcornea with micro and dislocated pupil

Purpose: Sarcoidosis is a chronic, multisystem granulomatous inflammatory disorder of unknown etiology. It is characterised by infiltration of tissues with macrophages and Th1/Th17lymphocytes, which culminates in the formation of noncaseating (nonnecrotizing) epithelioid cell granuloma in the affected organ. The author presents a case report of a patient who presented with bilateral granulomatous anterior uveitis as the presenting feature of ocular sarcoidosis. Method: A 40-year-old woman presented to the office with a chief complaint of pain, photophobia, tearing, redness, and cloudy vision in both eyes of 7 days duration. Clinical evaluation revealed bilateral granulomatous anterior uveitis with inflammatory ocular hypertension. Laboratory investigation demonstrated raised levels of serum Angiotensin-Converting Enzyme (ACE) and chest radiography revealed moderate bilateral pulmonary interstitial disease. The diagnosis was re-evaluated as presumed sarcoidosis-associated granulomatous anterior uveitis in line with the diagnostic criteria of the International Workshop on Ocular Sarcoidosis (IWOS). Result: A successful resolution of the granulomatous anterior uveitis with inflammatory ocular hypertension was achieved with topical therapy with corticosteroid, cycloplegia and ocular hypotensive agent. Theocular condition resolvedwithout any complications within 6 weeks. Conclusion: Sarcoid-associated anterior uveitis is an immunemediated sight-threatening eye condition. The inflammatory response in ocular sarcoidosis is characterised by an overexpression of proinflammatory cytokines and chemokines. Because Th1and Th17derived cytokines play a role in the pathomechanism of ocular sarcoidosis, early diagnosis and intervention with immunosuppressive therapy are vital to reducing ocular morbidity.

Role of nuclear factor of activated T cells 1 in the pathogenesis of osteoarthritis

Osteoarthritis (OA) is the most common form of joint disease in middle-aged individuals and the elderly. Previous studies have shown that the overexpression of matrix-degrading proteinases and proinflammatory cytokines is associated with the degradation of osteoarthritic cartilage. However, the transcription factors involved remain unclear. The present study aimed to determine the expression levels of nuclear factor of activated T cells 1 (NFAT1), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in patients with OA, and to validate the role of NFAT1 in the pathogenesis of OA. The expression levels of NFAT1, IL-1β and TNF-α in chondrocytes in the cartilage of patients with OA and healthy individuals were evaluated using western blot analysis. A luciferase reporter assay was performed to determine the activity of NFAT1 in primary human chondrocytes that were transfected with pNFAT1-luc plasmid and stimulated by IL-1β. An enzyme-linked immunosorbent assay was performed to detect the levels of TNF-α, matrix metalloproteinase (MMP)-1, MMP-3 and MMP-9 in the supernatant of cultured chondrocytes in which the NFAT1 was silenced. The expression levels of NFAT1, IL-1β and TNF-α in the cartilage of patients with OA were higher than those of the controls. IL-1β induced the expression of NFAT1 in primary chondrocytes. The expression levels of TNF-α, MMP-1, -3 and -9 promoted by IL-1β were decreased in NFAT1-silenced chondrocytes. In conclusion, NFAT1 may be important in the pathogenesis of OA and calcineurin-NFAT inhibitors may be potential effective agents for the treatment of OA.

Involvement of Endoplasmic Reticulum Stress in Albuminuria Induced Inflammasome Activation in Renal Proximal Tubular Cells

Albuminuria contributes to the progression of tubulointerstitial fibrosis. Although it has been demonstrated that ongoing albuminuria leads to tubular injury manifested by the overexpression of numerous proinflammatory cytokines, the mechanism remains largely unknown. In this study, we found that the inflammasome activation which has been recognized as one of the cornerstones of intracellular surveillance system was associated with the severity of albuminuria in the renal biopsies specimens. In vitro, bovine serum albumin (BSA) could also induce the activation of NLRP3 inflammasome in the cultured kidney epithelial cells (NRK-52E). Since there was a significant overlap of NLRP3 with the ER marker calreticulin, the ER stress provoked by BSA seemed to play a crucial role in the activation of inflammasome. Here, we demonstrated that the chemical chaperone taurine-conjugated ursodeoxycholic acid (TUDCA) which was proved to be an enhancer for the adaptive capacity of ER could attenuate the inflammasome activation induced by albuminuria not only in vitro but also in diabetic nephropathy. Taken together, these data suggested that ER stress seemed to play an important role in albuminuria-induced inflammasome activation, elimination of ER stress via TUDCA might hold promise as a novel avenue for preventing inflammasome activation ameliorating kidney epithelial cells injury induced by albuminuria.

Experimental Substantiation of the Efficiency of Antioxidant Therapy for Post-Traumatic Arthritis

Experimental post-traumatic arthritis was accompanied by overexpression of pro-inflammatory cytokines and imbalance in the collagen metabolism. Intraarticular injection of mexidol prevented free-radical degradation of collagen structures, contributed to activation of collagen synthesis and cartilage repair, and reduced the severity of inflammatory changes in the joint and the degree of secondary alteration of tissue structures.

AB0136 Induction of reg ia, a new auto-antigen in sjögren’s syndrome patients, in salivary duct epithelial cells by interleukin-6 and -11

BackgroundThe regenerating gene, Reg, was originally isolated from a rat regenerating islet cDNA library, and its human homologue was named REG Iα. Reg gene product acts as a growth factor...

CNF1 Increases Brain Energy Level, Counteracts Neuroinflammatory Markers and Rescues Cognitive Deficits in a Murine Model of Alzheimer's Disease

Overexpression of pro-inflammatory cytokines and cellular energy failure are associated with neuroinflammatory disorders, such as Alzheimer's disease. Transgenic mice homozygous for human ApoE4 gene, a well known AD and atherosclerosis animal model, show decreased levels of ATP, increased inflammatory cytokines level and accumulation of beta amyloid in the brain. All these findings are considered responsible for triggering cognitive decline. We have demonstrated that a single administration of the bacterial E. coli protein toxin CNF1 to aged apoE4 mice, beside inducing a strong amelioration of both spatial and emotional memory deficits, favored the cell energy restore through an increment of ATP content. This was accompanied by a modulation of cerebral Rho and Rac1 activity. Furthermore, CNF1 decreased the levels of beta amyloid accumulation and interleukin-1β expression in the hippocampus. Altogether, these data suggest that the pharmacological modulation of Rho GTPases by CNF1 can improve memory performances in an animal model of Alzheimer's disease via a control of neuroinflammation and a rescue of systemic energy homeostasis.

Abstract 3954: Regulation of pro-angiogenic and pro-inflammatory cytokines in tumor cells: Implication of epithelial-to-mesenchymal transitions .

Abstract Increasing data suggest a contribution of epithelial-to-mesenchymal transitions in specific steps of the metastatic cascade, particularly in tumor invasion and intravasation, and in the release of circulating tumor cells (CTCs) in the blood stream. The completion of these steps largely relies on interactions disseminating tumor cells establish with host cells. Because cytokines are particularly implicated in tumor-host cross-talks, we have examined whether EMT programs could be involved in the regulation of pro-inflammatory and pro-angiogenic cytokines which could favor angiogenesis, intravasation and CTC release. To investigate EMT implication in cytokine regulation, we used two cell lines, MDA-MB-468 (a mammary adenocarcinoma cell line) and A549 (a lung carcinoma cell line). These cell lines were shown to undergo EMT-changes following EGF and TGF-β treatment respectively, characterized by an upregulation of vimentin, snail and slug, and a downregulation of E-cadherin. A cytokine array on cell supernatant showed that EMT induction in these cell lines is associated with the upregulation of a consistant panel of cytokines. We then confirmed that interleukin-8 (IL-8), interleukin-6 (IL-6) and Granulocyte Monocyte-Colony Stimulating Factor (GM-CSF) are upregulated during EMT in the two carcinoma cell lines by RT-qPCR and ELISA. Emphasizing a functional role of such regulations, conditioned medium from EMT-induced MDA-468 cells was shown to stimulate HUVEC cell migration in a boyden chamber assay. In order to investigate whether slug and snail EMT transcription factors are involved in the regulation of IL-8, IL-6 and GM-CSF, we used both siRNA and cDNA transfection approaches. Combined repression of snail and slug inhibited the EMT-associated upregulation of IL-8, IL-6 and GM-CSF, suggesting that snail and slug are necessary for this process. Ectopic expression of each transcription factor was not sufficient to induce cytokines expression. However, ectopic expression of a proteasome degradation-resistant mutant of snail was sufficient to induce expression of IL-6, IL-8 and GM-CSF, suggesting that the stabilization of snail is necessary to regulate the expression of these cytokines. We thus here demonstrate that EMT is associated with an overexpression of specific pro-angiogenic and pro-inflammatory cytokines. We further emphasize a role of snail and slug transcription factors in this regulatory process. These data suggest that EMT programs could directly contribute to the stimulation of angiogenesis and inflammatory cell recruitment in the tumor microenvironment. The impact of such regulations on angiogenesis and the metastatic spread is currently under investigation. Citation Format: Meggy Suarez-Carmona, Morgane Bourcy, Jean-Michel Foidart, Philippe Delvenne, Agnès Noël, Myriam Polette, Christine Gilles. Regulation of pro-angiogenic and pro-inflammatory cytokines in tumor cells: Implication of epithelial-to-mesenchymal transitions . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3954. doi:10.1158/1538-7445.AM2013-3954

Heterozygosity at the A625C Polymorphic Site of the MyD88 Gene Is Associated with Mycobacterium bovis Infection in Cattle

The study demonstrates that in cattle, animals heterozygous at the MyD88 A625C polymorphic marker have a 5-fold reduced risk for active pulmonary tuberculosis (odds ratio [OR] = 0.19; P = 6 × 10(-12)). The reduced risk, however, does not extend to animals with latent pulmonary tuberculosis (OR = 0.83; P = 0.40). Heterozygosity at the A625C single nucleotide polymorphism is associated with intermediate levels of tumor necrosis factor alpha, gamma interferon, and nitric oxide synthase (NOS). Accordingly, deficiency as well as overexpression of proinflammatory cytokines or NOS favor tuberculosis, while heterozygosity provides the animals with the optimal level of inflammation.

Comparative effect of two kinds of cardioplegia on endothelial activation and inflammatory response in immature heart during cardiopulmonary bypass

Objective To compare the effect of two kinds of cardioplegia on endothelial activation and inflammatory response in immature heart during cardiopulmonary bypass (CPB).Methods Fourteen piglets,randomized to histidihe-tryptophan-ketoglutarate (HTK) group (H group) and blood cardioplegia group (B group),were put on CPB.The coronary artery and ventricular tissues after CPB were assayed for the contents of endothelial nitrogen oxygen synthetic enzyme (eNOS),vascular endothelial growth factor (VEGF),inducible nitrogen oxygen synthetic enzyme (iNOS),tumor necrotic factor-α (TNF-α),interleukin-1β (IL-1β) and IL-2.Results There were no significant differences in the contents of eNOS and VEGF in coronary vessels and iNOS,TNF-α,IL-1 β and IL-2 in myocardium between H group and B group (P ＞ O.05).Conclusion Compared with blood cardioplegia,HTK solution can preserve the contents of eNOS and VEGF in coronary endothelium and Prevent the over-expression of iNOS and pro-inflammatory cytokines in myocardium. Key words: Cardioplegia; Cardiopulmonary bypass; Immature heart; Inflammatory response

Combination therapy of methotrexate with ellagic acid attenuates the over expression of Pro-inflammatory Cytokines and modulates antioxidant status in collagen induced arthritis

Event Abstract Back to Event Combination therapy of methotrexate with ellagic acid attenuates the over expression of Pro-inflammatory Cytokines and modulates antioxidant status in collagen induced arthritis Sadiq Umar1, 2*, Sayeed Ahmad3, Chandra K. Katiyar4 and Haider A. Khan1 1 Department of toxicology, Jamia Hamdard, India 2 Jamia Millia Islamia, Department of Biotechnology, India 3 Department of Pharmacognosy, Jamia Hamdard, India 4 Ayurvedic Research Lab., Dabur Research centre, Ghaziabad, U.P, India The inclusion of antioxidant for the treatment of arthritis, especially under the therapy with immune-suppressant, is motivated because antioxidant plays an essential role in disease progression and moreover, immunosuppressive treatment suffers redox homeostasis balance of the organism. The present study was designed to evaluate the combinatory effect of methotrexate (MTX) and ellagic acid (EA) on the progression of collagen induced arthritis. Arthritis was induced in Wistar rats by collagen induced arthritis (CIA) method. CIA rats were treated with methotrexate (1 mg / kg / week) and EGCG (60 mg / kg) daily, and combination of MTX and EGCG for a period of 28 days. Paw swelling changes, scoring and histopathological was assessed to evaluate the antiarthritic effect. The effects of treatment in the rats were assessed by biochemical (articular elastase, MPO, LPO, GSH, Catalase, SOD and NO) in joint tissue homogenate to observe the modulation of antioxidant and inflammatory status, inflammatory mediators (IL-1β, TNF-α, IL-10, and PGE2) and histological studies in joints. MTX and EA combination potentiated the antiarthritic (decrease of hind paw volume and scoring) and the antioxidant effect (GSH, Catalase, SOD) as well as suppression of lipid peroxidation. Combination therapy of MTX and EA significantly inhibited the development phase of arthritis, which is supported by histopathological and attenuation of pro inflammatory cytokines. EA act as potent antioxidant and immunomodulator, suggesting that combined administration of MTX along with EA suppressed the development of arthritic progression in rats. Acknowledgements Author is grateful to ICMR, GOI for providing Research Associateship (RA). Keywords: Ellagic Acid, Rheumatoid arthritis, Inflammation, Cytokines, articular elastase Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Umar S, Ahmad S, Katiyar CK and Khan HA (2013). Combination therapy of methotrexate with ellagic acid attenuates the over expression of Pro-inflammatory Cytokines and modulates antioxidant status in collagen induced arthritis. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00795 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 14 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Sadiq Umar, Department of toxicology, Jamia Hamdard, New Delhi, Delhi, 110062, India, sadiq36@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Sadiq Umar Sayeed Ahmad Chandra K Katiyar Haider A Khan Google Sadiq Umar Sayeed Ahmad Chandra K Katiyar Haider A Khan Google Scholar Sadiq Umar Sayeed Ahmad Chandra K Katiyar Haider A Khan PubMed Sadiq Umar Sayeed Ahmad Chandra K Katiyar Haider A Khan Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Physiological regulation of pro-inflammatory cytokines expression in rat cardiovascular tissues by sympathetic nervous system and angiotensin II

Pro-inflammatory cytokines regulation by sympathetic nervous system (SNS) and angiotensin II (ANG II) was widely described in cardiovascular system, but the role of such neuro-humoral interaction needs further investigation in this context. We tested SNS-ANG II interaction on IL-6 and TNF-α mRNA expression in left ventricle and aorta from normotensive rats by sympathectomy with guanethidine and blockade of the ANG II AT1 receptors (AT1R) antagonist with losartan. mRNA synthesis of IL-6 and TNF-α were performed by Q-RT-PCR. In the left ventricle, IL-6 mRNA increased by 63% (p < 0.01) after sympathectomy, still unchanged after losartan treatment and decreased by 38% (p < 0.05) after combined treatment. TNF-α mRNA decreased by 44% (p < 0.01), only after combined treatment. In the aorta, IL-6 mRNA increased equally by 65% (p < 0.05) after sympathectomy or losartan treatment. TNF-α mRNA decreased by 28, 41, and 42% (p < 0.05) after sympathectomy, losartan and combined treatments, respectively. Our data suggest that ANG II stimulates directly (via AT1R) and indirectly (via SNS) IL-6 mRNA synthesis in left ventricle and aorta and TNF-α mRNA in left ventricle. ANG II seems unable to influence directly TNF-α mRNA synthesis in the aorta but can stimulate this cytokine via SNS. The results are relevant to prevent or reduce proinflammatory cytokines overexpression seen in cardiovascular diseases.

Silencing the expression of connexin 43 decreases inflammation and joint destruction in experimental arthritis

The objective of the present study was to determine whether the expression of connexin 43 (Cx43) effected on inflammatory conditions in rat fibroblast-like synoviocytes (FLS) and on rat model of rheumatoid arthritis (RA). The expression of Cx43 in rat FLS stimulated with lipopolysaccharide (LPS) was confirmed by real-time reverse transcriptase polymerase chain reaction (RT-PCR). The effects of small-interfering RNA targeting Cx43 (siCx43) on pro-inflammatory cytokines and chemokine were assessed by real-time RT-PCR and enzyme-linked immunosorbent assay (ELISA). The therapeutic and side effects of siCx43 in a rat model of collagen-induced arthritis (CIA) were examined by in vivo electroporation method. LPS markedly enhanced Cx43 gene expression in rat FLS, with transfection of siCx43 suppressing the over-expression of pro-inflammatory cytokines and the chemokine. Treatment of CIA rats with siCx43 significantly ameliorated paw swelling, and significantly reduced histological arthritis scores and radiographic scores. In histological appearance of rat ankle joints, siCx43 treatment significantly decreased the number of tartrate-resistant acid phosphatase (TRAP)-positive (osteoclast-like) cells. These findings indicated that siCx43 had anti-inflammatory effects in rat FLS and efficiently inhibited the development of CIA. Cx43 may play an important role in the pathophysiology of RA, and may be a potential target molecule for novel RA therapies.

Angiostatic Properties of Sulindac and Celecoxib in the Experimentally Induced Inflammatory Colorectal Cancer

Initiation of various cancers has been observed to be regulated via a prolonged inflammatory state in the tissues. However, molecular role of such a localized inflammation is not clear in the advanced stages of colorectal cancer. In this study, we have elaborated the role of various pro- and anti-inflammatory cytokines, transcription, and angiogenic factors in the progression of the 1,2-dimethylhydrazine dihydrochloride (DMH)-induced late phage colorectal cancer and also observed the chemopreventive role of the two non-steroidal anti-inflammatory drugs (NSAIDs), viz., Sulindac and Celecoxib. Carcinogenic changes were observed with morphological and histopathological studies, whereas mRNA and protein regulations of various biomolecules were identified via RT- or qRT-PCR, western blot and immunofluorescence analysis, respectively. Activity of inducible nitric oxide (NO) and cyclooxygenase-2 enzymes were analyzed using standard NO assay and prostaglandin E2 immunoassay, whereas activities of matrix metalloproteinases (MMP-2 and-9) were identified by gelatin zymography. Flowcytometry was performed for the relative quantification of the apoptotic events. Molecular docking studies of Sulindac and Celecoxib were also performed with different target proteins to observe their putative mechanisms of action. As a result, we found that DMH-treated animals were having over-expression of various pro-inflammatory cytokines (IL-1β, IL-2, and IFNγ), aberrant nuclear localization of activated cell survival transcription factors (NF-κB and Stat3) along with the increased incidence of activated angiogenic factors (MMP-2 and MMP-9) suggesting a marked role of inflammation in the tumor progression. However, NSAIDs co-administration has significantly reduced the angiogenic potential of the growing neoplasm.

The TG-interacting Factor TGIF1 Regulates Stress-induced Proinflammatory Phenotype of Endothelial Cells

The endothelium contributes to the control of the tissue inflammatory response following stress and in particular after exposure to ionizing radiation. We previously showed that the TG-interacting factor 1 (TGIF1) plays a role in radiation-induced normal tissue injury. In this study we hypothesized that this protein could play a role in inflammation. The role of TGIF1 in the stress-induced proinflammatory phenotype was investigated in human endothelial cells. In HUVECs ionizing radiation induces TGIF1 expression as well as a proinflammatory phenotype associated with up-regulation of IL-6, IL-8, CXCL1, MIP-2, and MCP-1. TGIF1 overexpression enhances the radiation-induced proinflammatory phenotype whereas TGIF1 silencing limits both the TNF-α- and radiation-induced overexpression of proinflammatory cytokines. Interestingly, in vivo, in radiation-induced intestinal inflammation in mice, TGIF1 genetic deficiency is associated with a reduced radiation-induced overexpression of proinflammatory molecules. In HUVECs, TNF-α- and radiation-induced NF-κB pathway activation is not influenced by TGIF1 expression, whereas TGIF1 knockdown inhibits both TNF-α- and radiation-induced p38 MAPK pathway activation. This study demonstrates that TGIF1 plays a role in TNF-α- and radiation-induced inflammation and suggests that it could be a target in limiting this event in the vascular compartment.

Expression profiling of Rab GTPases reveals the involvement of Rab20 and Rab32 in acute brain inflammation in mice

Rab GTPases have emerged as central regulators of vesicle trafficking and are essential for cytokine production during the pathogenesis of neuroinflammation. To characterize the roles of different Rab proteins in brain inflammation, we used quantitative PCR (qPCR) to examine the expression profiles of all members of the Rab family in an experimental model of brain inflammation in mice. We found that Rab20 and Rab32 were substantially up-regulated during the acute phase of inflammation. The increased expression of Rab20 was also confirmed by immunostaining of inflamed brains at different timepoints. The concomitant overexpression of Rabs (Rab20 and Rab32) and early response proinflammatory cytokines (TNF-α and IL-1β) suggested that these Rabs may be important for subsequent inflammatory responses in brain. Furthermore, we found that the expression of certain Rabs was dramatically reduced in cultured primary microglia, which was not observed in the in vivo profiling. In N9, a microglial cell line, however, there was no increase in the expression of Rab20 or Rab32, but Rab3c was significantly overexpressed. These results collectively indicate that Rabs may participate in inflammatory response in microglia during brain inflammation. The differential regulation of individual Rabs in different experimental systems is a caveat for the analysis of Rab functions.