Inhibited Monocyte Chemoattractant Protein-1 Expression Research Articles

Sirt1 suppresses MCP-1 production during the intervertebral disc degeneration by inactivating AP-1 subunits c-Fos/c-Jun.

The anti-inflammatory effect of Sirtuin 1 (Sirt1) during intervertebral disc degeneration (IDD) has been widely confirmed. Monocyte chemoattractant protein-1 (MCP-1) activation is the initiating inflammatory response associated with the IDD. However, whether Sirt1 suppresses MCP-1 in the intervertebral disc is unclear. The MCP-1 and Sirt1 protein expression in the degenerated and non-degenerated NP tissues were compared by immunohistochemistry (IHC). We induced nucleus pulposus (NP) cell degeneration by IL-1β and mediated cellular Sirt1 expression through the Sirt1 activator resveratrol (Res) or inhibitor Nicotinamide (Nico). In addition, the inhibitors of MCP-1 and Activator protein 1 (AP-1) were also used in cell culture. The function of NP cells was determined by the type II collagen and Cell Counting Kit-8 (CCK-8) assay. We assessed the Sirt1 and MCP-1 expression by the Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR). The AP-1 activity was valued by the phosphorylation of its components c-Fos, and c-Jun. Both in vivo and in vitro experimental results indicated that MCP-1 was upregulated in the degenerated condition, which was opposite to Sirt1 expression. Res suppressed AP-1, the phosphorylation of c-Fos/c-Jun, and the MCP-1 expression. On the contrary, Sirt1 downregulation by Nico aggravated the phosphorylation of c-Fos/c-Jun and MCP-1 expression. However, the MCP-1 suppression did not affect the Sirt1 and AP-1 levels. The destruction of AP-1 activation also inhibited MCP-1 expression but not Sirt1. The upregulation of Sirt1 and suppression of MCP-1 improved the type II collagen expression and cell viability, which was injured by IL-1β. Sirt1 suppresses the MCP-1 production in the degenerated NP cells by suppressing the phosphorylation of the AP-1 subunits c-Fos and c-Jun.

Curcumin Inhibits Monocyte Chemoattractant Protein-1 Expression in TNF-α induced Astrocytes Through AMPK Pathway.

Curcumin, a phenolic pigment, plays an inhibitory role in astrocytes activation, a key step for neuropathic pain (NP). The present study aimed to investigate the mechanism behind the therapeutic effect of Curcumin on NP in vitro. Specifically, we investigated the inhibitory effect of Curcumin on tumor necrosis factor-α (TNF-α)-induced astrocyte migration. We also studied the effects of Curcumin on monocyte chemoattractant protein-1(MCP-1) expression and activity, as well as super oxide dismutase-2 (SOD2) expression and activity in TNF-α-induced astrocytes. Additionally, we investigated whether the adenosine-monophosphate-activated protein kinase signaling (AMPK) pathway was involved in this process. Our data demonstrated that Curcumin inhibited TNF-α-induced astrocytes migration, decreased MCP-1 expression, and up-regulated SOD2 expression in TNF-α-induced astrocytes in vitro. Our study also indicated that this process was mediated through the AMPK signaling pathway, as addition of Curcumin significantly increased the level of phosphorylated AMPK protein. Furthermore, the specific AMPK activator AICAR (5-aminoimidazole-4-carboxamide 1-D-ribofuranoside) mimicked the effects of Curcumin, whereas a selective AMPK inhibitor Compound C (also called dorsomorphin) partially blocked its function. These results could shed light on understanding of the molecular basis for the inhibition of Curcumin on MCP-1 expression during the process of astrocyte activation, and provide a molecular mechanism for using Curcumin in neuropathic pain.

Naringenin suppresses macrophage infiltration into adipose tissue in an early phase of high-fat diet-induced obesity

Obese adipose tissue is characterized by increased macrophage infiltration, which results in chronic inflammation in adipose tissue and leads to obesity-related diseases such as type 2 diabetes mellitus and atherosclerosis. The regulation of macrophage infiltration into adipose tissue is an important strategy for preventing and treating obesity-related diseases. In this study, we report that naringenin, a citrus flavonoid, suppressed macrophage infiltration into adipose tissue induced by short-term (14days) feeding of a high-fat diet in mice; although naringenin did not show any differences in high-fat diet-induced changes of serum biochemical parameters in this short administration period. Naringenin suppressed monocyte chemoattractant protein-1 (MCP-1) in adipose tissue, and this effect was mediated in part through inhibition of c-Jun NH2-terminal kinase pathway. Naringenin also inhibited MCP-1 expression in adipocytes, macrophages, and a co-culture of adipocytes and macrophages. Our results suggest a mechanism by which daily consumption of naringenin may exhibit preventive effects on obesity-related diseases.

Curcumin inhibits monocyte chemoattractant protein-1 expression and enhances cholesterol efflux by suppressing the c-Jun N-terminal kinase pathway in macrophage.

To investigate the effect of curcumin on monocyte chemoattractant protein 1 (MCP-1) production and reverse cholesterol transport (RCT) in macrophage induced by oxidation low-density lipoprotein (ox-LDL), and to identify the signal pathways involved. The macrophages were treated with ox-LDL and various concentrations of curcumin simultaneously. The MCP-1 expression was measured by enzyme-linked immunosorbent assay. The apoAI-mediated cholesterol efflux was measured by (3)H-cholesterol-labeled counting radioactivity. The activation of intracellular signaling pathways was studied by Western blotting. Curcumin decreased the production of MCP-1 induced by ox-LDL in macrophages. MCP-1 expression was restrained by the inhibition of c-Jun N-terminal kinase (JNK) pathway (SP600125) and NF-κB pathway (BAY11-7082). Curcumin suppressed the phosphorylation of JNK and activation of NF-κB. Curcumin also enhanced RCT via up-regulating the expression of liver X receptor alpha (LXRα), ATP-binding cassette transporter A1 (ABCA1) and scavenger receptor class B type I (SR-BI). Additionally, the inhibition of JNK (SP600125) increased cholesterol efflux and increased the expression of ABCA1 and SR-BI, but had no effect on LXRα. Curcumin suppresses MCP-1 production induced by ox-LDL via the JNK pathway and NK-κB pathway, while enhances cholesterol efflux in macrophage via suppressing the JNK pathway and activating the LXR-ABCA1/SR-BI pathway, which indicate that the vascular protective effect of curcumin is related to anti-inflammation and anti-atherosclerosis.

Aldosterone mediates glomerular inflammation in experimental mesangial proliferative glomerulonephritis

The renoprotection of the mineralocorticoid receptor antagonist (MRA) is considered to be mainly via its antifibrotic activity, and the possibility that it may also have antiinflammatory effects has not been studied. We tested the hypothesis that MRA might influence the inflammatory changes that accompany experimental glomerular injury. Administration of vehicle (control) or a selective MRA, eplerenone (50 mg/kg x 2 times/day) was started 7 days (-7d) before induction of anti-Thy-1.1 glomerulonephritis. Kidney samples were evaluated serially over a 12-day period for the presence of cell proliferation, macrophage infiltration, mesangial cell phenotypic activation and expression of the chemokine monocyte chemoattractant protein-1 (MCP-1). MRA did not prevent the mesangiolysis associated with anti-Thy-1 antibody. However, MRA significantly inhibited MCP-1 expression, glomerular macrophage infiltration and mesangial phenotypic activation (alpha-smooth muscle actin expression). MRA alters glomerular inflammation and mesangial cell activation in experimental glomerular injury. MRA may be a novel way to treat acute glomerular diseases.

968 REGULATION OF MCP-1 EXPRESSION THROUGH ANGIOTENSIN II TYPE 1 RECEPTOR IN CASTRATION-RESISTANT PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Basic Research V1 Apr 2012968 REGULATION OF MCP-1 EXPRESSION THROUGH ANGIOTENSIN II TYPE 1 RECEPTOR IN CASTRATION-RESISTANT PROSTATE CANCER Suguru Shirotake, Akira Miyajima, Takeo Kosaka, Nobuyuki Tanaka, Eiji Kikuchi, and Mototsugu Oya Suguru ShirotakeSuguru Shirotake Tokyo, Japan More articles by this author , Akira MiyajimaAkira Miyajima Tokyo, Japan More articles by this author , Takeo KosakaTakeo Kosaka Tokyo, Japan More articles by this author , Nobuyuki TanakaNobuyuki Tanaka Tokyo, Japan More articles by this author , Eiji KikuchiEiji Kikuchi Tokyo, Japan More articles by this author , and Mototsugu OyaMototsugu Oya Tokyo, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1067AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Tumor associated macrophage infiltration and the main regulator, monocyte chemoattractant protein-1 (MCP-1) expression, have been reported to be associated with a poor prognosis in prostate cancer (CaP). However, the mechanism of regulation of MCP-1 in CaP is still unclear. Since angiotensin II (AngII) type 1 receptor (AT1R) is known to be one of the regulators of MCP-1 in hypertensive disease and we have already investigated the correlation between AT1R and tumor angiogenesis in CaP, here we examined the regulation of MCP-1 through AT1R in CaP. METHODS Specimens were obtained from 138 CaP patients; androgen dependent CaP in 121 cases obtained by prostatectomy and castration resistant CaP (CRPC) in 17 cases obtained by autopsy. All specimens were immunostained for MCP-1 and CD68+ macrophages (Mφ). Clinicopathological characteristics were reviewed retrospectively. We also investigated the regulation of MCP-1 expression through AT1R using 3 human CaP cell lines; LNCaP, C4-2, and C4-2AT6. CV11974 and TCV116 are active metabolites of candesartan and were used for the in vitro and in vivo experiments. RESULTS Specimens with a high Gleason score (7≤) and advanced stage (pT3≤), and those of CRPC had significantly higher MCP-1 expression and higher Mφ infiltration than low malignant CaP. Kaplan-Meier curves showed that the 5-year PSA recurrence-free survival rate was 77.0% in the low MCP-1 group vs 54.5% in the high MCP-1 group (p=0.017). Multivariate analysis revealed that preoperative high PSA (15≤) (risk ratio, 2.48) and high MCP-1 (risk ratio, 2.20) were independent risk factors for PSA recurrence. Higher expression of MCP-1 and AT1R in C4-2 and C4-2AT6 cells than in LNCaP cells was observed. AngII induced significantly higher MCP-1 in C4-2AT6 cells than in LNCaP cells, while MCP-1 expression was reduced by CV11974. C4-2AT6 tumors treated with TCV116 also exhibited significantly lower MCP-1 expression and Mφ infiltration than the control tumors. In C4-2AT6 cells, immunoblotting demonstrated that CV11974 inhibited the phosphorylation of Akt by AngII stimulation, but that the MAPK and JAK/STAT pathways were not activated by AngII. MCP-1 induced by AngII was suppressed by PI3K inhibitor in C4-2AT6 cells. CONCLUSIONS Both high MCP-1 and high Mφ in CaP specimens correlated with a high PSA recurrence rate. AT1R blockade inhibited MCP-1 expression through the PI3K/Akt pathway and also suppressed Mφ infiltration in CRPC. AT1R is one of the regulators of MCP-1 expression, and may potentially become a new therapeutic target molecule for the treatment of CRPC. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e394 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Suguru Shirotake Tokyo, Japan More articles by this author Akira Miyajima Tokyo, Japan More articles by this author Takeo Kosaka Tokyo, Japan More articles by this author Nobuyuki Tanaka Tokyo, Japan More articles by this author Eiji Kikuchi Tokyo, Japan More articles by this author Mototsugu Oya Tokyo, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Knockdown of VEGF receptor-1 (VEGFR-1) impairs macrophage infiltration, angiogenesis and growth of clear cell renal cell carcinoma (CRCC)

Angiogenesis is essential for tumor growth and metastasis. VEGF has been shown to be a central player in this process. The biological activity of VEGF is mainly mediated by two tyrosine kinase receptors, VEGFR-1 and VEGFR-2. While increasing evidence suggests that VEGF/VEGFR-1 signaling is crucial for tumor angiogenesis, its molecular mechanism is not well understood. Here we show that VEGFR-1 knockdown dramatically inhibits tumor growth. This inhibition is associated with significant decrease of tumor VEGF levels and tumor angiogenesis as well as an increased tumor necrosis. Moreover, we demonstrate that VEGF in CRCC tumors is mainly produced by tumor stromal cells instead of the tumor cells themselves. It has been shown that macrophages constitute a significant part of tumor stromal cells and produce a large amount of VEGF. We therefore examined the macrophage infiltration in the xenograft tumors. Remarkably, VEGFR-1 knockdown attenuates the tumor macrophages infiltration. To understand the mechanism, we investigated the impact of VEGFR-1 knockdown on the expression of monocyte chemoattractant protein-1 (MCP-1), one of the main chemoattractants for macrophages. Significantly, VEGFR-1 knockdown inhibits MCP-1 expression of CRCC cells. Taken together, these data indicate that VEGF/VEGFR-1 signaling plays an essential role in initiating tumor angiogenesis by regulating MCP-1 expression, which in turn, attracts macrophages infiltration and VEGF production. Thus, these studies suggest that blockade of VEGFR-1 function may provide a tumor-specific, VEGF-based therapeutic strategy for treatment of CRCC.

TRB3: an oxidant stress-induced pseudokinase with a potential to negatively modulate MCP-1 cytokine in diabetic nephropathy

TRB3: an oxidant stress-induced pseudokinase with a potential to negatively modulate MCP-1 cytokine in diabetic nephropathy

A Novel Role for the Glucocorticoid Receptor in the Regulation of Monocyte Chemoattractant Protein-1 mRNA Stability

Monocyte chemoattractant protein-1 (MCP-1) plays an important role in attracting monocytes to sites of inflammation and is the dominant mediator of macrophage accumulation in atherosclerotic plaques. We have previously shown that glucocorticoids inhibit the secretion of MCP-1 in arterial smooth muscle cells (SMC) by markedly decreasing MCP-1 mRNA stability. We now report that the destabilization of MCP-1 mRNA is mediated by the glucocorticoid receptor (GR). The GR antagonist, RU486, blocked the effect of the glucocorticoid dexamethasone (Dex) on MCP-1 mRNA stability in SMC culture. Using a previously reported in vitro mRNA gel shift and stability assay, antibodies to the GR blocked the ability of cytoplasmic extracts from Dex-treated SMC to decay MCP-1 mRNA. Recombinant human GR (rhGR) bound in a concentration-dependent manner to in vitro transcribed MCP-1 mRNA, whereas other members of the steroid hormone receptor family did not. Binding of GR to MCP-1 mRNA was specific as it was not found to bind other mRNAs. Immunoprecipitation of GR in extracts from Dex-treated SMC followed by real-time reverse transcription-PCR demonstrated that endogenous GR was bound specifically to MCP-1 mRNA. The addition of exogenous rhGR blocked the ability of extracts from Dex-treated SMC to degrade MCP-1 mRNA, suggesting that exogenous rhGR can compete with an endogenous GR-containing degradative complex. These data suggest a novel role for the GR in binding to and facilitating mRNA degradation. These results provide novel insights into GR function and may provide a new approach to attenuate the inflammatory response mediated by MCP-1.

LY294002 inhibits monocyte chemoattractant protein-1 expression through a phosphatidylinositol 3-kinase-independent mechanism

The effects of LY294002 (LY29) and wortmannin (WM), inhibitors of phosphatidylinositol 3-kinase (PI3K), on monocyte chemoattractant protein-1 (MCP-1) expression by human umbilical vein endothelial cells were investigated. Complete inhibition of interleukin (IL)-1β-induced Akt phosphorylation occurred at 50 μM LY29 or 100 nM WM. At these concentrations, LY29, but not WM, significantly inhibited constitutive and IL-1β-induced MCP-1 expression at both protein and mRNA levels. LY303511 (LY30), an inactive analogue of LY29, also inhibited MCP-1 expression. LY29 and LY30 inhibited activation of nuclear factor-κB (NF-κB). These results suggest that LY29 inhibits MCP-1 expression at least in part via suppression of NF-κB, independent of PI3K, and the structure of LY29 and LY30 may be a novel template for development of new anti-inflammatory drugs.

Exogenous Nitric Oxide Inhibits Tumor Necrosis Factor-Alpha- or Interleukin-1-Beta-Induced Monocyte Chemoattractant Protein-1 Expression in Human Mesangial Cells

Monocyte chemoattractant protein-1 (MCP-1) plays an important role in glomerulonephritis and nitric oxide (NO) exerts a variety of renal pathophysiological effects. We investigated the effect of exogenous NO on pro-inflammatory cytokine-induced MCP-1 expression in human mesangial cells and its signal transduction pathway. Cells were pretreated with NO donors such as 3-morpholino-sydnonimine (SIN-1) or nitroprusside, and then stimulated with tumor necrosis factor-α (TNF-α) or interleukin-1β (IL-1β). MCP-1 expression of mRNA and protein were measured by Northern blot analysis and ELISA. NF-ĸB binding activity was determined by electrophoretic mobility shift assay. Degradation of IĸB-α protein was assessed by Western blot analysis. SIN-1 inhibited TNF-α- or IL-1β-induced MCP-1 mRNA expression in a dose-dependent manner and also suppressed the MCP-1 protein expression. Nitroprusside inhibited the MCP-1 mRNA expression as well. SIN-1 dose dependently inhibited the TNF-α- or IL-1β-induced NF-ĸB binding activity and suppressed the TNF-α-induced degradation of IĸB-α. Analogue of cGMP (8-bromo-cGMP) had no significant effect on TNF-α-induced MCP-1 mRNA expression and guanylate cyclase inhibitor (ODQ) also had no significant influence on the inhibitory effect of SIN-1. These results suggest that exogenous NO inhibits MCP-1 expression via suppression of NF-ĸB by reducing the degradation of IĸB-α and through a cGMP-independent pathway.

Negative regulation of monocyte chemoattractant protein-1 gene expression by a mouse estrogen-enhanced transcript.

A novel gene containing two typical estrogen responsive elements (ERE) was cloned from MTEC1 cells, a mouse thymus epithelial cell line that produces constitutively many chemokines. This gene is a homologue of the rat estrogen-enhanced transcript (EET) gene, and is called the mEET gene. mEET protein is expressed in cytoplasm. Addition of 17 beta-estradiol (E2) to the MTEC1 cell cultures enhanced mEET mRNA expression and, meanwhile, significantly inhibited monocyte chemoattractant protein-1 (MCP-1) production. To analyze the functional links between the expression of mEET and of MCP-1, we transfected MTEC1 cells with ERE-deleted antisense- or sense-mEET complementary (c)DNA construct and activated the transfected mEET cDNA in stable MTEC1 transfectants with doxycycline (Dox). Dox-induced activation of the mEET gene profoundly inhibited MCP-1 expression at both mRNA and protein levels and alleviated its chemotactic activity. Conversely, inactivation of the mEET gene substantially augmented MCP-1 expression. Activation of the mEET gene markedly attenuated activity of nuclear NF-kappaB. In summary, we have first demonstrated that estrogen-imposed inhibition of MCP-1 expression occurs through the activation of the mEET gene, its product suppresses nuclear NF-kappaB and negatively regulates MCP-1 gene activation.