Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P43026 (lipopolysaccharide)
 62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have investigated the effects of lipopolysaccharide (LPS) and probucol (a lipid soluble antioxidant) on the gene expression of interleukin 1 alpha and beta (IL-1 alpha and IL-1 beta), and platelet-derived growth factor A chain and B chain (PDGF-A and PDGF-B) in the human monocytic cell line U-937. Steady-state mRNA levels were measured quantitatively by the reverse transcription-polymerase chain reaction (RT-PCR) using a non-radioactive label. Cells were incubated with LPS, in the presence or absence of probucol for 20 h. The cells were harvested and RNA was then prepared, reverse-transcribed in the presence of an internal standard and subsequently amplified and labelled with digoxigenin-11-dUTP by the PCR reaction. The PCR products were subjected to agarose gel electrophoresis, blotted onto nylon membranes and visualised by immunological detection of digoxigenin followed by a chemiluminescent reaction. We found that: (1) LPS treatment of U-937 cells caused an up-regulation of gene expression of IL-1 beta and PDGF-A chain by approximately 250% and 100% respectively, although it did not stimulate the expression of IL-1 alpha nor PDGF-B chain mRNA. (2) Probucol treatment in vitro had no effect on the basal or LPS-stimulated mRNA levels of IL-1 alpha, IL-1 beta, PDGF-A and PDGF-B despite its reported activity in vivo. (3) PDGF-A and PDGF-B were expressed at a similar level in unstimulated U-937 cells approximately 10-50 copies/ng total RNA, whereas the expression of IL-1 beta mRNA was approximately 2-4 times higher than IL-1 alpha mRNA. (4) Finally, in U-937 monocytic cells the expression of IL-1 alpha and IL-1 beta, and PDGF-A and PDGF-B appear to be independently regulated.
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PMID:The effects of LPS and probucol on interleukin 1 (IL-1) and platelet-derived growth factor (PDGF) gene expression in the human monocytic cell line U-937. 831 73

A specific radioimmunoassay was employed to demonstrate that human articular cartilage and chondrocyte monolayers in organ and cell culture, respectively, produce macrophage colony-stimulating factor (M-CSF) in response to stimulation with interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF alpha) and TNF beta. Optimum doses were 10-100 U/ml for IL-1 (0.06-0.6 nM IL-1 alpha; 0.02-0.2 nM IL-1 beta) and 1-10 nM for TNF alpha. Low levels of M-CSF were observed in the supernatants of nonstimulated cultures while increased levels of M-CSF in response to IL-1 alpha and TNF alpha were detected following 2 h exposure to the cytokines. IL-1 alpha and TNF alpha did not show synergy for the production of M-CSF when both cytokines were added to cultures. Actinomycin D and cycloheximide inhibited both the basal and IL-1 alpha-induced production of M-CSF, suggesting a requirement for de novo RNA and protein synthesis. Cytokine-induced M-CSF production was also inhibited by the antiinflammatory corticosteroid, dexamethasone, but not by the cyclooxygenase inhibitor, indomethacin. The cytokines IL-4, IL-6, platelet-derived growth factor, leukemia inhibitory factor, transforming growth factor-beta and interferons -alpha and -gamma, each had little or no effect on M-CSF levels, while basic fibroblast growth factor, lipopolysaccharide, and retinoic acid were each weak stimuli. We propose that chondrocyte M-CSF production in response to IL-1 and TNF alpha, and the concurrent destruction of cartilage by these cytokines, could provide a mechanism for the chronic nature of rheumatoid disease.
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PMID:Production of macrophage colony-stimulating factor (M-CSF) by human articular cartilage and chondrocytes. Modulation by interleukin-1 and tumor necrosis factor alpha. 834 86

The retinal pigment epithelial (RPE) cell is a potent regulatory cell within the retina. It helps to maintain normal retinal activity, and following gamma interferon (IFN-gamma) exposure, it may express major histocompatibility complex class II molecules and function as an antigen-presenting cell. Since interleukin-1 (IL-1) and IL-6 are potent cytokines observed in ocular inflammatory processes, we initiated studies to evaluate conditions which enable RPE cells to produce these cytokines. Cultures of human RPE cells from two eye donors were established and characterized, and enzyme immunoassays were employed to screen for IL-1 and IL-6 production. Treatment of RPE cells with lipopolysaccharide (LPS) or recombinant tumor necrosis factor alpha, IL-1, or IFN-gamma resulted in a significant level of secretion of IL-6. In contrast, treatment with recombinant epidermal growth factor, basic fibroblast growth factor, platelet-derived growth factor, or transforming growth factor alpha, or LPS can dramatically augment the secretion of IL-6 by RPE cells. Thus, these inflammatory mediators can act alone or synergistically with IFN-gamma to activate RPE cells and dramatically increase the expression and secretion of IL-6. In contrast, IL-1 was not detected following stimulation with any of the above-mentioned cytokines or LPS. Characterization of IL-6 protein production by RPE cells revealed that 98% of the protein is promptly secreted by the cell, its induction is dependent upon the time and concentration of the stimulant, and the continuous presence of the stimulant is required for IL-6 production. Moreover, Western blot (immunoblot) analysis of secreted proteins revealed that IL-6 was produced in multiple molecular forms.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Synergistic effects of gamma interferon on inflammatory mediators that induce interleukin-6 gene expression and secretion by human retinal pigment epithelial cells. 855 3

The purpose of this study was to determine if certain growth factors and bacterial products induce pleural mesothelial cells (PMC) to produce nitric oxide (NO). Confluent monolayers of rat PMC were exposed to epidermal growth factor (EGF), platelet-derived growth factor (PDGF), or lipopolysaccharide (LPS) individually and in various combinations for 24-72 h. Concentrations of nitrite and nitrate were quantified and used as an indirect measure of NO production. LPS stimulation resulted in a significant increase in nitrite/nitrate concentration, but neither EGF nor PDGF alone or combined had any significant effect relative to control. However, LPS combined with either EGF or PDGF caused a significant increase in nitrite/nitrate concentration relative to LPS alone and growth factor alone. The highest level level of nitrite/nitrate concentration was observed with the triple combination of LPS, EGF, and PDGF. Nitrite/nitrate accumulation was significantly increased at 24 h by all combinations, and continued to increase, with the highest concentration observed after 72 h of exposure. Nitrite/nitrate production was significantly inhibited by NG-nitro-L-arginine methyl ester and this inhibition was reversed by the addition of L-arginine, suggesting that nitrite and nitrate were derived from the L-arginine-dependent formation of NO. These data indicate that PMC can be induced to produce relatively large amounts of NO in response to growth factors combined with LPS.
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PMID:Nitric oxide synthesis by rat pleural mesothelial cells: induction by growth factors and lipopolysaccharide. 855 91

We have examined basal and lipopolysaccharide (LPS)-induced release of epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), growth-regulated peptide alpha (GRO alpha), leukaemia inhibitory factor (LIF), macrophage inflammatory protein-1a (MIP-1 alpha) and platelet-derived growth factor-AB (PDGF-AB) in peripheral blood mononuclear cells (PBMC) from 20 persons with either high (n = 10) or low (n = 10) levels of high-density lipoprotein (HDL). PBMC were incubated with 100 ng LPS/ml for up to 160 h, and showed a significantly higher release of the chemokines GRO alpha (P = 0.04) and MIP-1 alpha (P < 0.01) in persons with high HDL, whereas levels of GM-CSF were similar. Levels of EGF, LIF and PDGF-AB were always low, and remained unaltered during 160 h of incubation. These findings indicate that PBMC from persons with high or low levels of HDL have different functional properties, of importance in cell recruitment and activation.
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PMID:LPS-induced release of EGF, GM-CSF, GRO alpha, LIF, MIP-1 alpha and PDGF-AB in PBMC from persons with high or low levels of HDL lipoprotein. 858 Mar 73

Tranilast has been reported to reduce restenosis rate after angioplasty, but its mechanism is still unclear. We investigated the effect of tranilast against platelet-derived growth factor (PDGF) in PDGF's proliferative effect and PDGF's inhibitory effect on cytokine-induced nitric oxide (NO) production in vascular smooth muscle cells (VSMC). NO production was measured by Griess reaction. NO synthase (NOS) protein was evaluated by Western blot with monoclonal anti-rat inducible NOS antibody. A combination of interleukin-1 beta (IL-1 beta 1 ng/ml), tumor necrosis factor-alpha (TNF-alpha 2,000 U/ml), and lipopolysaccharide (100 ng/ml) significantly increased NO production and NOS protein, and tranilast significantly enhanced both in a dose-dependent manner. PDGF (100 ng/ml) significantly reduced both cytokine-induced NO production and NOS protein induction, but tranilast completely abolished these inhibitory effects. In the presence of cytokines, serum-stimulated cell proliferation was significantly inhibited by cytokine-induced NO, whereas PDGF-stimulated proliferation was not. On the other hand, tranilast not only inhibited the proliferative effect of PDGF directly, but also restored cytokine-induced NO production and its antiproliferative effect in the presence of PDGF.
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PMID:Tranilast restores cytokine-induced nitric oxide production against platelet-derived growth factor in vascular smooth muscle cells. 885 74

Quiescent and non-quiescent human gingival fibroblasts (HGF) were incubated for 24 hours with Actinobacillus actinomycetemcomitans lipopolysaccharide (LPS) and/or growth factors (interleukin-1 beta [IL-1 beta], insulin, epidermal growth factor [EGF], platelet-derived growth factor [PDGF], fibroblast growth factor [FGF], and transforming growth factor-beta [TGF-beta]) to examine the ability of LPS to modify HGF proliferation in response to these autocrine and paracrine growth factors. A. actinomycetemcomitans LPS at high concentrations (> or = 9 micrograms/well) generally resulted in a reduction in DNA synthesis in quiescent and non-quiescent fibroblasts; however, LPS at low concentrations (< 9 micrograms/well) showed a minimal enhancement of DNA synthesis (40 to 60%) in quiescent and non-quiescent cells. HGF co-incubated with mitogenic agents and LPS (9 micrograms/well) exhibited suppression of growth factor-induced 3H-Tdr uptake compared to growth factor-stimulated controls. In contrast, 3H-Tdr uptake was slightly elevated with addition of LPS at low concentrations (0.09 microgram/well). These trends were seen with all growth factors tested. Non-quiescent cells, in general, were more responsive to the growth factors and LPS/growth factor combinations when compared to the quiescent HGF. HGF were further tested for the ability of LPS to alter growth factor responsiveness by pretreating the cells with LPS prior to incubation of the growth factor, as well as, subsequent addition of LPS to growth factor-pretreated cells. Similar patterns were observed as above, except IL-1 beta-pretreated quiescent and non-quiescent HGF followed by LPS addition demonstrated a marked elevation in proliferation when compared to IL-1 beta stimulated controls. These findings suggest that LPS may potentially modulate the proliferative rate of connective tissue undergoing inflammatory or growth factor-induced reparative processes in periodontal lesions.
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PMID:The effect of lipopolysaccharide on growth factor-induced mitogenesis in human gingival fibroblasts. 899 73

Glomerulosclerosis is the final outcome of a number of different causes of glomerular injury, during which the structures of the glomerulus are obliterated by extracellular matrix. Accumulating evidence suggests that infiltrating macrophages play a pivotal role in the progression to glomerulosclerosis. The present study defines the role played by macrophages at both cellular and molecular levels in the initiation of the sclerotic process in cultured rat mesangial cells. Macrophage-conditioned medium (MPCM) generated from thioglycollate-elicited, lipopolysaccharide-stimulated macrophages upregulated mesangial cell fibronectin production in a dose- and time-dependent manner, independently of cell proliferation. Immunoprecipitation of metabolically labeled 35S-fibronectin confirmed that the matrix protein was synthesized de novo. The genes for fibronectin and the matrix proteins laminin and collagen IV were also found to be upregulated 2.86 +/- 0.24-, 4.94 +/- 0.17-, and 3.03 +/- 0.31-fold over controls, respectively (P < 0.001). Macrophage modulation of matrix turnover was suggested by an upregulation of both transin and tissue inhibitor of metalloproteinase-1 gene transcription. Transforming growth factor (TGF) beta1, platelet-derived growth factor, tumor necrosis factor (TNF) alpha, or interleukin (IL)-1beta could not be detected in the MPCM per se; however, TGFbeta1 and platelet-derived growth factor AB were found to be secreted into mesangial cell culture supernatants. Secretion was augmented 1.69 +/- 0.16- and 2.28 +/- 0.28-fold, respectively (both P < 0.001), in response to MPCM. Northern blot analysis demonstrated that protein secretion had been preceded by upregulation of the genes for these cytokines (2.2 +/- 0.4-fold [P < 0.001] and 5.7 +/- 1.2-fold [P < 0.004], respectively). Incubation of MPCM with either neutralizing antibody or the growth factor receptor antagonist suramin demonstrated that TGFbeta1 played a significant, although minor, role in MPCM-stimulated fibronectin production. In conclusion, this study provides compelling evidence for a direct role of macrophages in the progression to glomerulosclerosis.
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PMID:Macrophages promote prosclerotic responses in cultured rat mesangial cells: a mechanism for the initiation of glomerulosclerosis. 933 80

During vascular injury, such as observed in atherosclerosis, restenosis, vasculitides, transplantation, or sepsis, vascular smooth muscle cells (SMC) can be exposed to platelets or platelet products. Under these conditions proliferation or cytokine production of SMC stimulated by platelets or platelet products may contribute to regulation of vascular pathogenesis. Thus, we investigated interleukin-6 (IL-6) and IL-8 production as well as proliferation of SMC in response to platelets or platelet lysates. Platelets not already preactivated by thrombin induced IL-6 (10- to 50-fold) or IL-8 production of unstimulated SMC in a cell number dependent fashion. Preactivation of platelets with thrombin potently increased the platelet-mediated IL-6 (50- to 1,000-fold) and IL-8 production of SMC. Hirudin specifically inhibited the activation of platelets with thrombin. Isolated platelets cultured in the absence of SMC did not contain detectable IL-6 or IL-8. Prestimulation (4 hours) of SMC with pathophysiologically relevant substances (lipopolysaccharide [LPS], tumor necrosis factor-alpha [TNF-alpha], or IL-1alpha) further increased the platelet-induced cytokine production. The platelet-derived SMC stimulatory activity was IL-1, since IL-1 receptor antagonist (IL-1-Ra) inhibited the platelet-induced cytokine production of SMC. Anti-platelet-derived growth factor (PDGF)-antibody did not further reduce this activity. Thrombin itself stimulated expression of IL-6 and IL-8 to some degree and induced IL-6 production of SMC synergistically with IL-1. Platelets also induced proliferation of SMC, however, anti-PDGF antibodies, rather than IL-1-Ra blocked this response. These data show that platelet-derived IL-1 stimulates cytokine production of vascular smooth muscle cells, indicating that platelet-derived IL-1 may contribute to regulation of local pathogenesis in the vessel wall by activation of the cytokine regulatory network.
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PMID:Platelet-derived interleukin-1 induces cytokine production, but not proliferation of human vascular smooth muscle cells. 941 77

Quantitation of mRNA expression by endothelial cells in vivo has been limited to larger animals from which sufficient amounts of RNA could be isolated for Northern blot analysis. In the present study, we established a technique to isolate endothelial RNA from rat aortas using en face preparations. This RNA was not contaminated with RNA from smooth muscle cells as demonstrated by the absence of smooth muscle alpha-actin RNA. Following lipopolysaccharide (LPS) administration to rats, quantitation of platelet-derived growth factor (PDGF) ligand and receptor mRNA expression was carried out by competitive reverse transcriptase-polymerase chain reaction and normalized to glyceraldehyde-3 phosphate dehydrogenase. The results of the competitive reverse transcriptase-polymerase chain reaction were compared with those obtained by en face in situ hybridization. Aortic endothelium showed a 140-fold increase in PDGF-A mRNA expression 4 hours after LPS injection. Expression levels of this growth factor declined to near base line levels within 36 hours of the LPS injection. A 52-fold increase in PDGF-B mRNA was seen at 12 hours after LPS injection but expression levels were approximately 300-fold lower than for PDGF-A. These data indicate that changes in PDGF expression by endothelium in vivo can greatly exceed those observed in cultured cells. This method should permit study of endothelial gene regulation in a variety of pathological conditions in vivo.
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PMID:A quantitative method for determination of endothelial mRNA expression in vivo: induction of platelet-derived growth factor by endotoxin. 954 51


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