scholarly journals Interleukin-1 β regulation of fibroblast proteoglycan synthesis involves a decrease in versican steady-state mRNA levels

1993 ◽  
Vol 294 (2) ◽  
pp. 613-620 ◽  
Author(s):  
E E Qwarnström ◽  
H T Järveläinen ◽  
M G Kinsella ◽  
C O Ostberg ◽  
L J Sandell ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Steady State ◽  
Chondroitin Sulphate ◽  
Interleukin 1 ◽  
Mrna Level ◽  
Three Dimensional ◽  
Proteoglycan Synthesis ◽  
Mrna Levels ◽  
Pronounced Decrease

This study investigates the effects of interleukin (IL)-1 beta on proteoglycan metabolism by fibroblasts surrounded by endogenous extracellular matrix. In both three-dimensional matrix cultures and long-term monolayer cultures IL-1 beta caused a significant decrease in synthesis and deposition of sulphated proteoglycans, but had no effect on release of deposited material. The decrease in synthesis became successively more pronounced, and corresponded to 40-60% of the control after 72 h incubation. The reduction was almost totally accounted for by an effect on the chondroitin ABC-lyase-sensitive proteoglycans. Gel electrophoresis showed a significant decrease in a high-molecular-mass chondroitin ABC-lyase-sensitive proteoglycan after incubation with IL-1 beta. Northern-blot analyses of total RNA revealed a pronounced decrease in the steady-state mRNA levels of versican, the large chondroitin sulphate, with levels corresponding to 10-30% of controls. In comparison, the steady-state mRNA level for decorin, the major sulphated proteoglycan synthesized by the cells, was only slightly affected. The prominent decrease in synthesis of sulphated proteoglycans induced in long-term fibroblast cultures, including the pronounced decrease in versican steady-state mRNA levels, is likely to have a significant effect on the structure of the extracellular matrix. Induction of this type of change may constitute a significant mechanism whereby IL-1 beta can affect the properties of connective tissue during inflammation and wound healing.

scholarly journals Differential modulation of degradative and repair responses of interleukin-1-treated chondrocytes by platelet-derived growth factor

1993 ◽  
Vol 292 (1) ◽  
pp. 129-136 ◽  
Author(s):  
A K Harvey ◽  
S T Stack ◽  
S Chandrasekhar
Keyword(s):  
Extracellular Matrix ◽  
Growth Factor ◽  
Articular Chondrocytes ◽  
Interleukin 1 ◽  
Receptor Expression ◽  
Platelet Derived Growth Factor ◽  
Proteinase Activity ◽  
Proteoglycan Synthesis ◽  
Mrna Levels ◽  
The Matrix

Interleukin 1 (IL-1) plays a dual role in cartilage matrix degeneration by promoting extracellular proteinase action such as the matrix metalloproteinases (increased degradation) and by suppressing the synthesis of extracellular matrix molecules (inhibition of repair). Platelet-derived growth factor (PDGF) is a wound-healing hormone which is released along with IL-1 during the inflammatory response. Since previous studies have shown that PDGF enhances IL-1 alpha effects on metalloproteinase activity, in this report, we have examined whether PDGF modifies IL-1 beta effects on cartilage proteoglycan synthesis. Initially, we confirmed that rabbit articular chondrocytes treated with IL-1 beta + PDGF induced higher proteinase activity, in comparison with IL-1-treated cells. We further observed that the increased proteinase activity correlated with an increase in the synthesis of collagenase/stromelysin proteins and a corresponding increase in the steady-state mRNA levels for both the enzymes. Studies on IL-1 receptor expression suggested that PDGF caused an increase in IL-1 receptor expression which, by augmenting the IL-1 response, may have led to the increase in proteinase induction. Analysis of proteoglycan synthesis confirmed that IL-1 reduced the incorporation of sulphated proteoglycan, aggrecan, into the extracellular matrix of chondrocytes, whereas PDGF stimulated it. However, cells treated with IL-1 + PDGF synthesized normal levels of aggrecan. This is in contrast with cells treated with IL-1 + fibroblast growth factor, in which case only proteinase activity was potentiated. The results allow us to conclude that (a) the two effector functions that play a role in matrix remodelling, namely matrix lysis (proteinase induction) and matrix repair (proteoglycan synthesis), occur via distinct pathways and (b) PDGF may play a crucial role in cartilage repair by initially causing matrix degradation followed by promoting new matrix synthesis.

Cyclic AMP analogs and retinoic acid influence the expression of retinoic acid receptor alpha, beta, and gamma mRNAs in F9 teratocarcinoma cells

1990 ◽  
Vol 10 (1) ◽  
pp. 391-396
Author(s):  
L Hu ◽  
L J Gudas
Keyword(s):  
Retinoic Acid ◽  
Steady State ◽  
Cyclic Amp ◽  
Cellular Response ◽  
Retinoic Acid Receptor ◽  
Mrna Level ◽  
Mrna Levels ◽  
Protein Synthesis Inhibitors ◽  
Receptor Alpha ◽  
F9 Teratocarcinoma

Retinoic acid (RA) receptor alpha (RAR alpha) and RAR gamma steady-state mRNA levels remained relatively constant over time after the addition of RA to F9 teratocarcinoma stem cells. In contrast, the steady-state RAR beta mRNA level started to increase within 12 h after the addition of RA and reached a 20-fold-higher level by 48 h. This RA-associated RAR beta mRNA increase was not prevented by protein synthesis inhibitors but was prevented by the addition of cyclic AMP analogs. In the presence of RA, cyclic AMP analogs also greatly reduced the RAR alpha and RAR gamma mRNA levels, even though cyclic AMP analogs alone did not alter these mRNA levels. The addition of either RA or RA plus cyclic AMP analogs did not result in changes in the three RAR mRNA half-lives. These results suggest that agents which elevate the internal cyclic AMP concentration may also affect the cellular response to RA by altering the expression of the RARs.

scholarly journals The role of hydrological transience in peatland pattern formation

2013 ◽  
Vol 1 (1) ◽  
pp. 29-43 ◽  
Author(s):  
P. J. Morris ◽  
A. J. Baird ◽  
L. R. Belyea
Keyword(s):  
Steady State ◽  
Hydraulic Properties ◽  
Three Dimensional ◽  
Structural Complexity ◽  
Plant Litter ◽  
Future Climate Change ◽  
Litter Production ◽  
Landscape Models ◽  
Water Tables

Abstract. The sloping flanks of peatlands are commonly patterned with non-random, contour-parallel stripes of distinct micro-habitats such as hummocks, lawns and hollows. Patterning seems to be governed by feedbacks among peatland hydrological processes, plant micro-succession, plant litter production and peat decomposition. An improved understanding of peatland patterning may provide important insights into broader aspects of the long-term development of peatlands and their likely response to future climate change. We recreated a cellular simulation model from the literature, as well as three subtle variants of the model, to explore the controls on peatland patterning. Our models each consist of three submodels, which simulate: peatland water tables in a gridded landscape, micro-habitat dynamics in response to water-table depths, and changes in peat hydraulic properties. We found that the strength and nature of simulated patterning was highly dependent on the degree to which water tables had reached a steady state in response to hydrological inputs. Contrary to previous studies, we found that under a true steady state the models predict largely unpatterned landscapes that cycle rapidly between contrasting dry and wet states, dominated by hummocks and hollows, respectively. Realistic patterning only developed when simulated water tables were still transient. Literal interpretation of the degree of hydrological transience required for patterning suggests that the model should be discarded; however, the transient water tables appear to have inadvertently replicated an ecological memory effect that may be important to peatland patterning. Recently buried peat layers may remain hydrologically active despite no longer reflecting current vegetation patterns, thereby highlighting the potential importance of three-dimensional structural complexity in peatlands to understanding the two-dimensional surface-patterning phenomenon. The models were highly sensitive to the assumed values of peat hydraulic properties, which we take to indicate that the models are missing an important negative feedback between peat decomposition and changes in peat hydraulic properties. Understanding peatland patterning likely requires the unification of cellular landscape models such as ours with cohort-based models of long-term peatland development.

scholarly journals Altered proinsulin conversion in rat pancreatic islets exposed long-term to various glucose concentrations or interleukin-1β

2007 ◽  
Vol 192 (2) ◽  
pp. 381-387 ◽  
Author(s):  
Andreas Börjesson ◽  
Carina Carlsson
Keyword(s):  
Pancreatic Islets ◽  
Cell Function ◽  
Culture Media ◽  
Mrna Level ◽  
Interleukin 1Β ◽  
Mrna Levels ◽  
Rat Pancreatic Islets ◽  
Significant Prolongation ◽  
Β Cell Function

In order to elucidate a possible relationship between β-cell function and conversion of proinsulin to insulin, isolated rat pancreatic islets were maintained in tissue culture for 1 week at various glucose concentrations (5.6–56 mM). Studies were also conducted on islets cultured for 48 h with interleukin-1β (IL-1β). By pulse-chase labelling and immunoprecipitation, the relative contents of newly synthesized proinsulin and insulin were determined. ELISA was used to analyse insulin and proinsulin content in medium and within islets. Using real-time PCR, the mRNA levels of proinsulin converting enzymes (PC1 and PC2) were studied. Islets cultured at 56 mM glucose had an increased proportion of newly synthesized proinsulin when compared with islets cultured at 5.6 mM glucose after a 90-min chase periods, however, no difference was observed after culture at 11 and 28 mM glucose. ELISA measurements revealed that culture at increased glucose concentrations as well as islet exposure to IL-1β increased proinsulin accumulation in the culture media. The mRNA expression of PC1 was increased after culture at 11 and 28 mM glucose. Treatment for 48 h with IL-1β increased the proportion of proinsulin both at 45 and 90 min when compared with control islets. These islets also displayed a decreased mRNA level of PC1 as well as PC2. Calculations of the half-time for proinsulin demonstrated a significant prolongation after treatment with IL-1β. We conclude that a sustained functional stimulation by glucose of islets is coupled to a decreased conversion of proinsulin which is also true for islets treated with IL-1β. This may contribute to the elevated levels of proinsulin found both at the onset of type 1 diabetes as well as in type 2 diabetes.

scholarly journals Differential regulation of γ-glutamylcysteine synthetase heavy and light subunit gene expression

1997 ◽  
Vol 326 (1) ◽  
pp. 167-172 ◽  
Author(s):  
Jiaxin CAI ◽  
Zong-Zhi HUANG ◽  
Shelly C. LU
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Steady State ◽  
Mrna Level ◽  
Rat Hepatocytes ◽  
Mrna Levels ◽  
Subunit Gene ◽  
Subunit Mrna ◽  
Glutamylcysteine Synthetase ◽  
Steady State Mrna Level

γ-Glutamylcysteine synthetase (GCS) is the rate-limiting enzyme in the biosynthesis of glutathione and is composed of a heavy and a light subunit. Although the heavy subunit is enzymically active alone, the light subunit plays an important regulatory role by making the holoenzyme function more efficiently. In the current study we examined whether conditions which are known to influence gene expression of the heavy subunit also influence that of the light subunit, and the mechanisms involved. Treatment of cultured rat hepatocytes with hormones such as insulin and hydrocortisone, or plating hepatocytes under low cell density increased the steady-state mRNA level of the heavy subunit only. Treatment with diethyl maleate (DEM), buthionine sulphoximine (BSO) and t-butylhydroquinone (TBH) increased the steady state mRNA level and gene transcription rates of both subunits. These treatments share in common their ability to induce oxidative stress and activate nuclear factor κB (NF-κB). Treatment with protease inhibitors 7-amino-1-chloro-3-tosylamido-2-heptanone (TLCK) or L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK) had no influence on the basal NF-κB and GCS subunit mRNA levels, but blocked the activation of NF-κB by DEM, BSO and TBH, and the increase in GCS heavy subunit mRNA level by BSO and TBH. On the other hand, the DEM-, BSO- and TBH-induced increase in GCS light-subunit mRNA level was unaffected by TLCK and TPCK. Thus only the heavy subunit is hormonally regulated and growth sensitive, whereas both subunits are regulated by oxidative stress. Signalling through NF-κB is involved only in the oxidative-stress-mediated changes in the heavy subunit gene expression.

scholarly journals Regulation of pituitary inhibin/activin subunits and follistatin gene expression by GnRH in female rats

2011 ◽  
Vol 210 (1) ◽  
pp. 71-79 ◽  
Author(s):  
Petra Popovics ◽  
Zoltan Rekasi ◽  
Alan J Stewart ◽  
Magdolna Kovacs
Keyword(s):  
Gene Expression ◽  
Mrna Level ◽  
Inhibin B ◽  
Female Rats ◽  
Pituitary Cells ◽  
Mrna Levels ◽  
Short Term ◽  
Α Subunit ◽  
Long Term Treatment

Pituitary inhibin B, activin B, and follistatin are local regulators of FSH. Activin B is a homodimeric molecule (βB–βB), while inhibin B contains an α and a βB subunit. The regulation of gene expression of α, βB, and follistatin by local and endocrine hormones was examined in pituitaries from female rats and in perifused pituitary cells by RT-PCR. Ovariectomy (OVX) induced an elevation in the mRNA level of α and βB subunits and follistatin. Short-term (4 h) treatment of pituitary cells with GnRH decreased both the inhibin α and the inhibin/activin βB subunit mRNA levels, while long-term treatment (20 h) with 100 nM GnRH stimulated the expression of both subunits. In contrast, the mRNA level of follistatin was elevated after the short-term GnRH treatment. Long-term exposure of pituitary cells to estradiol and inhibin B suppressed the mRNA expression of βB and had no effect on the expression of α subunit and follistatin. Our results demonstrate that the increased expressions of inhibin/activin subunits and follistatin in the post-OVX period can be induced by the lack of gonadal negative feedback, resulting in a high GnRH environment in the pituitary. This study reports for the first time that GnRH administered in high doses and for a long period stimulates the gene expression of inhibin/activin subunits and thereby may contribute to the stimulatory effect of OVX on the expression of these genes.

scholarly journals Three-Dimensional Extracellular Matrix Scaffolds by Microfluidic Fabrication for Long-Term Spontaneously Contracted Cardiomyocyte Culture

2014 ◽  
Vol 20 (21-22) ◽  
pp. 2931-2941 ◽  
Author(s):  
Jeng-Chun Mei ◽  
Aden Yuan Kun Wu ◽  
Po-Chen Wu ◽  
Nai-Chen Cheng ◽  
Wei-Bor Tsai ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Cardiomyocyte Culture ◽  
Extracellular Matrix Scaffolds

scholarly journals Syk Plays a Critical Role in the Expression and Activation of IRAK1 in LPS-Treated Macrophages

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Jae Gwang Park ◽  
Young-Jin Son ◽  
Byong Chul Yoo ◽  
Woo Seok Yang ◽  
Ji Hye Kim ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Protein Tyrosine Kinase ◽  
Interleukin 1 ◽  
Mrna Level ◽  
Critical Role ◽  
Primary Response ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
Treatment Conditions ◽  
Lps Treatment

To address how interleukin-1 receptor-associated kinase 1 (IRAK1) is controlled by other enzymes activated by toll-like receptor (TLR) 4, we investigated the possibility that spleen tyrosine kinase (Syk), a protein tyrosine kinase that is activated at an earlier stage during TLR4 activation, plays a central role in regulating the functional activation of IRAK1. Indeed, we found that overexpression of myeloid differentiation primary response gene 88 (MyD88), an adaptor molecule that drives TLR signaling, induced IRAK1 expression and that piceatannol, a Syk inhibitor, successfully suppressed the MyD88-dependent upregulation of IRAK1 under LPS treatment conditions. Interestingly, in Syk-knockout RAW264.7 cells, IRAK1 activity was almost completely blocked after LPS treatment, while providing a Syk-recovery gene to the knockout cells successfully restored IRAK1 expression. According to our measurements of IRAK1 mRNA levels, the transcriptional upregulation of IRAK1 was induced by LPS treatment between 4 and 60 min, and this can be suppressed in Syk knockout cells, providing an effect similar that that seen under piceatannol treatment. The overexpression of Syk reverses this effect and leads to a significantly higher IRAK1 mRNA level. Collectively, our results strongly suggest that Syk plays a critical role in regulating both the activity and transcriptional level of IRAK1.

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