Activation of Na + /Ca 2+ Exchanger in Kinin B 1 Receptor-Stimulated Human Fibroblast Is Associated with Collagen Production

Hypertension ◽  
2000 ◽  
Vol 36 (suppl_1) ◽  
pp. 710-720
Author(s):  
Jose R Romero ◽  
Dennis A Ricupero ◽  
Alicia Rivera ◽  
Ronald H Goldstein ◽  
Paul R Conlin
Keyword(s):  
Connective Tissue ◽  
Type I Collagen ◽  
Fold Increase ◽  
Receptor Activation ◽  
Tissue Growth ◽  
Northern Analysis ◽  
Type I ◽  
Mrna Levels ◽  
Paracrine Factors ◽  
Collagen Formation

P148 The arterial wall in hypertension is characterized by thickening of the media, in part due to increased deposition of connective tissue. Autocrine and paracrine factors may participate in this process; including products of the kallikrein-kinin system. We evaluated early signal transduction events and effects on collagen formation in B 1 -stimulated human myofibroblast cells (IMR-90). We measured cytosolic calcium (Ca cyt ) levels in cells loaded with FURA-2AM. Gene expression of connective tissue growth factor (CTGF) and α1(I) collagen was determined by estimating mRNA levels using Northern analysis of B 1 stimulated cells. Activation of the B 1 receptor with des-arg 10 -kallidin stimulated a three-fold increase in CTGF mRNA by increasing its stability. Furthermore, B 1 receptor activation caused an increase in α1(I) collagen mRNA and a four-fold increase in type I collagen synthesis in these cells; events not observed in B 2 receptor-stimulated cells. Activation of the B 1 receptor stimulated a dose dependent rise in Ca cyt (EC 50 =1.9nM) which was completely inhibited by des-arg 10 -[leu 9 ]-kallidin (100nM), a B 1 receptor antagonist. Isosmotic replacement of extracellular Na + with N -methyl,D-glucamine blocked > 90% of the B 1 stimulated rise in Ca cyt . A similar effect was observed when Ca 2+ was removed from the extracellular media, suggesting a role for the plasma membrane Na + /Ca 2+ exchanger (NCX). To further define a role for the NCX on CTGF formation we used dichlorobenzamil (DCB) and KB-R7943, two specific NCX inhibitors. DCB completely blocked the activation of B 1 receptor induced increase in CTGF mRNA stability while not affecting basal CTGF mRNA levels. In contrast, preincubation with EIPA, an amiloride analog, did not affect basal or stimulated CTGF mRNA levels. Furthermore, 60μM KB-R7943 blocked the B 1 stimulated rise in Ca cyt . NCX isoform 1 was identified in these cells using RT-PCR and immuno-detection. Thus, B 1 receptor stimulation increases fibrogenesis through a mechanism that involves modulation of cation metabolism via reverse-mode activation of NCX.

Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen

2009 ◽  
Vol 60 (7) ◽  
pp. 2142-2155 ◽  
Author(s):  
Markella Ponticos ◽  
Alan M. Holmes ◽  
Xu Shi-Wen ◽  
Patricia Leoni ◽  
Korsa Khan ◽  
...  
Keyword(s):  
Growth Factor ◽  
Connective Tissue ◽  
Connective Tissue Growth Factor ◽  
Transcriptional Activation ◽  
Lung Fibrosis ◽  
Type I Collagen ◽  
Tissue Growth ◽  
Pivotal Role ◽  
Type I

scholarly journals Src Inhibition Attenuates Liver Fibrosis by Preventing Hepatic Stellate Cell Activation and Decreasing Connective Tissue Growth Factor

Cells ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 558 ◽  
Author(s):  
Hye-Young Seo ◽  
So-Hee Lee ◽  
Ji-Ha Lee ◽  
Yu Na Kang ◽  
Jae Seok Hwang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Liver Fibrosis ◽  
Connective Tissue ◽  
Connective Tissue Growth Factor ◽  
Mouse Liver ◽  
Type I Collagen ◽  
Transforming Growth Factor ◽  
Tissue Growth ◽  
Type I ◽  
Induced Expression

The SRC kinase family comprises non-receptor tyrosine kinases that are ubiquitously expressed in all cell types. Although Src is reportedly activated in pulmonary and renal fibrosis, little is known regarding its role in liver fibrosis. This study investigated whether the inhibition of Src protects against liver fibrosis. The expression of Src was upregulated in thioacetamide (TAA)-induced fibrotic mouse liver and cirrhosis of patients, and phospho-Src was upregulated during activation of hepatic stellate cells (HSC). In addition, Src inhibition reduced the expression of α-smooth muscle actin (αSMA) in primary HSCs and suppressed transforming growth factor β (TGF-β)-induced expression of connective tissue growth factor (CTGF) in hepatocytes. Src inhibitor Saracatinib also attenuated TAA-induced expression of type I collagen, αSMA, and CTGF in mouse liver tissues. The antifibrotic effect of Src inhibitors was associated with the downregulation of Smad3, but not of signal transducer and activator of transcription 3 (STAT3). In addition, Src inhibition increased autophagy flux and protected against liver fibrosis. These results suggest that Src plays an important role in liver fibrosis and that Src inhibitors could be treat liver fibrosis.

Control of type I collagen formation in the lung

1991 ◽  
Vol 261 (2) ◽  
pp. L29-L40 ◽  
Author(s):  
R. H. Goldstein
Keyword(s):  
Type I Collagen ◽  
Potential Interaction ◽  
Complex Nature ◽  
Type I ◽  
Mrna Levels ◽  
Structural Protein ◽  
Inflammatory Reactions ◽  
Collagen Formation ◽  
Collagen Accumulation

Type I collagen is a major structural protein in the lung, the accumulation of which is stimulated during certain inflammatory reactions in the lung. Accumulating evidence suggests that type I collagen formation parallels changes in steady-state mRNA levels. Specific inflammatory substances modulate transcription of collagen genes and stabilization of collagen mRNA in vitro. However, the precise role for any particular mediator during fibrotic processes is difficult to identify because of the complex nature of the inflammatory reaction and potential interaction among mediators. The signal transduction mechanisms that regulate collagen accumulation remain to be defined. This review focuses on the regulation of collagen accumulation in the lung by specific inflammatory substances.

Cyr61 and CTGF are molecular markers of bladder wall remodeling after outlet obstruction

2002 ◽  
Vol 283 (4) ◽  
pp. E765-E774 ◽  
Author(s):  
Brahim Chaqour ◽  
Catherine Whitbeck ◽  
Ji-Soo Han ◽  
Edward Macarak ◽  
Pat Horan ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Bladder Wall ◽  
Outlet Obstruction ◽  
Tissue Growth ◽  
Ribonuclease Protection Assay ◽  
Detrusor Muscle ◽  
Type I ◽  
Mrna Levels ◽  
Rat Urinary Bladder ◽  
Ribonuclease Protection

Cysteine-rich protein (Cyr61) and connective tissue growth factor (CTGF) are key immediate early growth factors with functions in cell proliferation, differentiation, and extracellular matrix synthesis. Studies were performed to assess the gene expression profile of Cyr61 and CTGF in rat urinary bladder during growth in response to partial outlet obstruction. The mRNA levels of Cyr61 as determined by ribonuclease protection assay increased sharply after 1 day and remained elevated throughout the time period of the obstruction. This correlates well with increased bladder weight. The CTGF mRNA levels seemed to peak within the second week of the urethral obstruction and correlate well with increased type I collagen mRNA. The expression pattern of either Cyr61 or CTGF proteins corroborated that of their respective mRNAs. Immunohistochemical analyses showed that immunoreactivity of Cyr61 was confined to detrusor smooth muscle and that of CTGF was detected within both detrusor muscle and lamina propria layers. These data strongly indicate the involvement of Cyr61 and CTGF in bladder wall remodeling as a result of the outlet obstruction.

scholarly journals Connective Tissue Growth Factor (IGFBP-rP2) Expression and Regulation in Cultured Bovine Endothelial Cells*

Endocrinology ◽  
1999 ◽  
Vol 140 (4) ◽  
pp. 1575-1580 ◽  
Author(s):  
Mary Boes ◽  
Brian L. Dake ◽  
Barbara A. Booth ◽  
Ngozi E. Erondu ◽  
Youngman Oh ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Connective Tissue ◽  
Proteolytic Activity ◽  
Connective Tissue Growth Factor ◽  
Tissue Growth ◽  
Large Vessel ◽  
Northern Analysis ◽  
Mrna Levels ◽  
Microvessel Endothelial Cells

Abstract Media from large vessel endothelial cells (pulmonary artery, aorta) contained intact connective tissue growth factor (CTGF) and a dominant 19-kDa band. N-terminal analysis of the 19-kDa band showed sequence corresponding to CTGF amino acid 181–190, suggesting that the 19-kDa band represented a proteolytic fragment of CTGF. Intact CTGF was increased by cAMP but not by transforming growth factor-β (TGFβ). CTGF messenger RNA (mRNA) was not changed by cAMP nor TGFβ. In two microvessel endothelial cells, mRNA was found at low levels by PCR and Northern analysis, but no CTGF protein was seen on Western analysis. In the microvessel cells, TGFβ increased and cAMP did not change CTGF mRNA levels, with neither TGFβ nor cAMP increasing CTGF protein. The discordance between protein and mRNA levels in large vessel and microvessel endothelial cells was mostly explained by the effects of cAMP and TGFβ on media proteolytic activity; in large vessel cells, cAMP inhibited degradation of CTGF, whereas in microvessel cells, TGFβ and cAMP stimulated proteolytic activity against CTGF. We conclude that in large vessel endothelial cells, cAMP increased intact CTGF protein by inhibiting degradation of CTGF, whereas TGFβ stimulated neither CTGF mRNA nor protein; in microvessel cells, TGFβ increased CTGF mRNA, while both TGFβ and cAMP stimulated CTGF degradation.

Sign in / Sign up

Export Citation Format

Share Document