Thrombospondin-1 Affects Bovine Luteal Function via Transforming Growth Factor-Beta1-Dependent and Independent Actions1

ABSTRACT Thrombospondin-1 (THBS1) and transforming growth factor-beta1 (TGFB1) are specifically up-regulated by prostaglandin F2alpha in mature corpus luteum (CL). This study examined the relationship between the expression of THBS1 and TGFB1 and the underlying mechanisms of their actions in luteal endothelial cells (ECs). TGFB1 stimulated SMAD2 phosphorylation and SERPINE1 levels in dose- and time-dependent manners in luteal EC. THBS1 also elevated SERPINE1; this effect was abolished by TGFB1 receptor-1 kinase inhibitor (SB431542). The findings here further imply that THBS1 activates TGFB1 in luteal ECs: THBS1 increased the effects of latent TGFB1 on phosphorylated SMAD (phospho-SMAD) 2 and SERPINE1. THBS1 silencing significantly decreased SERPINE1 and levels of phospho-SMAD2. Lastly, THBS1 actions on SERPINE1 were inhibited by LSKL peptide (TGFB1 activation inhibitor); LSKL also counteracted latent TGFB1-induced phospho-SMAD2. We found that TGFB1 up-regulated its own mRNA levels and those of THBS1. Both compounds generated apoptosis, but THBS1 was significantly more effective (2.5-fold). Notably, this effect of THBS1 was not mediated by TGFB1. THBS1 and TGFB1 also differed in their activation of p38 mitogen-activated protein kinase. Whereas TGFB1 rapidly induced phospho-p38, THBS1 had a delayed effect. Inhibition of p38 pathway by SB203580 did not modulate TGFB1 effect on cell viability, but it amplified THBS1 actions. THBS1-stimulated caspase-3 activation coincided with p38 phosphorylation, suggesting that caspase-induced DNA damage initiated p38 phosphorylation. The in vitro data suggest that a feed-forward loop exists between THBS1, TGFB1, and SERPINE1. Indeed all these three genes were similarly induced in the regressing CL. Their gene products can promote vascular instability, apoptosis, and matrix remodeling during luteolysis.

[1]  R. Meidan,et al.  Functions and Transcriptional Regulation of Thrombospondins and Their Interrelationship with Fibroblast Growth Factor-2 in Bovine Luteal Cells1 , 2014, Biology of reproduction.

[2]  Fang Yu,et al.  Pregnancy-associated genes contribute to antiluteolytic mechanisms in ovine corpus luteum. , 2013, Physiological genomics.

[3]  David D. Roberts,et al.  Thrombospondin-1 Signaling through CD47 Inhibits Self-renewal by Regulating c-Myc and Other Stem Cell Transcription Factors , 2013, Scientific Reports.

[4]  H. Bollwein,et al.  Vascular and immune regulation of corpus luteum development, maintenance, and regression in the cow. , 2012, Domestic animal endocrinology.

[5]  R. Meidan,et al.  Regulation of Angiogenesis-Related Prostaglandin F2alpha-Induced Genes in the Bovine Corpus Luteum1 , 2012, Biology of reproduction.

[6]  A. Ghosh,et al.  PAI‐1 in tissue fibrosis , 2012, Journal of cellular physiology.

[7]  Dulce Maroni,et al.  TGFB1 disrupts the angiogenic potential of microvascular endothelial cells of the corpus luteum , 2011, Journal of Cell Science.

[8]  J. Steibel,et al.  Deciphering the luteal transcriptome: potential mechanisms mediating stage-specific luteolytic response of the corpus luteum to prostaglandin F₂α. , 2011, Physiological genomics.

[9]  Josephine C. Adams,et al.  The thrombospondins. , 2011, Cold Spring Harbor perspectives in biology.

[10]  D. Ribatti,et al.  Non-peptidic Thrombospondin-1 Mimics as Fibroblast Growth Factor-2 Inhibitors , 2010, The Journal of Biological Chemistry.

[11]  E. Lo,et al.  Neurovascular effects of CD47 signaling: Promotion of cell death, inflammation, and suppression of angiogenesis in brain endothelial cells in vitro , 2009, Journal of neuroscience research.

[12]  S. Cullen,et al.  Caspase activation pathways: some recent progress , 2009, Cell Death and Differentiation.

[13]  M. Rincón,et al.  Non-Classical P38 Map Kinase Functions: Cell Cycle Checkpoints and Survival , 2008, International journal of biological sciences.

[14]  R. Meidan,et al.  Induction of heparanase in bovine granulosa cells by luteinizing hormone: possible role during the ovulatory process. , 2009, Endocrinology.

[15]  Chao Jiang,et al.  Prostaglandin F2α Stimulates the Expression and Secretion of Transforming Growth Factor B1 Via Induction of the Early Growth Response 1 Gene (EGR1) in the Bovine Corpus Luteum , 2008 .

[16]  Marco Presta,et al.  Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain. , 2008, The international journal of biochemistry & cell biology.

[17]  B. Rueda,et al.  Prostaglandin F2alpha stimulates the expression and secretion of transforming growth factor B1 via induction of the early growth response 1 gene (EGR1) in the bovine corpus luteum. , 2008, Molecular endocrinology.

[18]  H. Kliem,et al.  Expression and localisation of extracellular matrix degrading proteases and their inhibitors during the oestrous cycle and after induced luteolysis in the bovine corpus luteum. , 2007, Reproduction.

[19]  D. Wink,et al.  CD47 Is Necessary for Inhibition of Nitric Oxide-stimulated Vascular Cell Responses by Thrombospondin-1* , 2006, Journal of Biological Chemistry.

[20]  C. Heldin,et al.  Non-Smad TGF-β signals , 2005, Journal of Cell Science.

[21]  C. Heldin,et al.  Non-Smad TGF-beta signals. , 2005, Journal of cell science.

[22]  B. Berisha,et al.  Regulation of corpus luteum function in cattle--an overview. , 2004, Reproduction in domestic animals = Zuchthygiene.

[23]  N. Buduneli,et al.  Plasminogen activators and plasminogen activator inhibitors in gingival crevicular fluid of cyclosporin A-treated patients. , 2004, Journal of clinical periodontology.

[24]  Ying E. Zhang,et al.  Smad-dependent and Smad-independent pathways in TGF-β family signalling , 2003, Nature.

[25]  W. Laug,et al.  Increased plasminogen activator inhibitor-1 in keloid fibroblasts may account for their elevated collagen accumulation in fibrin gel cultures. , 2003, The American journal of pathology.

[26]  J. Dagorn,et al.  Gene expression profiling by DNA microarray analysis in mouse embryonic fibroblasts transformed by rasV12 mutated protein and the E1A oncogene , 2003, Molecular Cancer.

[27]  E. Skrzypczak‐Jankun,et al.  A novel form of the plasminogen activator inhibitor created by cysteine mutations extends its half-life: relevance to cancer and angiogenesis. , 2003, Molecular cancer therapeutics.

[28]  R. Derynck,et al.  Smad-dependent and Smad-independent pathways in TGF-beta family signalling. , 2003, Nature.

[29]  F. Lai,et al.  Plasminogen activator inhibitor-1 polymorphism is associated with progressive renal dysfunction after acute rejection in renal transplant recipients. , 2002, Transplantation.

[30]  M. Yanagisawa,et al.  Distinct cellular localization and regulation of endothelin-1 and endothelin-converting enzyme-1 expression in the bovine corpus luteum: implications for luteolysis. , 2001, Endocrinology.

[31]  C. Stocco,et al.  Opposite Effect of Prolactin and Prostaglandin F2α on the Expression of Luteal Genes as Revealed by Rat cDNA Expression Array. , 2001, Endocrinology.

[32]  P. Bornstein,et al.  Thrombospondins as matricellular modulators of cell function. , 2001, The Journal of clinical investigation.

[33]  C. Stocco,et al.  Opposite effect of prolactin and prostaglandin F(2 alpha) on the expression of luteal genes as revealed by rat cDNA expression array. , 2001, Endocrinology.

[34]  R. Schlapbach,et al.  TGF‐β induces the expression of the FLICE‐inhibitory protein and inhibits Fas‐mediated apoptosis of microglia , 2000 .

[35]  P. Friedl,et al.  Integrin-associated protein and thrombospondin-1 as endothelial mechanosensitive death mediators. , 2000, Biochemical and biophysical research communications.

[36]  J. Murphy-Ullrich,et al.  Activation of latent TGF-β by thrombospondin-1: mechanisms and physiology , 2000 .

[37]  J. Murphy-Ullrich,et al.  Activation of latent TGF-beta by thrombospondin-1: mechanisms and physiology. , 2000, Cytokine & growth factor reviews.

[38]  J. Juengel,et al.  Mechanisms controlling the function and life span of the corpus luteum. , 2000, Physiological reviews.

[39]  P. Higgins,et al.  Growth state‐dependent regulation of plasminogen activator inhibitor type‐1 gene expression during epithelial cell stimulation by serum and transforming growth factor‐β1 , 1999, Journal of cellular physiology.

[40]  S. Schultz-Cherry,et al.  The Activation Sequence of Thrombospondin-1 Interacts with the Latency-associated Peptide to Regulate Activation of Latent Transforming Growth Factor-β* , 1999, The Journal of Biological Chemistry.

[41]  R. Diegelmann,et al.  Differential regulation of PAI-1 gene expression in human fibroblasts predisposed to a fibrotic phenotype. , 1999, Experimental cell research.

[42]  D. Clark,et al.  Transforming growth factor-beta (TGF-beta). , 1998, The international journal of biochemistry & cell biology.

[43]  R. Roberts,et al.  Control of extracellular matrix remodelling within ovarian tissues: localization and regulation of gene expression of plasminogen activator inhibitor type-1 within the ovine corpus luteum. , 1997, Journal of reproduction and fertility.

[44]  K. Skorecki,et al.  Extracellular Signal-regulated Kinase and the Small GTP-binding Protein, Rac, Contribute to the Effects of Transforming Growth Factor-β1 on Gene Expression* , 1996, The Journal of Biological Chemistry.

[45]  K. Skorecki,et al.  Extracellular signal-regulated kinase and the small GTP-binding protein, Rac, contribute to the effects of transforming growth factor-beta1 on gene expression. , 1996, The Journal of biological chemistry.

[46]  H. Krutzsch,et al.  Regulation of Transforming Growth Factor-β Activation by Discrete Sequences of Thrombospondin 1 (*) , 1995, The Journal of Biological Chemistry.