Glucose and TGFβ2 Modulate the Viability of Cultured Human Retinal Pericytes and Their VEGF Release

Purpose: Determine the effects of glucose and exogenous TGFβ2 on viability and VEGF release by human retinal pericytes (HRP). Methods: Human retinal pericytes (HRP) were cultured in 5 mM (physiologic) or high (18 mM) glucose with or without added TGFβ2. Viable cells were counted; TGFβ2 and VEGF in the conditioned media (CM) were measured by ELISA. Results: High glucose significantly reduced viable cell number and increased the levels of TGFβ2 and VEGF. TGFβ2 caused a significant dose-dependent effect on viable cell number and on the level of VEGF secreted into the CM by HRP in physiologic glucose, decreasing viable cell number, and increasing VEGF release per 1000 cells at a low concentration (0.1 ng/ml) and increasing viable cell number and decreasing VEGF release per 1000 cells at higher concentrations (1.0 and 10 ng/ml). TGFβ2 affected neither parameter in high glucose. Conclusions: Elevated glucose decreased HRP viability and modulated changes in TGFβ2 and VEGF release. This suggests a novel mechanism for HRP dropout in diabetic retinopathy.

[1]  Rui-Zhen Shi,et al.  Transforming growth factor-β1 enhanced vascular endothelial growth factor synthesis in mesenchymal stem cells , 2008 .

[2]  Dong Ho Jung,et al.  Effect of magnolol on TGF-beta1 and fibronectin expression in human retinal pigment epithelial cells under diabetic conditions. , 2007, European journal of pharmacology.

[3]  Z. Bian,et al.  Regulation of VEGF mRNA expression and protein secretion by TGF-beta2 in human retinal pigment epithelial cells. , 2007, Experimental eye research.

[4]  S-C Chang,et al.  High glucose alters proteoglycan expression and the glycosaminoglycan composition in placentas of women with gestational diabetes mellitus and in cultured trophoblasts. , 2007, Placenta.

[5]  M. Porta,et al.  Effects of mechanical stress and high glucose on pericyte proliferation, apoptosis and contractile phenotype. , 2006, Experimental eye research.

[6]  G. Lang,et al.  TGFβ induces transdifferentiation of iBREC to αSMA-expressing cells , 2006 .

[7]  John C. Lee,et al.  Bone morphogenetic protein‐4 enhances vascular endothelial growth factor secretion by human retinal pigment epithelial cells , 2006, Journal of cellular biochemistry.

[8]  A. Tsin,et al.  ARPE-19 Cell Growth and Cell Functions in Euglycemic Culture Media , 2006, Current eye research.

[9]  C. Pollock,et al.  PPARgamma agonists exert antifibrotic effects in renal tubular cells exposed to high glucose. , 2005, American journal of physiology. Renal physiology.

[10]  P. Vincent,et al.  Activation of p38 Has Opposing Effects on the Proliferation and Migration of Endothelial Cells* , 2005, Journal of Biological Chemistry.

[11]  A. Rotchford,et al.  Effect of diabetes mellitus and hyperglycemia on the proliferation of human Tenon's capsule fibroblasts: Implications for wound healing after glaucoma drainage surgery , 2005, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[12]  T. Wight,et al.  Characterization of chondroitin/dermatan sulfate proteoglycans synthesized by bovine retinal pericytes in culture. , 2004, Biological & pharmaceutical bulletin.

[13]  Yunpeng Du,et al.  Interaction between NO and COX pathways in retinal cells exposed to elevated glucose and retina of diabetic rats. , 2004, American journal of physiology. Regulatory, integrative and comparative physiology.

[14]  H. Snoeck,et al.  The Positive Regulatory Effect of TGF-β2 on Primitive Murine Hemopoietic Stem and Progenitor Cells Is Dependent on Age, Genetic Background, and Serum Factors1 , 2004, The Journal of Immunology.

[15]  Mary E. Choi,et al.  Transforming Growth Factor-β1 Stimulates Vascular Endothelial Growth Factor 164 via Mitogen-activated Protein Kinase Kinase 3-p38α and p38δ Mitogen-activated Protein Kinase-dependent Pathway in Murine Mesangial Cells* , 2004, Journal of Biological Chemistry.

[16]  Hai-yan Lin,et al.  Involvement of ERK1/2 pathway in TGF‐β1‐induced VEGF secretion in normal human cytotrophoblast cells , 2004, Molecular reproduction and development.

[17]  E. Piek,et al.  Pericyte production of cell-associated VEGF is differentiation-dependent and is associated with endothelial survival. , 2003, Developmental biology.

[18]  K. Pardhasaradhi,et al.  Transforming growth factor‐β induces expression of vascular endothelial growth factor in human retinal pigment epithelial cells: Involvement of mitogen‐activated protein kinases , 2003 .

[19]  Yi-fei Huang,et al.  [Downregulation of the pigment epithelium derived factor by hypoxia and elevated glucose concentration in cultured human retinal pigment epithelial cells]. , 2003, Zhonghua yi xue za zhi.

[20]  I. Herman,et al.  FGF-2 Antagonizes the TGF-β1-Mediated Induction of Pericyte α-Smooth Muscle Actin Expression: A Role for Myf-5 and Smad-Mediated Signaling Pathways , 2003 .

[21]  T. Lincoln,et al.  Expression of constitutively active cGMP-dependent protein kinase prevents glucose stimulation of thrombospondin 1 expression and TGF-beta activity. , 2003, Diabetes.

[22]  T. Shibata,et al.  Involvement of MAP kinases in TGF-beta-stimulated vascular endothelial growth factor synthesis in osteoblasts. , 2003, Archives of biochemistry and biophysics.

[23]  P. Casanello,et al.  Hyperglycaemia Inhibits Thymidine Incorporation and Cell Growth via Protein Kinase C, Mitogen‐Activated Protein Kinases and Nitric Oxide in Human Umbilical Vein Endothelium , 2003, Experimental physiology.

[24]  F. Pomero,et al.  Effects of protein kinase C inhibition and activation on proliferation and apoptosis of bovine retinal pericytes , 2003, Diabetologia.

[25]  U. Renner,et al.  Transforming Growth Factor-β Stimulates Vascular Endothelial Growth Factor Production by Folliculostellate Pituitary Cells , 2002 .

[26]  Joseph L Evans,et al.  Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes. , 2002, Endocrine reviews.

[27]  R. Kalluri,et al.  Effects of high glucose and TGF-beta1 on the expression of collagen IV and vascular endothelial growth factor in mouse podocytes. , 2002, Kidney international.

[28]  H. Ochiai,et al.  Higher concentration of transforming growth factor-beta in aqueous humor of glaucomatous eyes and diabetic eyes. , 2002, Japanese journal of ophthalmology.

[29]  O. Kozawa,et al.  Involvement of p38 MAP kinase in TGF‐β‐stimulated VEGF synthesis in aortic smooth muscle cells , 2001, Journal of cellular biochemistry.

[30]  D. Cha,et al.  Expression of vascular endothelial growth factor in response to high glucose in rat mesangial cells. , 2000, The Journal of endocrinology.

[31]  H. Ueno,et al.  Transforming growth factor-β1 modulates the expression of vascular endothelial growth factor by osteoblasts. , 1999, American journal of physiology. Cell physiology.

[32]  G. King,et al.  Glucose or diabetes activates p38 mitogen-activated protein kinase via different pathways. , 1999, The Journal of clinical investigation.

[33]  C. Sotozono,et al.  Transforming Growth Factor β2 in the Vitreous in Proliferative Diabetic Retinopathy , 1998 .

[34]  K. Miyazono,et al.  [Functions of the transforming growth factor-beta superfamily in eyes]. , 1997, Nippon Ganka Gakkai zasshi.

[35]  L. Lanting,et al.  Effects of high glucose on vascular endothelial growth factor expression in vascular smooth muscle cells. , 1997, The American journal of physiology.

[36]  N. Wahab,et al.  Expression of extracellular matrix molecules in human mesangial cells in response to prolonged hyperglycaemia. , 1996, The Biochemical journal.

[37]  R. Folberg,et al.  Upregulated expression of vascular endothelial growth factor in proliferative diabetic retinopathy. , 1996, The British journal of ophthalmology.

[38]  L. Aiello,et al.  Hypoxic regulation of vascular endothelial growth factor in retinal cells. , 1995, Archives of ophthalmology.

[39]  S. Jimenez,et al.  TGF-beta modulates the synthesis of proteoglycans by myocardial fibroblasts in culture. , 1995, Journal of molecular and cellular cardiology.

[40]  L. Aiello,et al.  Vascular endothelial growth factor in ocular fluid of patients with diabetic retinopathy and other retinal disorders. , 1994, The New England journal of medicine.

[41]  A Mathis,et al.  Detection of vascular endothelial growth factor messenger RNA and vascular endothelial growth factor-like activity in proliferative diabetic retinopathy. , 1994, Archives of ophthalmology.

[42]  Don H. Anderson,et al.  Transforming growth factor beta 2 is the predominant isoform in the neural retina, retinal pigment epithelium-choroid and vitreous of the monkey eye. , 1994, Experimental eye research.

[43]  D. Yue,et al.  Cell-associated proteoglycans of retinal pericytes and endothelial cells: modulation by glucose and ascorbic acid. , 1994, Microvascular research.

[44]  F. Malecaze,et al.  Vasculotropin-VEGF stimulates retinal capillary endothelial cells through an autocrine pathway. , 1994, Investigative ophthalmology & visual science.

[45]  S. Dieudonné,et al.  Effects of TGF-beta 2 on mineral resorption in cultured embryonic mouse long bones; 45Ca release and osteoclast differentiation and migration. , 1994, European journal of orthodontics.

[46]  J. Folkman,et al.  Synthesis and secretion of vascular permeability factor/vascular endothelial growth factor by human retinal pigment epithelial cells. , 1993, Biochemical and biophysical research communications.

[47]  L. Orci,et al.  Biphasic Effect of Transforming Growth Factor-β1 on in Vitro Angiogenesis , 1993 .

[48]  E. Ruoslahti,et al.  Negative regulation of transforming growth factor-β by the proteoglycan decorin , 1990, Nature.

[49]  H. Moses,et al.  The cell biology of transforming growth factor β , 1990 .

[50]  E. Ruoslahti,et al.  Transforming growth factor-β regulates production of proteoglycans by mesangial cells , 1990 .

[51]  G. Striker,et al.  Transforming growth factor-beta. Murine glomerular receptors and responses of isolated glomerular cells. , 1989, The Journal of clinical investigation.

[52]  R. Majack,et al.  Vascular smooth muscle cells express distinct transforming growth factor-beta receptor phenotypes as a function of cell density in culture. , 1989, The Journal of biological chemistry.

[53]  S. D'souza,et al.  Effects of transforming growth factors β1 and β2 on a mouse clonal, osteoblastlike cell line MC3T3‐E1 , 1989 .

[54]  R. Majack Beta-type transforming growth factor specifies organizational behavior in vascular smooth muscle cell cultures , 1987, The Journal of cell biology.

[55]  I. Herman,et al.  Microvascular pericytes contain muscle and nonmuscle actins , 1985, The Journal of cell biology.

[56]  J. Gitlin,et al.  Culture of retinal capillary cells using selective growth media. , 1983, Microvascular research.

[57]  Rui-Zhen Shi,et al.  Transforming growth factor-beta1 enhanced vascular endothelial growth factor synthesis in mesenchymal stem cells. , 2008, Biochemical and biophysical research communications.

[58]  Afshin Samali,et al.  Distinct effects of high-glucose conditions on endothelial cells of macrovascular and microvascular origins. , 2006, Endothelium : journal of endothelial cell research.

[59]  K. Pardhasaradhi,et al.  Transforming growth factor-beta induces expression of vascular endothelial growth factor in human retinal pigment epithelial cells: involvement of mitogen-activated protein kinases. , 2003, Journal of cellular physiology.

[60]  I. Herman,et al.  FGF-2 antagonizes the TGF-beta1-mediated induction of pericyte alpha-smooth muscle actin expression: a role for myf-5 and Smad-mediated signaling pathways. , 2003, Investigative ophthalmology & visual science.

[61]  Richard W. Light,et al.  Transforming Growth Factor β Induces Vascular Endothelial Growth Factor Elaboration from Pleural Mesothelial Cells in Vivo and in Vitro , 2002 .

[62]  P. Thompson,et al.  Transforming growth factor beta induces vascular endothelial growth factor elaboration from pleural mesothelial cells in vivo and in vitro. , 2002, American journal of respiratory and critical care medicine.

[63]  N. Ferrara,et al.  Vascular endothelial growth factor and the regulation of angiogenesis. , 2000, Recent progress in hormone research.

[64]  M. Longaker,et al.  Transforming growth factor- b 1 modulates the expression of vascular endothelial growth factor by osteoblasts , 1999 .

[65]  C. Sotozono,et al.  Transforming growth factor beta2 in the vitreous in proliferative diabetic retinopathy. , 1998, Archives of ophthalmology.

[66]  L. Orci,et al.  Biphasic effect of transforming growth factor-beta 1 on in vitro angiogenesis. , 1993, Experimental cell research.

[67]  E. Ruoslahti,et al.  Negative regulation of transforming growth factor-beta by the proteoglycan decorin. , 1990, Nature.

[68]  W. Li,et al.  Intramural pericyte degeneration in early diabetic retinopathy study in vitro. , 1990, Chinese medical journal.

[69]  H. Moses,et al.  The cell biology of transforming growth factor beta. , 1990, Biochimica et biophysica acta.

[70]  E. Ruoslahti,et al.  Transforming growth factor-beta regulates production of proteoglycans by mesangial cells. , 1990, Kidney international.

[71]  S. D'souza,et al.  Effects of transforming growth factors beta 1 and beta 2 on a mouse clonal, osteoblastlike cell line MC3T3-E1. , 1989, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[72]  M. Sporn,et al.  Transforming growth factor beta. , 1988, Advances in cancer research.

[73]  M. Sporn,et al.  Type beta transforming growth factor: a bifunctional regulator of cellular growth. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[74]  N. Wahab,et al.  J Am Soc Nephrol 11: 1607–1619, 2000 The Decorin High Glucose Response Element and Mechanism of Its Activation in Human Mesangial Cells , 2022 .