Functional expression of β2 adrenergic receptors responsible for protection against oxidative stress through promotion of glutathione synthesis after Nrf2 upregulation in undifferentiated mesenchymal C3H10T1/2 stem cells

Adrenaline is believed to play a dual role as a neurotransmitter in the central nervous system and an adrenomedullary hormone in the peripheral tissues. In contrast to accumulating evidence for the involvement in endochondral ossification, osteoblastogenesis, and osteoclastogenesis, little attention has been paid to the role of adrenergic signals in the mechanisms underlying proliferation and differentiation of mesenchymal stem cells with self‐renewal capacity and multi‐potentiality to differentiate into osteoblast, chondrocyte, adipocyte, and myocyte lineages. Expression of mRNA was seen for different adrenergic receptor (AdR) subtypes, including β2AdR, in the mesenchymal stem cell line C3H10T1/2 cells and mouse bone marrow mesenchymal stem cells before differentiation. Exposure to adrenaline not only increased cAMP formation, phosphorylation of cAMP responsive element (CRE) binding protein (CREB) on serine133 and CRE reporter activity in a manner sensitive to propranolol, but also rendered C3H10T1/2 cells resistant to the cytotoxicity of hydrogen peroxide, but not of either 2,4‐dinitirophenol or tunicamycin. Adrenaline induced a rapid but transient increase in mRNA expression of the antioxidative gene nuclear factor E2 p45‐related factor‐2 (Nrf2) along with an increase in the cystine/glutamate antiporter subunit xCT mRNA expression. Hydrogen peroxide was less cytotoxic in cells overexpressing Nrf2, moreover, while adrenaline significantly increased xCT promoter activity with an increase in endogenous glutathione levels. These results suggest that adrenaline may selectively protect mesenchymal C3H10T1/2 cells from oxidative stress through a mechanism related to the promoted biosynthesis of glutathione in association with transient Nrf2 expression after activation of β2AdR. J. Cell. Physiol. 218: 268–275, 2009. © 2008 Wiley‐Liss, Inc.

[1]  T. Ueshima,et al.  Up-regulation of per mRNA Expression by Parathyroid Hormone through a Protein Kinase A-CREB-dependent Mechanism in Chondrocytes* , 2006, Journal of Biological Chemistry.

[2]  N. Hulo,et al.  Norepinephrine induces lipolysis in beta1/beta2/beta3-adrenoceptor knockout mice. , 2005, Molecular pharmacology.

[3]  E. Hinoi,et al.  Suppression by glutamate of proliferative activity through glutathione depletion mediated by the cystine/glutamate antiporter in mesenchymal C3H10T1/2 stem cells , 2007, Journal of cellular physiology.

[4]  S. Bannai,et al.  Induction of cystine transport activity in mouse peritoneal macrophages by bacterial lipopolysaccharide. , 1995, The Biochemical journal.

[5]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[6]  Y. Yoneda,et al.  Pharmacological topics of bone metabolism: glutamate as a signal mediator in bone. , 2008, Journal of pharmacological sciences.

[7]  E. Hinoi,et al.  Functional GABA(B) receptors expressed in cultured calvarial osteoblasts. , 2002, Biochemical and biophysical research communications.

[8]  A. Togari,et al.  Effects of beta-adrenergic agonists on bone-resorbing activity in human osteoclast-like cells. , 2003, Biochimica et biophysica acta.

[9]  K. Negishi,et al.  A mechanism for glutamate toxicity in the C6 glioma cells involving inhibition of cystine uptake leading to glutathione depletion , 1992, Neuroscience.

[10]  T. Nagatsu,et al.  Expression of mRNAs for neuropeptide receptors and β-adrenergic receptors in human osteoblasts and human osteogenic sarcoma cells , 1997, Neuroscience Letters.

[11]  Jane Mitchell,et al.  β2‐adrenergic receptors expressed on murine chondrocytes stimulate cellular growth and inhibit the expression of Indian hedgehog and collagen type X , 2008, Journal of cellular biochemistry.

[12]  N. Nakamichi,et al.  Glutamate Inhibits Chondral Mineralization through Apoptotic Cell Death Mediated by Retrograde Operation of the Cystine/Glutamate Antiporter* , 2006, Journal of Biological Chemistry.

[13]  E. Wagner,et al.  The Molecular Clock Mediates Leptin-Regulated Bone Formation , 2005, Cell.

[14]  E. Hinoi,et al.  Serine racemase suppresses chondrogenic differentiation in cartilage in a Sox9‐dependent manner , 2008, Journal of cellular physiology.

[15]  Masaki Noda,et al.  Leptin regulation of bone resorption by the sympathetic nervous system and CART , 2005, Nature.

[16]  Patricia Ducy,et al.  Leptin Regulates Bone Formation via the Sympathetic Nervous System , 2002, Cell.

[17]  R. Papay,et al.  Stimulation of cyclic AMP in chondrocyte cultures: effects on sulfated‐proteoglycan synthesis , 1984, FEBS letters.

[18]  A. Friedenstein,et al.  Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers , 1987, Cell and tissue kinetics.

[19]  H. Kronenberg,et al.  Developmental regulation of the growth plate , 2003, Nature.

[20]  N. Hulo,et al.  Norepinephrine Induces Lipolysis in β1/β2/β3-Adrenoceptor Knockout Mice , 2005, Molecular Pharmacology.

[21]  O. Bagasra,et al.  Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Clifford J. Tabin,et al.  Regulation of Rate of Cartilage Differentiation by Indian Hedgehog and PTH-Related Protein , 1996, Science.

[23]  J. Price,et al.  The cell biology of bone growth. , 1994, European journal of clinical nutrition.

[24]  D. Prockop.  Marrow Stromal Cells as Stem Cells for Nonhematopoietic Tissues , 1997, Science.

[25]  M. Phillip,et al.  Endocrine Regulation of the Growth Plate , 2005, Hormone Research in Paediatrics.

[26]  G. Karsenty.  The complexities of skeletal biology , 2003, Nature.

[27]  D. Miao,et al.  Ihh enhances differentiation of CFK-2 chondrocytic cells and antagonizes PTHrP-mediated activation of PKA. , 2002, Journal of cell science.

[28]  Peter A. Jones,et al.  Multiple new phenotypes induced in 10T 1 2 and 3T3 cells treated with 5-azacytidine , 1979, Cell.

[29]  E. Hinoi,et al.  Osteoblast protects osteoclast devoid of sodium-dependent vitamin C transporters from oxidative cytotoxicity of ascorbic acid. , 2007, European journal of pharmacology.

[30]  A. McMahon,et al.  Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation. , 1999, Genes & development.