REGULATION OF BIGLYCAN AND DECORIN SYNTHESIS BY CONNECTIVE TISSUE GROWTH FACTOR IN CULTURED BOVINE AORTIC ENDOTHELIAL CELLS :

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  • Yamamoto,Chika
    Department of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University
  • Oh-i,Mami
    Department of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University
  • Fujiwara,Yasuyuki
    Department of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University
  • Kaji,Toshiyuki
    Department of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University
  • Nishida,Takashi
    Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine and Dentistry
  • Nakanishi,Tohru
    Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine and Dentistry
  • Takigawa,Masaharu
    Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine and Dentistry
  • Kinsella,Michael G.
    Department of Pathology, School of Medicine, University of Washington
  • Wight,Thomas N.
    Department of Pathology, School of Medicine, University of Washington

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Connective tissue growth factor (CTGF) is a regulator of vascular endothelial cell functions but little is know about the regulation of proteoglycan synthesis by the growth factor. Since endothelial cell proteoglycan synthesis is often regulated depending on the cell density, dense and sparse cultures of bovine aortic endothelial cells were metabolically labeled with [^<35>S]sulfate or ^<35>S-labaled amino acids in the presence of recombinant human CTGF. The labeled proteoglycans were characterized by DEAE-Sephacel ion exchange chromatography and Sepharose CL-4B molecular sieve chromatography. The glycosaminoglycan (GAGs) M_r and composition were analyzed by Sepharose CL-6B chromatography, and the core protein M_r was analyzed by SDS-polyacrylamide gel electrophoresis, before and after digestion with papain, heparitinase or chondroitin ABC lyase. The core proteins were identified by Western blot analysis and core protein mRNAs were determined by quantitative RT-PCR. The results indicated that CTGF suppresses the synthesis of biglycan but newly induces that of decorin in endothelial cells when the cell density is low. However, neither the hydrodynamic size nor the GAG chain length of these two small chondroitin/dermatan sulfate proteoglycans was changed by the growth factor. The present data suggest that CTGF is a regulator of the synthesis of small chondroitin/dermatan sulfate proteoglycans, biglycan in vascular endothelial cells depending on the cell density.

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