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Biochem J. May 15, 1997; 324(Pt 1): 305–310.
PMCID: PMC1218431

Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target.


The addition of human recombinant interleukin-1beta (IL-1beta) to cultures of lapine articular chondrocytes provoked the synthesis of large amounts of NO and reduced the production of type-II collagen. NG-Monomethyl-l-arginine (L-NMA), an inhibitor of NO synthase, strongly suppressed the production of NO and partially relieved the inhibition of collagen synthesis in response to IL-1beta. The NO donor S-nitrosoacetylpenicillamine (SNAP), on the other hand, inhibited collagen production. IL-1 lowered the abundance of Col2A1 mRNA in an NO-independent manner. Collectively, these data indicate that IL-1 suppresses collagen synthesis at two levels: a pretranslational level which is NO-independent, and a translational or post-translational level which is NO-mediated. These effects are presumably specific as L-NMA and SNAP had no effect on total protein synthesis or on the distribution of newly synthesized proteins between the cellular and extracellular compartments. Prolyl hydroxylase is an important enzyme in the post-translational processing of collagen, and its regulation and cofactor requirements suggest possible sensitivity to NO. Extracts of cells treated with IL-1 or SNAP had lower prolyl hydroxylase activity, and L-NMA was partially able to reverse the effects of IL-1. These data suggest that prolyl hydroxylase might indeed be a target for NO. Because underhydroxylated collagen monomers fail to anneal into stable triple helices, they are degraded intracellularly. Inhibition of prolyl hydroxylase by NO might thus account for the suppressive effect of this radical on collagen synthesis.

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Selected References

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  • Stadler J, Stefanovic-Racic M, Billiar TR, Curran RD, McIntyre LA, Georgescu HI, Simmons RL, Evans CH. Articular chondrocytes synthesize nitric oxide in response to cytokines and lipopolysaccharide. J Immunol. 1991 Dec 1;147(11):3915–3920. [PubMed]
  • Palmer RM, Hickery MS, Charles IG, Moncada S, Bayliss MT. Induction of nitric oxide synthase in human chondrocytes. Biochem Biophys Res Commun. 1993 May 28;193(1):398–405. [PubMed]
  • Rediske JJ, Koehne CF, Zhang B, Lotz M. The inducible production of nitric oxide by articular cell types. Osteoarthritis Cartilage. 1994 Sep;2(3):199–206. [PubMed]
  • Evans CH, Watkins SC, Stefanović-Racić M. Nitric oxide and cartilage metabolism. Methods Enzymol. 1996;269:75–88. [PubMed]
  • Charles IG, Palmer RM, Hickery MS, Bayliss MT, Chubb AP, Hall VS, Moss DW, Moncada S. Cloning, characterization, and expression of a cDNA encoding an inducible nitric oxide synthase from the human chondrocyte. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11419–11423. [PMC free article] [PubMed]
  • Maier R, Bilbe G, Rediske J, Lotz M. Inducible nitric oxide synthase from human articular chondrocytes: cDNA cloning and analysis of mRNA expression. Biochim Biophys Acta. 1994 Sep 21;1208(1):145–150. [PubMed]
  • Stefanovic-Racic M, Stadler J, Evans CH. Nitric oxide and arthritis. Arthritis Rheum. 1993 Aug;36(8):1036–1044. [PubMed]
  • Ialenti A, Moncada S, Di Rosa M. Modulation of adjuvant arthritis by endogenous nitric oxide. Br J Pharmacol. 1993 Oct;110(2):701–706. [PMC free article] [PubMed]
  • McCartney-Francis N, Allen JB, Mizel DE, Albina JE, Xie QW, Nathan CF, Wahl SM. Suppression of arthritis by an inhibitor of nitric oxide synthase. J Exp Med. 1993 Aug 1;178(2):749–754. [PMC free article] [PubMed]
  • Stefanovic-Racic M, Meyers K, Meschter C, Coffey JW, Hoffman RA, Evans CH. N-monomethyl arginine, an inhibitor of nitric oxide synthase, suppresses the development of adjuvant arthritis in rats. Arthritis Rheum. 1994 Jul;37(7):1062–1069. [PubMed]
  • Amin AR, Di Cesare PE, Vyas P, Attur M, Tzeng E, Billiar TR, Stuchin SA, Abramson SB. The expression and regulation of nitric oxide synthase in human osteoarthritis-affected chondrocytes: evidence for up-regulated neuronal nitric oxide synthase. J Exp Med. 1995 Dec 1;182(6):2097–2102. [PMC free article] [PubMed]
  • Sakurai H, Kohsaka H, Liu MF, Higashiyama H, Hirata Y, Kanno K, Saito I, Miyasaka N. Nitric oxide production and inducible nitric oxide synthase expression in inflammatory arthritides. J Clin Invest. 1995 Nov;96(5):2357–2363. [PMC free article] [PubMed]
  • Taskiran D, Stefanovic-Racic M, Georgescu H, Evans C. Nitric oxide mediates suppression of cartilage proteoglycan synthesis by interleukin-1. Biochem Biophys Res Commun. 1994 Apr 15;200(1):142–148. [PubMed]
  • Häuselmann HJ, Oppliger L, Michel BA, Stefanovic-Racic M, Evans CH. Nitric oxide and proteoglycan biosynthesis by human articular chondrocytes in alginate culture. FEBS Lett. 1994 Oct 3;352(3):361–364. [PubMed]
  • van de Loo FA, Arntz OJ, Otterness IG, van den Berg WB. Protection against cartilage proteoglycan synthesis inhibition by antiinterleukin 1 antibodies in experimental arthritis. J Rheumatol. 1992 Mar;19(3):348–356. [PubMed]
  • Hauselmann HJ, Flechtenmacher J, Michal L, Thonar EJ, Shinmei M, Kuettner KE, Aydelotte MB. The superficial layer of human articular cartilage is more susceptible to interleukin-1-induced damage than the deeper layers. Arthritis Rheum. 1996 Mar;39(3):478–488. [PubMed]
  • Kuettner KE. Biochemistry of articular cartilage in health and disease. Clin Biochem. 1992 Jun;25(3):155–163. [PubMed]
  • Goldring MB, Birkhead J, Sandell LJ, Kimura T, Krane SM. Interleukin 1 suppresses expression of cartilage-specific types II and IX collagens and increases types I and III collagens in human chondrocytes. J Clin Invest. 1988 Dec;82(6):2026–2037. [PMC free article] [PubMed]
  • Chandrasekhar S, Harvey AK, Higginbotham JD, Horton WE. Interleukin-1-induced suppression of type II collagen gene transcription involves DNA regulatory elements. Exp Cell Res. 1990 Nov;191(1):105–114. [PubMed]
  • Green WT., Jr Behavior of articular chondrocytes in cell culture. Clin Orthop Relat Res. 1971 Mar-Apr;75:248–260. [PubMed]
  • Lefebvre V, Peeters-Joris C, Vaes G. Modulation by interleukin 1 and tumor necrosis factor alpha of production of collagenase, tissue inhibitor of metalloproteinases and collagen types in differentiated and dedifferentiated articular chondrocytes. Biochim Biophys Acta. 1990 May 22;1052(3):366–378. [PubMed]
  • Peterkofsky B, Diegelmann R. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry. 1971 Mar 16;10(6):988–994. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PMC free article] [PubMed]
  • Kivirikko KI, Myllylä R. Posttranslational enzymes in the biosynthesis of collagen: intracellular enzymes. Methods Enzymol. 1982;82(Pt A):245–304. [PubMed]
  • WOESSNER JF., Jr The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys. 1961 May;93:440–447. [PubMed]
  • Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem. 1982 Oct;126(1):131–138. [PubMed]
  • Kolpakov V, Gordon D, Kulik TJ. Nitric oxide-generating compounds inhibit total protein and collagen synthesis in cultured vascular smooth muscle cells. Circ Res. 1995 Feb;76(2):305–309. [PubMed]
  • Trachtman H, Futterweit S, Singhal P. Nitric oxide modulates the synthesis of extracellular matrix proteins in cultured rat mesangial cells. Biochem Biophys Res Commun. 1995 Feb 6;207(1):120–125. [PubMed]
  • Curran RD, Ferrari FK, Kispert PH, Stadler J, Stuehr DJ, Simmons RL, Billiar TR. Nitric oxide and nitric oxide-generating compounds inhibit hepatocyte protein synthesis. FASEB J. 1991 Apr;5(7):2085–2092. [PubMed]
  • Goldring MB. Control of collagen synthesis in human chondrocyte cultures by immune interferon and interleukin-1. J Rheumatol. 1987 May;14(Spec No):64–66. [PubMed]
  • Kivirikko KI, Myllylä R. Post-translational processing of procollagens. Ann N Y Acad Sci. 1985;460:187–201. [PubMed]
  • Prockop DJ, Kivirikko KI. Heritable diseases of collagen. N Engl J Med. 1984 Aug 9;311(6):376–386. [PubMed]
  • Chvapil M, Peng YM. Oxygen and lung fibrosis. Arch Environ Health. 1975 Nov;30(11):528–532. [PubMed]
  • Hussain MZ, Giri SN, Bhatnagar RS. Poly(ADP-ribose) synthetase activity during bleomycin-induced lung fibrosis in hamsters. Exp Mol Pathol. 1985 Oct;43(2):162–176. [PubMed]
  • Prockop DJ, Kivirikko KI, Tuderman L, Guzman NA. The biosynthesis of collagen and its disorders (first of two parts). N Engl J Med. 1979 Jul 5;301(1):13–23. [PubMed]
  • Hussain MZ, Ghani QP, Hunt TK. Inhibition of prolyl hydroxylase by poly(ADP-ribose) and phosphoribosyl-AMP. Possible role of ADP-ribosylation in intracellular prolyl hydroxylase regulation. J Biol Chem. 1989 May 15;264(14):7850–7855. [PubMed]
  • Brüne B, Dimmeler S, Molina y Vedia L, Lapetina EG. Nitric oxide: a signal for ADP-ribosylation of proteins. Life Sci. 1994;54(2):61–70. [PubMed]
  • Tzeng E, Shears LL, 2nd, Robbins PD, Pitt BR, Geller DA, Watkins SC, Simmons RL, Billiar TR. Vascular gene transfer of the human inducible nitric oxide synthase: characterization of activity and effects on myointimal hyperplasia. Mol Med. 1996 Mar;2(2):211–225. [PMC free article] [PubMed]

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