Poly(ADP-ribose) synthetase activity during bleomycin-induced lung fibrosis in hamsters

doi:10.1016/0014-4800(85)90037-1

Experimental and Molecular Pathology

Volume 43, Issue 2, October 1985, Pages 162-176

https://doi.org/10.1016/0014-4800(85)90037-1 Get rights and content

Abstract

Bleomycin damages cellular DNA and is a potent inducer of pulmonary fibrosis. It has been shown to act through a superoxide-mediated mechanism. We are interested in determining the biochemical mechanisms involved in fibrosis and in this preliminary study we have examined the temporal relationship between early biochemical events associated with DNA damage and fibrosis, in lungs of hamsters after administration of 0.75 unit of bleomycin. The activities of poly(ADP-ribose) synthetase, an enzyme associated with DNA repair, inducible superoxide dismutase (SOD) and prolyl hydroxylase as well as the tissue levels of NAD⁺ and hydroxyproline in the lung were determined. All three enzyme activities expressed as per milligram DNA or per lung, increased upon bleomycin treatment over the saline-administered controls. Lung poly(ADP-ribose) synthetase activity which is sensitive to DNA breaks, increased first (24% over control in 1 day, P < 0.0001), attained the maximum value on the 5th day (952% over control, P < 0.0001), and started to decline thereafter and approached near the control value on 14th day. Bleomycin treatment induced a rapid change in the level of lung NAD⁺. After 1 day the level of NAD⁺ was reduced by 42% compared to the control (P < 0.001), further declined to 65% (P < 0.001) on the 3rd day, and stayed at that level until the 7th day. On the 14th day, however, the NAD⁺ level was still lower (29%, P < 0.05) but approaching the value in the control animals. The activity of prolyl hydroxylase showed significant increase on the 3rd day (50% over control, P < 0.0001) after bleomycin administration. The enzyme activity continued to increase until the end of the experiment (490% of control, P < 0.0001, on Day 14). The content of undialyzable hydroxyproline, a marker for collagen, was also increased significantly in the lung tissue on the 3rd day (30% over control, P < 0.05), continued to increase and reached the highest level on the 14th day (71% over control, P < 0.001). A significant increase in the activity of SOD (19% over control, P < 0.001) was seen on the 5th day which continued to increase and attained the highest value on Day 14 (115% over control, P < 0.0001). These data indicate that poly(ADP-ribose) synthesis in the lung is increased upon bleomycin administration and suggest that the toxicity of the drug in vivo is mediated through damage to cellular DNA which initiates a DNA repair response in the lung.

References (71)

C. Bernofsky et al.
An improved cycling assay for nicotinamide adenine dinucleotide
Anal. Biochem.
(1973)
R.S. Bhatnagar et al.
Evidence for free radical involvement in the hydroxylation of proline: Inhibitory by nitro blue tetrazolium
FEBS Lett.
(1972)
R.S. Bhatnagar et al.
Hydralazine-induced disturbances in collagen biosynthesis
Biochim. Biophys. Acta
(1972)
K. Brawn et al.
DNA strand scission by enzymically generated oxygen radicals
Arch. Biochem. Biophys.
(1981)
R.M. Burger et al.
Effects of O₂ on the reactions of activated bleomycin
J. Biol. Chem.
(1982)
Q.P. Ghani et al.
Oxygen enhances in vivo myocardial synthesis of poly(ADP-ribose)
Biochem. Biophys. Res. Commun.
(1978)
S.N. Giri et al.
Effects of intratracheal administration of bleomycin on GSH-shuttle enzymes, catalase, lipid peroxidation, and collagen content in the lungs of hamsters
Toxicol. Appl. Pharmacol.
(1983)
S.N. Giri et al.
Increases in lung prolyl hydroxylase and superoxide dismutase activities during bleomycin-induced lung fibrosis in hamsters
Exp. Mol. Pathol.
(1983)
A.P. Grollman et al.
Interactions of bleomycin with DNA
S. Hasegawa et al.
The polymerization of adenosine 5′-diphosphate-ribose moiety of NAD by nuclear enzyme. II. Properties of the reaction product
Biochim. Biophys. Acta
(1967)

M.Z. Hussain et al.

Involvement of superoxide in the paraquat-induced enhancement of lung collagen synthesis in organ culture

Biochem. Biophys. Res. Commun.

(1979)

J.J. Hutton et al.

A rapid assay for collagen proline hydroxylase

Anal. Biochem.

(1966)

R. Ishida et al.

Increased DNA chain breakage by combined action of bleomycin and superoxide radical

Biochem. Biophys. Res. Commun.

(1975)

E.L. Jacobson et al.

Pyridine nucleotide levels as a function of growth in normal and transformed 3T3 cells

Arch. Biochem. Biophys.

(1976)

J.M. McCord et al.

Superoxide dismutase: An enzymic function for erythrocuprein

J. Biol. Chem.

(1969)

E.G. Miller

Stimulation of nuclear poly(ADP-ribose) polymerase activity from HeLa cells by endonucleases

Biochim. Biophys. Acta

(1975)

L.W. Oberley et al.

The production of hydroxyl radical by bleomycin and iron (II)

FEBS Lett.

(1979)

L.O. Rodriguez et al.

Iron(11)-bleomycin: Biochemical and spectral properties in the presence of radical scavengers

Biochem. Biophys. Res. Commun.

(1982)

T.T. Sakai et al.

Bleomycin interactions with DNA: Studies on the role of the C-triminal cationic group of bleomycin A₂ in association with and degradation of DNA

Biochim. Biophys. Acta

(1983)

T. Shima et al.

Studies on Poly(ADP-ribose)

J. Biol. Chem.

(1969)

J.B. Stevens et al.

Induction of superoxide dismutase by oxygen in neonatal rat lung

J. Biol. Chem.

(1977)

H.P. Witschi

Exploitable biochemical approaches for the evaluation of toxic lung damage

Essay Toxicol.

(1975)

J.F. Woessner

The determination of hydroxyproline in tissue and protein samples containing proportions of this imino acid

Arch. Biochem. Biophys.

(1961)

K. Yoshihara et al.

Poly(ADP-ribose) synthetase-DNA interaction

A. Barikowiak et al.

The crypto-OH radical in the damage of DNA by bleomycin-Fe²⁺

Intl. J. Biochem.

(1982)

R.C. Benjamin et al.

Poly(ADP-ribose) synthesis in vitro programmed by damaged DNA

J. Biol. Chem.

(1980)

N.A. Berger et al.

Association of poly(ADP-ribose) synthesis with DNA damage and repair in normal human lymphocytes

J. Clin. Invest.

(1979)

N.A. Berger et al.

Defective poly(ADP-ribose) synthesis in xeroderma pigmentosum

Biochemistry

(1980)

R.S. Bhatnagar

R.S. Bhatnagar et al.

The role of collagen as a primary molecular site of environmental injury

R.S. Bhatnagar et al.

Effect of ozone on lung collagen biosynthesis

M. Bradley et al.

X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution

Nucleic Acid Res.

(1979)

W.J. Caspary et al.

Effect of deoxyribonucleic acid on the production of reduced oxygen by bleomycin and iron

Biochemistry

(1982)

D. Creissen et al.

Regulation of DNA ligase activity by poly(ADP-ribose)

Nature (London)

(1982)

S.T. Crooke et al.

Bleomycin: A review

J. Med.

(1976)

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^☆: This work was supported in parts by Research Grants 2R01 HL 2735-04 from NHLB1 (S.N.G.) and 1 R01 DE06622-01A1 from NIDR (M.Z.H.), Department of Health and Welfare, Washington, D.C. Correspondence should be addressed to M.Z.H., at 604-HSW, Department of Stomatology, University of California, San Francisco, Calif. 94143.

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Poly(ADP-ribose) synthetase activity during bleomycin-induced lung fibrosis in hamsters☆

Abstract

Anal. Biochem.

FEBS Lett.

Biochim. Biophys. Acta

Arch. Biochem. Biophys.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Toxicol. Appl. Pharmacol.

Exp. Mol. Pathol.

Biochim. Biophys. Acta

Biochem. Biophys. Res. Commun.

Anal. Biochem.

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

J. Biol. Chem.

Biochim. Biophys. Acta

FEBS Lett.

Biochem. Biophys. Res. Commun.

Biochim. Biophys. Acta

J. Biol. Chem.

J. Biol. Chem.

Essay Toxicol.

Arch. Biochem. Biophys.

The crypto-OH radical in the damage of DNA by bleomycin-Fe2+

Intl. J. Biochem.

Poly(ADP-ribose) synthesis in vitro programmed by damaged DNA

J. Biol. Chem.

Association of poly(ADP-ribose) synthesis with DNA damage and repair in normal human lymphocytes

J. Clin. Invest.

Defective poly(ADP-ribose) synthesis in xeroderma pigmentosum

Biochemistry

The role of collagen as a primary molecular site of environmental injury

Effect of ozone on lung collagen biosynthesis

X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution

Nucleic Acid Res.

Effect of deoxyribonucleic acid on the production of reduced oxygen by bleomycin and iron

Biochemistry

Regulation of DNA ligase activity by poly(ADP-ribose)

Nature (London)

Bleomycin: A review

J. Med.

The crypto-OH radical in the damage of DNA by bleomycin-Fe²⁺