Protective effect of hemin against cadmium-induced testicular damage in rats

doi:10.1016/j.tox.2008.12.022

Toxicology

Volume 257, Issue 3, 29 March 2009, Pages 153-160

https://doi.org/10.1016/j.tox.2008.12.022 Get rights and content

Abstract

The protective effect of hemin, the heme oxygenase-1 inducer, was investigated in rats with cadmium induced-testicular injury, in which oxidative stress and inflammation play a major role. Testicular damage was induced by a single i.p. injection of cadmium chloride (2 mg/kg). Hemin was given for three consecutive days (40 μmol/kg/day, s.c.), starting 1 day before cadmium administration. Hemin treatment significantly increased serum testosterone level that was reduced by cadmium. Hemin compensated deficits in the antioxidant defense mechanisms (reduced glutathione, and catalase and superoxide dismutase activities), and suppressed lipid peroxidation in testicular tissue resulted from cadmium administration. Also, hemin attenuated the cadmium-induced elevations in testicular tumor necrosis factor-α and nitric oxide levels, and caspase-3 activity. Additionally, hemin ameliorated cadmium-induced testicular tissue damage observed by light and electron microscopic examinations. The protective effect afforded by hemin was abolished by prior administration of zinc protoporphyrin-IX, the heme oxygenase-1 inhibitor. It was concluded that hemin, through its antioxidant, anti-inflammatory and antiapoptotic effects, represents a potential therapeutic option to protect the testicular tissue from the detrimental effects of cadmium.

Introduction

Cadmium is a wide spread environmental pollutant, arising primarily from battery, electroplating, pigment, plastic and fertilizer industries. Acute and chronic cadmium toxicity is associated with severe damage in various organs, particularly the testes, in both humans and animals (Lauwerys, 1979). Previous studies demonstrated a significant decrease in testicular weights with reduced androgen secretion as a result of exposure to cadmium indicating that testicular endocrine function had been compromised (Gunnarsson et al., 2003, Yang et al., 2006). Oxidative stress and inflammation are the main factors implicated in cadmium-induced tissue injury (Kayama et al., 1995, Oteiza et al., 1999, Gupta et al., 2004). Several antioxidants and anti-inflammatory agents were found to be effective in minimizing cadmium-induced organ damage, including the testes with restoration of testicular steriodogenesis (Min et al., 2002, Kara et al., 2007, Yadav and Khandelwal, 2008).

Heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme catabolism, is important for regulating the adaptive protection of tissues against oxidative stress and inflammation (Hangaishi et al., 2000, Morio et al., 2006). Hemin, the HO-1 inducer, was proved effective in protecting against oxidative and inflammatory tissue injuries in various experimental models (Yoneya et al., 2000, Wen et al., 2007, Chen et al., 2008). Recently, pharmacological induction of HO-1 by hemin protected rat testes against ischemia/reperfusion injury resulted from testicular torsion and detorsion (Yang et al., 2007). However, to the best of our knowledge, the effect of hemin against cadmium-induced testicular damage was not yet studied.

Therefore, the present study was conducted to evaluate the protective effect of hemin in testicular injury induced by acute cadmium exposure in rats. Also, the effect of zinc protoporphyrin-IX (ZnPP), the HO-1 inhibitor, on the testicular protective activity of hemin was investigated. For this purpose, serum testosterone level was measured. The oxidant-antioxidant status of testicular tissue was assessed by measuring malondialdehyde (MDA) and reduced glutathione (GSH) levels, and catalase and superoxide dismutase (SOD) activities. The levels of tumor necrosis factor-α (TNF-α) and nitric oxide (NO) in testicular tissue were determined. Also, caspase-3 activity (as an indicator of cell apoptosis) was assessed in testicular homogenates. In addition, light and electron microscopic examinations of testicular tissue were performed.

Section snippets

Animals

Male Sprague-Dawely rats, weighing 200 ± 10 g were obtained from the Animal House, College of Medicine, Al-Ahsa, King Faisal University. The animals were housed at 24 ± 1 °C, 45 ± 5% humidity and 12 h light–12 h dark cycle. They were supplied with standard laboratory chow and water ad libitum, and left to acclimatize for 1 week before the experiments. The experimental protocol was approved by the Local Animal Care Committee. The experimental procedures were carried out in accordance with international

Effect of hemin on serum testosterone level

The rats received cadmium chloride in a single i.p. dose (2 mg/kg) showed significantly lower serum testosterone level as compared to the control animals. However, the level of serum testosterone was significantly higher in animals received cadmium and treated with hemin in comparison to hemin non-treated group. The effect of hemin was completely abolished by prior administration of ZnPP (Fig. 1).

Effect of hemin on testicular oxidant-antioxidant status

Cadmium administration resulted in a significant increase in lipid peroxidation associated with

Discussion

Cadmium is an environmental and industrial pollutant that adversely affects male reproductive system in humans and animals. Increased production of reactive oxygen species and inflammatory cytokines seems to be crucial in the pathogenesis of tissue injury in response to cadmium exposure (Kayama et al., 1995, Acharya et al., 2008).

In the present study, hemin treatment was started one day before cadmium administration and continued for 3 consecutive days in order to obtain the benefits of both

Conflict of interest

There are none.

References (42)

U.R. Acharya et al.
Effects of vitamins C and E on spermatogenesis in mice exposed to cadmium
Reprod. Toxicol.
(2008)
T. Ashino et al.
Negative feedback regulation of lipopolysaccharide-induced inducible nitric oxide synthase gene expression by heme oxygenase-1 induction in macrophages
Mol. Immunol.
(2008)
E.M. Bulger et al.
Induction of heme-oxygenase 1 inhibits endothelial cell activation by endotoxin and oxidant stress
Surgery
(2003)
H. Chiu et al.
Differential induction of heme oxygenase-1 in macrophages and hepatocytes during acetaminophen-induced hepatotoxicity in the rat: effects of hemin and biliverdin
Toxicol. Appl. Pharmacol.
(2002)
D. Gunnarsson et al.
Cadmium-induced decrement of the LH receptor expression and c-AMP levels in the testis of rats
Toxicology
(2003)
R.S. Gupta et al.
Vitamin C and vitamin E protect the rat testes from cadmium-induced reactive oxygen species
Mol. Cells
(2004)
M. Hangaishi et al.
Induction of heme oxygenase-1 can act protectively against cardiac ischemia/reperfusion in vivo
Biochem. Biophys. Res. Comm.
(2000)
F. Kayama et al.
Role of tumor necrosis factor-α in cadmium-induced hepatotoxicity
Toxicol. Appl. Pharmacol.
(1995)
T. Kusakabe et al.
Changes of heavy metal, metallothionein and heat shock proteins in Sertoli cells induced by cadmium exposure
Toxicol. In Vitro
(2008)
K.-S. Min et al.
Glucocorticoids suppress the inflammation-mediated tolerance to acute toxicity of cadmium in mice
Toxicol. Appl. Pharmacol.
(2002)

L.A. Morio et al.

Hepatic expression of heme oxygenase-1 and antioxidant response element-mediated genes following administration of ethinyl estradiol to rats

Toxicol. Appl. Pharmacol.

(2006)

G.O. Noriega et al.

Effect of acetaminophen on heme metabolism in rat liver

Int. J. Biochem. Cell. Biol.

(2000)

P.I. Oteiza et al.

Camium-induced testes oxidative damage in rats can be influenced by dietary zinc intake

Toxicology

(1999)

S. Thijssen et al.

Low cadmium exposure triggers a biphasic oxidative stress response in mice kidneys

Toxicology

(2007)

T. Wen et al.

Upregulation of heme oxygenase-1 with hemin prevents d-galactosamine and lipopolysaccharide-induced acute hepatic injury in rats

Toxicology

(2007)

N. Yadav et al.

Effect of picroliv on cadmium induced testicular damage in rats

Food Chem. Toxicol.

(2008)

S. Yang et al.

The protective role of heme oxygenase-1 induction on testicular tissues after testicular torsion and detorsion

J. Urol.

(2007)

R. Yoneya et al.

Hemin pretreatment ameliorates aspects of the nephropathy induced by mercuric chloride in the rat

Toxicol. Lett.

(2000)

X. Zhang et al.

Carbon monoxide inhibition of apoptosis during ischemia-reperfusion lung injury is dependent on the p38 MAPK pathway and involves caspase-3

J. Biol. Chem.

(2003)

D.E. Baranano et al.

Biliverdin reductase: a major physiologic cytoprotectant

Proc. Natl. Acad. Sci. U.S.A.

(2002)

K. Chan et al.

An important function of Nrf2 in combating oxidative stress: detoxification of acetaminophen

Proc. Natl. Acad. Sci. U.S.A.

(2001)

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Protective effect of hemin against cadmium-induced testicular damage in rats

Abstract

Introduction

Section snippets

Animals

Effect of hemin on serum testosterone level

Effect of hemin on testicular oxidant-antioxidant status

Discussion

Conflict of interest

Reprod. Toxicol.

Mol. Immunol.

Surgery

Toxicol. Appl. Pharmacol.

Toxicology

Mol. Cells

Biochem. Biophys. Res. Comm.

Toxicol. Appl. Pharmacol.

Toxicol. In Vitro

Toxicol. Appl. Pharmacol.

Toxicol. Appl. Pharmacol.

Int. J. Biochem. Cell. Biol.

Toxicology

Toxicology

Toxicology

Food Chem. Toxicol.

J. Urol.

Toxicol. Lett.

J. Biol. Chem.

Biliverdin reductase: a major physiologic cytoprotectant

Proc. Natl. Acad. Sci. U.S.A.

An important function of Nrf2 in combating oxidative stress: detoxification of acetaminophen

Proc. Natl. Acad. Sci. U.S.A.