Human urinary phthalate metabolites level and main semen parameters, sperm chromatin structure, sperm aneuploidy and reproductive hormones

doi:10.1016/j.reprotox.2013.10.001

Reproductive Toxicology

Volume 42, December 2013, Pages 232-241

https://doi.org/10.1016/j.reprotox.2013.10.001 Get rights and content

Highlights

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Urinary phthalate metabolites levels are significantly related to poorer semen quality.
•
Urinary phthalate metabolites levels are significantly negatively associated with percentage of motile sperm cells.
•
Urinary phthalate metabolites levels decrease CASA parameters (VSL, VCL).
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A positive relationship was found between the urinary phthalate metabolites level and sperm aneuploidy.

Abstract

The aim of the study was to assess the association of phthalate metabolites levels in urine with semen parameters (sperm concentration, motility, morphology, CASA parameters), sperm chromatin structure, sperm aneuploidy and reproductive hormones. The study population consisted of 269 men who were attending an infertility clinic and had normal semen concentration (20–300 mln/ml) or slight oligozoospermia (15–20 mln/ml). Participants were interviewed and provided a semen sample. The phthalate metabolites were analysed in the urine using a procedure based on the LC–MS/MS method. Urinary phthalate metabolites levels were significantly associated with a decrease in sperm motility (5OH MEHP, MEHP, MINP), CASA parameters (MBP), testosterone level (MEHP) and an increase sperm DNA damage (MBP) and sperm aneuploidy (MBzP, MBP, MEHP, MEP). In view of the importance of human reproductive health and the widespread usage of phthalates, it is important to further investigate these correlations.

Introduction

Phthalates are a class of industrial chemicals that are dialkyl- or alkylarylesters of 1,2-benzenedicarboxylic acid and have been used for a variety of purposes [1]. Their industrial applications are related to the length of their ester chain. They are divided into two distinct groups, with very different applications, toxicological properties and classification. High molecular weight phthalates (e.g. di(2-ethylhexyl)phthalate (DEHP)) with alkyl chain lengths from 8 to 13 carbons are widely used as general-purpose plasticisers in polymers, primarily in polyvinyl chloride (PVC) resins [1], to make rigid PVC more flexible and useful, such as for wiring and cables. These phthalates are also used in a variety of consumer products, flooring and wall coverings, in food contact application and medical devices (bags for blood, parenteral nutrition, tubings and catheters) [2]. Low molecular weight phthalates with an alkyl chain of 2–7 carbons (e.g. diethyl phthalate (DEP), dibutyl phthalate (DBP)) are used in personal-care products, some cosmetics/fragrances, lacquers, varnishes, as solvents and as plasticisers in cellulose acetate [2], [3], [4]. As a result of their extensive use and their moderate resistance to degradation, phthalates are distributed widely in the environment [5]. Humans are exposed via multiple pathways, such as through food, water (oral), air (inhalation), and consumer products (dermal) [6]. Phthalates are rapidly metabolised to their monoesters in humans, and some of them can be further oxidised and conjugated with glucuronide before excretion in urine or feces [7], [8], [9]. Urinary phthalate metabolites have been used extensively as biomarkers of human exposure. Because phthalates are ubiquitous in daily life, the potential consequences of human exposure to phthalates have raised concerns in the general population.

Some phthalates are reproductive and developmental toxicants in laboratory animals (DBP, DEHP, diisobutyl (DIBP)) and are classified in the EU as reproductive toxicants; whereas others are not (DEP, diisononyl (DINP), diisodecyl (DIDP)) [10], [11], [12]. DEHP induced anti-androgenic action and abnormalities of the male reproductive system in prenatally exposed animals likely affecting the normal development of the testes [10], [12], [13], [14], [15]. Hypospadias and cryptorchidism, testicular injury leading to lowered sperm counts and reduced anogenital distance (AGD) as well as a decrease in testosterone biosynthesis involving damage to the Leydig cells [16], [17] have also been observed. In particular, phthalates may have endocrine-disrupting chemical properties that may affect the reproductive and developmental processes in humans. Consequently, recent studies have investigated the potential impact and toxicity of exposure to phthalates on aberrant male reproductive development [18]. Several recent epidemiological studies have addressed the male reproductive toxicity of phthalates [19], [20], [21], [22], [23], [24], [25], [26]. Most of the studies suggest that the urinary phthalate metabolites levels (at least one of the metabolites) may affect semen quality. Moreover, it has been documented that phthalate metabolites levels are associated with increased DNA damage, a lower sperm concentration and density as well as decreased motility and morphology. In addition, the urinary phthalate metabolites levels also appeared to be negatively associated with the testosterone level [27], [28], follicle-stimulating hormone FSH [19] and luteinising hormone (LH) [23].

Although several studies have explored the association between phthalate exposure and male reproductive function, none have carefully assessed semen quality parameters, sperm chromatin structure, sperm aneuploidy and level of reproductive hormones in one study. The aim of the present study is to assess the association of the urine phthalate metabolites levels with semen quality parameters (sperm concentration, motility, CASA parameters, morphology), sperm chromatin structure, sperm aneuploidy and the level of reproductive hormones (FSH, estradiol, testosterone) in adult men. According to our knowledge, this is the first such study that examines the relationship between phthalate metabolites levels and numerical chromosomal aberrations in sperm.

Section snippets

Study population

The study population initially consisted of 344 men who were attending an infertility clinic for diagnostic purposes and who had normal semen concentration of 20–300 mln/ml or slight oligozoospermia (semen concentration of 15–20 mln/ml) (WHO 1999) [29], from the study entitled “Environmental factors and male infertility”, which is the part of the “Epidemiology of reproductive hazards in Poland – multicentre study in Poland” project supported by the National Center for Research and Development in

Study population

The study population consisted of 269 men who were attending an infertility clinics for diagnostic purposes. The mean age of the men participating in this study was 32 years old. Most of them had secondary or higher education, while about 23% had a vocational education (Table 1). Most of the participants were non smokers (Table 1). The duration of the couple's infertility lasted mostly from 1 to 2 years and from 2 to 3 years. Past diseases which may have had an impact on semen quality (e.g.

Discussion

The results of the study suggest that urinary phthalate metabolites are significantly associated with a decrease in sperm motility (5OH MEHP, MEHP, MINP), CASA parameters (VSL, VCL) (MBP), the level of testosterone (MEHP) and increase the DNA fragmentation in sperm (MBP). After applying the correction for multiple comparisons the associations between the urinary 5OHMEHP and MEHP level and the percentage of motile sperm cells and the MBP level and CASA parameters were observed.

Additionally,

Funding

This study was performed under the project “Epidemiology of reproductive hazards – multicenter study in Poland” supported by National Center for Research and Development in Poland, from grant no. PBZ-MEiN-/8/2//2006; contract no. K140/P01/2007/1.2.1.2 and the project “Lifestyle factors and semen quality” financed with a grant for statutory activity IMP 10.19.

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Human urinary phthalate metabolites level and main semen parameters, sperm chromatin structure, sperm aneuploidy and reproductive hormones

Highlights

Abstract

Introduction

Section snippets

Study population

Study population

Discussion

Funding

Human body burdens of chemicals used in plastic manufacture

Philos Trans R Soc Lond B Biol Sci

Phthalates and other additives in plastics: human exposure and associated health outcomes

Philos Trans R Soc Lond B Biol Sci

Levels of seven urinary phthalate metabolites in a human reference population

Environ Health Perspect

Phthalates and human health

Occup Environ Med

New metabolites of di(2-ethylhexyl) phthalate (DEHP) in human urine and serum after single oral doses of deuterium-labelled DEHP

Arch Toxicol

A simple pharmacokinetic model to characterize exposure of Americans to di-2-ethylhexyl phthalate

J Expo Sci Environ Epidemiol

Glucuronidation patterns of common urinary and serum monoester phthalate metabolites

Arch Toxicol

Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat

Toxicol Sci

Male reproductive tract malformations in rats following gestational and lactational exposure to di-(n-butyl)-phthalate: an antiandrogenic mechanism?

Toxicol Sci

Oral toxicity of bis(2-ethylhexyl) phthalate during pregnancy and suckling in the Long-Evans rat

Food Chem Toxicol

Effects of phthalate esters on the developing reproductive tract of male rats

Hum Reprod Update

Abnormalities of sexual development in male rats with in utero and lactational exposure to the antiandrogenic plasticizer di(2-ethylhexyl)phthalate

Environ Health Perspect

Effect of in utero exposure to di(2-ethylhexyl)phthalate on rat testes

Indian J Exp Biol

The plasticizer diethylhexyl phthalate induces malformations by decreasing fetal testosterone synthesis during sexual differentiation in the male rat

Toxicol Sci

Mode of action: impaired fetal Leydig cell function – effects on male reproductive development produced by certain phthalate esters

Crit Rev Toxicol

Decrease in anogenital distance among male infants with prenatal phthalate exposure

Environ Health Perspect

Phthalate exposure and reproductive hormones in adult men

Hum Reprod

Altered semen quality in relation to urinary concentrations of phthalate monoester and oxidative metabolites

Epidemiology

Phthalate exposure and human semen parameters

Epidemiology

The relationship between environmental exposures to phthalates and DNA damage in human sperm using the neutral comet assay

Environ Health Perspect

Urinary phthalate metabolities and biomarkers of reproductive function in young men

Epidemiology

A pilot study associating urinary concentrations of phthalate metabolites and semen quality

Syst Biol Reprod Med

Correlation of phthalate exposures with semen quality

Toxicol Appl Pharmacol

Phthalate exposure and human semen quality in Shanghai: a cross-sectional study

Biomed Environ Sci

Decreased serum free testosterone in workers exposed to high levels of di-n-butyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP): a cross-sectional study in China

Environ Health Perspect

Associations between urinary metabolites of di(2-ethylhexyl)phthalate and reproductive hormones in fertile men

Int J Androl

Manual for the examination of human semen and sperm-cervical mucus interaction

The relationship between environmental exposure to phthalates and computer-aided sperm analysis motion parameters

J Androl

Predictive value of abnormal sperm morphology in in vitro fertilization

Fertil Steril