Effects Of Diisononyl Phthalate Research Articles

Diisononyl phthalate negatively perturbs testicular energy metabolism and histoarchitecture of rats

The testes undertake high-energy demanding processes like spermatogenesis and steroidogenesis as energy metabolism enzymes are essential for testicular cells. If the energy metabolism of testis cells is interrupted, the cells may be harmed. Using cellular respiration enzymes as indicators, the effect of Diisononyl phthalate (DINP) on testicular energy transduction was evaluated. Eighteen albino rats were split into three groups, each with six rats: Tween-80 (control) was administered to Group A, DINP (20 mg/kg/BW) to Group B, and 200 mg/kg DINP to Group C orally for 14 days. The testes were removed from the rats after they were sacrificed. The activities of glycolytic and mitochondrial metabolizing enzymes in the testis, as well as histological alterations, were measured. Except for hexokinase, the glycolytic and tricarboxylic acid cycle enzymes assayed were inhibited (p<0.05) in a dose-dependent manner. In addition, all respiratory chain complexes (Complex I-IV) decreased significantly (p<0.05) relative to control. The metabolic abnormalities were confirmed by histological changes in the testes. Finally, DINP exposure impairs testicular energy transduction enzymes, implying testicular cells have a reduced supply of energy for morphological and physiological functions, as the testes require a constant supply of energy for spermatogenesis and steroidogenesis processes.

Diisononyl phthalate inhibits cardiac glycolysis and oxidative phosphorylation by down-regulating cytosolic and mitochondrial energy metabolizing enzymes in murine model

The recent increase of diisononyl phthalate (DiNP) applications in a wide range of plastic consumer products, as well as its relative environmental exposure and interactions, has compelled an investigation of its toxicity. Phthalate exposure has been associated with heart dysfunction in animals in a few studies, the bulk of which are linked to the high molecular weight phthalates. The effect of DiNP on cardiac energy transduction was assessed utilizing cellular respiration enzymes as indices. Eighteen wistar rats were divided into 3 groups of six rats each: Group A received Tween-80 (control), DiNP (20 mg/kg/BW) was given to Group B, and 200mg/kg DiNP was given to Group C orally (gavage) for 14 days. The activity of cardiac glycolytic, oxidative phosphorylation enzymes and histopathological changes were assessed. The glycolytic and tricarboxylic acid cycle enzymes studied were predominantly down-regulated in a dose-dependent manner, with the exception of cardiac citrate synthase, which showed no significant variation in activity when compared to the control. Furthermore, all respiratory chain complexes (Complex I-IV) decreased significantly relative to control, with the exception of complex IV activity at 20mg/kg/BW which showed no significant difference (P<0.05). Cardiac histopathological alterations confirmed the aforementioned metabolic disturbances. Finally, DiNP exposure impairs cardiac energy transduction enzymes, implying cardiac cells produced insufficient energy (ATP) to carry out its morphological and physiological functions efficiently as the heart requires a constant supply of energy in the form of ATP to support contraction, relaxation, and prevent cardiomyopathies.

Potential roles of oxidative stress and insulin resistance in diisononyl phthalate induced dyslipidemia and hepatosteatosis in BALB/c mice

Exposure to plastic-derived endocrine disrupting chemicals (EDCs) such as phthalates have become a source of concern to human health. Phthalates are applied industrially as plasticizers in the production of various plastic products. They have been reported for the various forms of toxic responses in both human and animal studies. Diisononyl phthalate (DiNP) was recently used to replace di(2-ethylhexyl) phthalate (DEHP) due to some toxicity concerns. However, recent evidences suggest that DiNP may disrupt the endocrine system, alter lipid metabolism and induce hepatic steatosis. Hence, this work was designed to probe the potential impacts of DiNP on oxidative stress biomarkers and insulin resistance as a possible link to dyslipidemia and development of hepatic steatosis. To achieve these, twenty seven (27) male BALB/c mice were distributed into three (3) experimental groups of nine (9) mice each. Group I was the control while mice in groups II and III were exposed to 20 and 200 mg/kg body weight, (bw) DiNP respectively orally (per os, p.o.) for 28 days. Thereafter, plasma insulin, glucose, plasma lipid levels as well as insulin resistance (IR) index were determined. The effects of DiNP on hepatic biomarker of inflammation (tumor necrosis factor alpha, TNF-α), oxidative stress (malondialdehyde, MDA), antioxidants (glutathione peroxidase, reduced glutathione and catalase) were also investigated in addition to liver histopathology. Data obtained show that DiNP especially at 200 mg/kg bw significantly (p < 0.05) alter plasma glucose and lipid profile and induced IR. Other responses observed at this dose level include significant inflammation, oxidative stress and hepatic fatty degeneration (as shown in hematoxylin and eosin stained liver sections). Current findings suggest that insulin resistance and oxidative stress may influence DiNP-induced dyslipidemia and hepatic steatosis in mice.

The effect of di-isononyl phthalate on human reproductive function and in the experiment

<h3>ЦЕЛЬ РАБОТЫ</h3> Обобщить и проанализировать данные зарубежной литературы об изучении влияния диизононилфталата (ДИНФ) на репродуктивную функцию человека и животных. За последние годы производство и использование ДИНФ увеличилось, что сопровождается повышенным риском воздействия на человека, которое отражается в быстром повышении концентрации метаболитов ДИНФ, обнаруживаемых в биологических образцах людей. Исходя из результатов исследований, можно было бы заключить, что ни пре-, ни перинатальное воздействие ДИНФ не оказывает существенного влияния на женскую репродукцию. Тем не менее доказано определенное негативное влияние ДИНФ на репродуктивные параметры грызунов. Даже кратковременное воздействие ДИНФ приводило к пожизненным последствиям. У людей вопрос о том, повлечет ли за собой краткосрочное воздействие ДИНФ аналогичные последствия, окончательно не изучен. Основываясь на ограниченной информации, мы знаем, что у женщин с более высокими концентрациями ДИНФ в крови увеличивалось время, необходимое для зачатия. На многие вопросы еще предстоит ответить, например, существует ли взаимосвязь между воздействием ДИНФ и потерей беременности. Более того, точный механизм влияния ДИНФ на репродуктивные органы также не ясен и должен быть изучен в дальнейшем. Исследования на самцах животных показали, что ДИНФ обладает антиандрогенным действием. Исследования на людях также показывают, что влияние ДИНФ может приводить к изменению уровней половых гормонов и качества спермы. На сегодняшний день некоторые последствия ДИНФ все еще являются спорными. Негативное и дозозависимое влияние ДИНФ на самцов в животных моделях подтверждено, но негативные эффекты у людей до сих пор не изучены. Необходимы дальнейшие исследования для оценки последствий влияния ДИНФ на организм взрослых мужчин. Кроме того, следует определить молекулярные механизмы, посредством которых ДИНФ действует на репродуктивные органы мужчин.

The Impact of Di-Isononyl Phthalate Exposure on Specialized Epithelial Cells in the Colon.

Di-isononyl phthalate (DiNP) is a high-molecular-weight phthalate commonly used as a plasticizer for polyvinyl chloride and other end products, such as medical devices and construction materials. Most of our initial exposure to DiNP occurs by ingestion of DiNP-contaminated foods. However, little is known about the effects of DiNP on the colon. Therefore, the goal of this study was to test the hypothesis that DiNP exposure alters immune responses and impacts specialized epithelial cells in the colon. To test this hypothesis, adult female mice were orally dosed with corn-oil vehicle control or doses of DiNP ranging from 20 µg/kg/d to 200 mg/kg/d for 10-14 days. After the dosing period, mice were euthanized in diestrus, and colon tissues and sera were collected for histological, genomic, and proteomic analysis of various immune factors and specialized epithelial cells. Subacute exposure to DiNP significantly increased protein levels of Ki67 and MUC2, expression of a Paneth cell marker (Lyz1), and estradiol levels in sera compared with control. Gene expression of mucins (Muc1, Muc2, Muc3a, and Muc4), Toll-like receptors (Tlr4 and Tlr5), and specialized epithelial cells (ChgA, Lgr5, Cd24a, and Vil1) were not significantly different between treatment groups and control. Cytokine levels of IL-1RA and CXCL12 were also not significantly different between DiNP treatment groups and control. These data reveal that DiNP exposure increases circulating estradiol levels and gene expression in specialized epithelial cells with immune response capabilities (eg, goblet and Paneth cells) in the mouse colon, which may initiate immune responses to prevent further damage in the colon.

Effects of Di-Isononyl Phthalate (DiNP) on Follicular Atresia in Zebrafish Ovary.

Di-isononyl phthalate (DiNP) is a plasticizer reported to elicit hormone-like activity and disrupt metabolism and reproduction in fish and other vertebrates. In general, phthalates have been used at high concentrations beyond reported environmental levels to assess their adverse effects on fish gonadal physiology. The present study exposed adult female zebrafish to a wide range of DiNP concentrations [0.42 µg L−1 (10−9 M), 4.2 µg L−1 (10−8 M), and 42 µg L−1 (10−7 M)] for 21 days. We evaluated gene expression profiles related to apoptosis, autophagy, and oxidative stress; DNA fragmentation (TUNEL assay: terminal deoxynucleotidyl transferase dUTP nick end labeling) and caspase activity (CAS3) were also examined. Exposure to 0.42 and 4.2 µg L−1 upregulated the genes coding for tnfa and baxa, sod1, prkaa1, respectively. CAS3 immunohistochemistry revealed a higher number of positive vitellogenic oocytes in ovaries exposed to 0.42 µg L−1. Subsequently, we examined the relationship between CAS3 signaling and DNA fragmentation. Accordingly, DNA fragmentation was observed in vitellogenic follicles of fish exposed to 0.42 and 4.2 μg L−1. Our results demonstrate that follicular atresia can occur after exposure to environmental levels of DiNP for 21 days, which may adversely affect the reproductive performance of female zebrafish in a non-monotonic manner.

Exposure evaluation of diisononyl phthalate in the adults of Drosophila melanogaster: Potential risks in fertility, lifespan, behavior, and modes of action

Diisononyl phthalate (DINP) as a phthalate plasticizer is widely used in daily life and production, which shows endocrine disruption effects and has several adverse effects on the normal physiological function. Here, the effects of DINP (0.1%, 0.2%, 0.5%, and 1.0%) (v/v) on the fertility, lifespan, climbing behavior, anti-starvation ability of Drosophila melanogaster and the potential modes of action were investigated. The results showed that DINP impaired fertility in a dose-dependent manner and smaller ovarian volume, lower hatching rate, and fewer offspring was observed at higher concentrations. The effect of DINP on the lifespan showed gender-specific, and mortality was increased after exposure above 0.2% DINP. The climbing ability increased at 0.1% DINP compared with the vehicle group, while it manifested a dose-dependent decrease at higher concentrations. The anti-starvation ability exhibited hormesis after short-term culture and reduced as culture time extending. By measuring the redox status (catalase (CAT) and reactive oxygen species (ROS)) of adult flies after two exposure methods, it was found that DINP induced redox instability, which may explain the above effects at the molecular level. This study provides data to support a comprehensive analysis of DINP potential toxicity and to guide its rational use and management better.

Effects of diisononyl phthalate (DiNP) on the endocannabinoid and reproductive systems of male gilthead sea bream (Sparus aurata) during the spawning season.

Diisononyl phthalate (DiNP) is a plasticizer used to improve plastic performance in a large variety of items whichhas been reported asan endocrine-disrupting chemical (EDC) in several organisms. The endocannabinoid system (ECS) is a cellular signaling system, whose functionality is tightly involved with reproductive function. The aim of the present study was the assessment of the effects of DiNP on the gonadal ECS and on the reproductive function of male gilthead seabream Sparus aurata, an important marine aquacultured species in Europe, during the reproductive season. Fish were fed for 21 days with two diets contaminated with different nominal concentrations of DiNP (DiNP LOW at 15µg DiNP kg-1 bw day-1 and DiNP HIGH at 1500µg DiNP kg-1 bw day-1), based on the tolerable daily intake (TDI) ruled by the European Food Safety Authority for humans. The transcription of several genes related to the ECS was affected by the DiNP. Specifically, DiNP reduced the levels of endocannabinoids and endocannabinoid-like mediators, concomitant with the increase of fatty acid amide hydrolase (FAAH) activity. At the histological level, DiNP LOW induced the highest occurrence of individuals with regressed testes. Steroidogenesis was affected significantly, since plasma 11-ketotestosterone (11-KT), the main active androgen in fish, was significantly decreased by the DiNP HIGH treatment, while plasma 17β-estradiol (E2) levels were raised, associated with an increase of the gonadosomatic index (GSI). Additionally, the level of testosterone (T) was significantly increased in the DiNP LOW group, however, the same DiNP concentration reduced the levels of 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P). The production of sperm was in general not affected, since spermiation index, sperm density, survival and the duration of forward motility did not exhibit any changes compared to controls. However, computer-assisted sperm analysis (CASA) showed that DiNP reduced the percentage of motile cells. The results clearly suggest a negative effect of DiNP via the diet on the male endocrine system of gilthead sea bream during the reproductive season.

Disruption of the gonadal endocannabinoid system in zebrafish exposed to diisononyl phthalate

DiNP (Di-isononyl phthalate) has been recently introduced as DEHP (Bis (2-ethylhexyl) phthalate) substitute and due to its chemical properties, DiNP is commonly used in a large variety of plastic items. The endocannabinoid system (ECS) is a lipid signaling system involved in a plethora of physiological pathways including the control of the reproductive and metabolic processes. In this study, the effects of DiNP on the ECS of zebrafish (male and female) gonads were analyzed. Adult zebrafish were chronically exposed for 21 days via water to 3 environmentally relevant concentrations of DiNP (42 μg/L; 4.2 μg/L; 0.42 μg/L). In females, the Gonadosomatic Index (GSI) and the number of fertilized eggs were reduced by the lowest concentration of DiNP tested. The levels of two endocannabinoids, Anandamide (AEA) and 2-Arachidonoylglycerol (2-AG), were not affected, while a reduction of the N-oleoyl-ethanolamine (OEA) level was observed. Transcriptional changes were reported in relation to genes coding for the ECS receptors and the enzymes involved in the ECS pathway. DiNP exposure in males reduced the GSI as well as changed the levels of endocannabinoids. Moreover, DiNP treatment induced significative changes in the genes coding for the ECS receptors and enzymes, and significantly increased the activity of the fatty acid amide hydrolase (FAAH). In summary, in zebrafish, exposure to environmentally relevant concentrations of DiNP disrupted the ECS and affected reproduction in a gender specific manner.

PECULIARITIES OF THE TOXIC EFFECT OF DIISONONYL PHTHALATE PLASTICIZER PHTHALATE ON EXPERIMENTAL MODELS

Introduction. The purpose of the study was to determine the characteristics of the biological effect of diisononyl phthalate, a new plasticizer for polymer products. Material and methods. Toxic properties of the compound have been studied in various ways of its entering in the organism of laboratory animals, local irritating, cumulative and skin-resorptive actions have been established, the potential ability of the compound to induce remote effects in experimental models in vivo/in vitro has been revealed. Results and discussion. The obtained data show diisononyl phthalate do not pose a risk of the acute poisoning under intragastric, intraperitoneal, epicutaneous and inhalation modes of exposure, being incapable of inducing signs of the irritation of the skin and mucous membranes, is not allergic. With subchronic intragastric administration of diisononyl phthalate in doses from 10,000 to 100 mg/kg, the dose dependence of toxic effect and its ability to cause chronic polytropic poisoning of the action in the form of disturbance of the pattern of peripheral blood, calcium and phosphorus metabolism and changes in the functional state of internal organs are revealed. In the study of the reproductive toxicity against the background of the administration of the drug at a dose of 10,000 mg/kg, an increase in total postimplantation, embryonic and the postnatal mortality rate was established. Intragastric administration of diisononyl phthalate at the mentioned dose to female animals during pregnancy triggered the formation of multiple (combined), external and internal malformations of embryos that were single in response to a reduced dose of up to 100 mg/kg. Changes in the state of the reproductive function of males have not been established. In the study of mutagenic toxicity in the Ames test, the effect of the compound has not been established to lead to an increase in the number of revertant colonies, which indicates the lack of ability of diisononyl phthalate to induce point mutations. Conclusion. The results of the studies showed that further study of the toxic effect of diisononylphthalate in chronic exposure and in vitro tests is necessary.

Effects of diisononyl phthalate on osteopenia in intact mice

Osteopenia is characterized by bone loss and deterioration of trabecular bone, which leads to osteoporotic fractures. This disease is highly prevalent in industrialized areas and is associated with exposure to endocrine disrupting chemicals (EDCs). Diisononyl phthalate (DINP) is one of these EDCs and is mainly used as a plasticizer in flexible polyvinyl chloride (PVC) products. Although it is well known that exposure to DINP is harmful to humans, no studies have been reported concerning its contribution to osteopenia. Therefore, in this study, we injected DINP (2, 20, and 200mg/kg) into C3H/HeN mice for 6weeks and found that the uterus weight, bone (femur and tibia) weight, and bone length of the DINP-exposed mice were reduced compared to those of the SHAM group. On the other hand, body weight, the serum alkaline phosphatase (ALP) and inorganic phosphorus (IP) levels in the DINP treated mice were increased compared with those of the SHAM group. The tartrate-resistant acid phosphatase (TRAP) activity (bone resorption marker) was increased and the bone alkaline phosphatase (BALP) activity was lowered by the treatment with DINP as compared with the SHAM group. Furthermore, the microarchitecture of the femur and tibia in the intact mice was destroyed by the DINP injection. The tissue volume (TV), bone volume (BV), BV/TV, bone surface (BS), BS/TV, trabecular thickness (Tb.Th), and trabecular number (Tb.N) were reduced and the trabecular pattern factor (Tb.Pf), structure model index (SMI), and trabecular separation (Tb.Sp) were increased by the DINP injection. The bone mineral density (BMD) of the femur and tibia was lower in the DINP group than in the SHAM group. These results indicate that DINP contributes to an increased risk of osteopenia via destruction of the microarchitecture and enhancement of osteoclast activity.

Diisononyl phthalate induces asthma via modulation of Th1/Th2 equilibrium

Diisononyl phthalate (DINP), a member of the phthalate family, is used to plasticize polyvinyl chloride (PVC). This chemical is known to enhance airway inflammation in the OVA-induced asthma model (adjuvant effects) and aggravate allergic dermatitis. Moreover, DINP enhances the production of interleukin-4 in activated CD4+ T cells. However, the effect of DINP itself on the differentiation of naïve CD4+ T cells into T helper cells (Th1/Th2) in vitro and allergic asthma in vivo has not yet been studied. In this study, DINP was shown to suppress the polarization of Th1 and enhance the polarization of Th2 from naïve CD4+ T cells in vitro. Also, DINP induced allergic asthma via the production of IL-4, IL-5, IgE and IgG1 and the reduction of IFN-γ and IgG2a. Finally, we confirmed that exposure to DINP induces the infiltration of inflammatory cells and PAS positive cells and increases the expression of caspase-1 and caspase-3 in asthmatic mice. In conclusion, we suggest that DINP as an environmental pollutant or endocrine disruptor (ECD) induces asthma via the modulation of the Th1/Th2 equilibrium and production of Th2 mediated cytokines and immunoglobulin.

Effects of Diisononyl Phthalate on Atopic Dermatitis in Vivo and Immunologic Responses in Vitro

BackgroundDiisononyl phthalate (DINP), a principal plasticizer in many polyvinyl chloride products, has been shown to have an adjuvant effect on immunoglobulin (Ig) production in mice. However, the effects of DINP on allergic diseases have not been fully elucidated.ObjectivesIn the present study we investigated the effects of DINP on atopic dermatitis (AD)-like skin lesions induced by Dermatophagoides pteronyssinus (Dp) in atopic-prone NC/Nga mice.MethodsMice were injected intradermally with Dp on their ears and were exposed to DINP (0, 0.15, 1.5, 15, or 150 mg/kg/day) intraperitoneally. We evaluated clinical scores, ear thickening, histologic findings, protein expression of cytokines/chemokines in the ear, and serum levels of Ig and histamine. Furthermore, we investigated the effects of DINP on bone-marrow–derived dendritic cells (BMDCs) or splenocytes in vitro. After exposure to DINP (0–100 μM), cells were evaluated for phenotype and function.ResultsDINP aggravated AD-like skin lesions related to Dp. The aggravation was consistent with eosinophilic inflammation, mast cell degranulation, and thymic stromal lymphopoietin (TSLP) expression in the ear. DINP enhanced the expression of cell surface activation markers on BMDCs and their production of TARC/CCL17 (thymus- and activation-regulated chemokine) and MDC/CCL22 (macrophage-derived chemokine), as well as their capacity to stimulate Dp-specific T-cell proliferation. DINP also enhanced interleukin-4 production and Dp-stimulated proliferation of splenocytes.ConclusionsDINP can aggravate AD-like skin lesions related to Dp. The mechanisms of the aggravation might be mediated, at least partly, through the TSLP-related activation of dendritic cells and by direct or indirect activation of the immune cells.

Effects of di-isononyl phthalate, di-2-ethylhexyl phthalate, and clofibrate in cynomolgus monkeys.

The effects of the peroxisome proliferators di-isononyl phthalate (DINP) and di-2-ethylhexyl phthalate (DEHP) were evaluated in young adult male cynomolgus monkeys after 14 days of treatment, with emphasis on detecting hepatic and other effects seen in rats and mice after treatment with high doses of phthalates. Groups of 4 monkeys received DINP (500 mg/kg/day), DEHP (500 mg/kg/day), or vehicle (0.5% methyl cellulose, 10 ml/kg) by intragastric intubation for 14 consecutive days. Clofibrate (250 mg/kg/day), a hypolipidemic drug used for cholesterol reduction in human patients was used as a reference substance. None of the test substances had any effect on body weight or liver weights. Histopathological examination of tissues from these animals revealed no distinctive treatment-related effects in the liver, kidney, or testes. There were also no changes in any of the hepatic markers for peroxisomal proliferation, including peroxisomal beta-oxidation (PBOX) or replicative DNA synthesis. Additionally, in situ dye transfer studies using fresh liver slices revealed that DINP, DEHP, and clofibrate had no effect on gap junctional intercellular communication (GJIC). None of the test substances produced any toxicologically important changes in urinalysis, hematology, or clinical chemistry; however, clofibrate produced some emesis, small increases in serum triglyceride, decreased calcium, and decreased weights of testes/epididymides and thyroid/parathyroid. The toxicological significance of these small changes is questionable. The absence of observable hepatic effects in monkeys at doses that produce hepatic effects in rodents suggests that DINP, DEHP, and clofibrate would also not elicit in primates other effects such as liver cancer. These data, along with results from in vitro hepatocyte studies, indicate that rodents are not good animal models for predicting the hepatic effects of phthalates in primates, including humans.