Co-expression analysis of lncRNA and mRNA identifies potential adipogenesis regulatory non-coding RNAs involved in the transgenerational effects of tributyltin.

The obesity epidemic is considered a global public health crisis, with an increase in caloric intake, sedentary lifestyles and/or genetic predispositions as contributing factors. Although the positive energy balance is one of the most significant causes of obesity, recent research has linked early exposure to Endocrine-Disrupting Chemicals (EDCs) such as the obesogen tributyltin (TBT) to the disease epidemic. In addition to their actions on the hormonal profile, EDCs can induce long-term changes in gene expression, possibly due to changes in epigenetic patterns. Long non-coding RNAs (lncRNAs) are epigenetic mediators that play important regulatory roles in several biological processes, through regulation of gene transcription and/or translation. In this study, we explored the differential expression of lncRNAs in gonadal white adipose tissue samples from adult male C57BL/6J F4 generation, female C57BL/6J offspring exposed (F0 generation) to 50 nM TBT or 0.1% DMSO (control of vehicle) via drinking water provided during pregnancy and lactation, analyzing RNA-seq data from a publicly available dataset (GSE105051). A total of 74 lncRNAs were differentially expressed (DE), 22 were up-regulated and 52 were down-regulated in the group whose F4 ancestor was exposed in utero to 50nM TBT when compared to those exposed to 0.1% DMSO (control). Regulation of DE lncRNAs and their potential partner genes in gonadal white adipose tissue of mice ancestrally exposed to EDC TBT may be related to the control of adipogenesis, as pathway enrichment analyses showed that these gene partners are mainly involved in the metabolism of lipids and glucose and in insulin-related pathways, which are essential for obesity onset and control.

Effects of microplastics (MPs) and tributyltin (TBT) alone and in combination on bile acids and gut microbiota crosstalk in mice.

Microplastics (MPs) and tributyltin (TBT) are both potential environmental pollutants that enter organisms through the food chain and affect bodily functions. However, the effects and mechanisms of MPs and TBT exposure (especially the co-exposure of both pollutants) on mammals remain unclear. In this study, Ф5µm MPs (5MP) was administered alone or in combination with TBT to investigate the health risk of oral exposure in mice. All three treatments induced inflammation in the liver, altered gut microbiota composition and disturbed fecal bile acids profiles. In addition to decreasing triglyceride (TG) and increasing aspartate aminotransferase (AST) and macrophage-expressed gene 1 (Mpeg1), 5MP induced hepatic cholestasis by stimulating the expression of the cholesterol hydroxylase enzymes CYP8B1 and CYP27A1, and inhibiting multidrug resistance-associated protein 2 and 3 (MRP2, MRP3), and bile-salt export pump (BSEP) to prevent bile acids for entering the blood and bile. Correspondingly, 5MP treatment decreased 7-ketolithocholic acid (7-ketoLCA) and taurocholic acid (TCA), which were positively correlated with decreased Bacteroides and Marvinbryantia and negatively correlated with increased Bifidobacterium. In addition, TBT increased interferon γ (IFNγ) and Mpeg1 levels to induce inflammation, accompanied by decreased 7-ketoLCA, tauro-alpha-muricholic acid (T-alpha-MCA) and alpha-muricholic acid (alpha-MCA) levels, which were negatively related to Coriobacteriaceae_UCG-002 and Bifidobacterium. Co-exposure to 5MP and TBT also decreased TG and induced bile acids accumulation in the liver due to inhibited BSEP, which might be attributed to the co-regulation of decreased T-alpha-MCA and Harryflintia. In conclusion, the administration of 5MP and TBT alone and in combination could cause gut microbiome dysbiosis and subsequently alter bile acids profiles, while the combined exposure of 5MP and TBT weakened the toxic effects of 5MP and TBT alone.

Histological study of gonadal tissues of adult Artemia salina (Linnaeus 1758) and immunohistochemistry by Caspase 3 and HSP70 to detect specific apoptosis markers on gonadal tissues after exposure to TBTCl.

Background: Several types of research have been recently carried out on the biological effects of TBTs, including investigations of genitals in invertebrates in response to exposure to TBTs in marine water. Aim: The objective of this research was to investigate the acute effects of tributyltin chloride (TBTCl) on gonads in the adult stage of Artemia salina by use normal histology and immunohistochemistry (IHC) (Caspase 3 and HSP70) to see specific apoptosis markers. Methods: After exposure of A. salina to different concentrations of TBTCl (25, 50, 100, 200, and 300 ng.l-1), 50 adult A. salina (25 male and 25 female) were selected randomly from each concentration to histologically study the gonads. The gonad tissue was sectioned (5 µm) and some slides were stained with hematoxylin and eosin and others were stained with IHC avidin-biotin complex, and were examined under a light microscope. Results: The results showed significant differences (p < 0.05) in histological lesions between different concentrations of TBTCl. The histological lesions in the testis and ovary section were undifferentiated cells, degenerating yolk globules, and follicle cells enveloping the oocyte which was then compared with control tissue, and these effects were found to be increased in females more than in males with the highest concentration of TBTCl. Immunohistochemistry (IHC) showed that positive immunostaining was observed in the testis and ovary as brownish deposits to Caspase 3 and HSP70 antibody after exposure to TBTCl, while the testis and ovary section in control tissue had no immunoreactivity to Caspase 3 and HSP70 antibody; these effects were profoundly increased with the highest concentration of TBTCl in females more than in males. Finally, the histological lesions and IHC (Caspase 3 and HSP70) revealed that the apoptosis and immune system stress of A. salina gonad tissue damage in females were more sensitive to TBTCl toxicity as compared to white males. Conclusion: In general, the present study aimed to observe the effects TBTCl on A. salina gonads by using histological sections and IHC (Caspase 3 and HSP70), which were evaluated for the first time and have been proven to possess an important function in apoptosis marker and immune system stress in Artemia. Finally, the specific mechanisms through which TBTCl affects A. salina Caspase 3 and HSP70 expression need further investigation.

Assessment of reproductive disorder (imposex) induced by tributyltins in marine gastropods.

Imposex is a genital disorder characterized by imposition of male sexual characteristics in female gastropods due to exposure to tributyltin (TBT). TBT is used as biocidal agent in antifouling paints, applied on the ship hulls and marine submerged structures such as fishing gears and buoys. In the present study bioassay experiment was carried out to determine imposex inductive and endocrine disruptive effect of TBT in two species of gastropods of genus Thais. In this experiment normal specimens of T. bufo and T. rudolphi were exposed to three different concentrations (100, 500 and 1000ngl-1) of TBTCl for four weeks in laboratory and at the end of experiment level of free testosterone and TBT body burden was estimated by radioimmunoassay and gas chromatograph coupled with a flame photometric detector respectively. In both tested species exposed to 500 and 1000ngl-1 of TBT imposex stages developed, while in 100ng l-1 and control groups showed no imposex condition. Elevation of free testosterone level in imposex females has also been observed. These observations indicate that the TBT act as potential imposex inducer and endocrine disruptor in the targeted gastropod species and these species can be used as sensitive biomonitoring tool for TBT contamination.

Levels and trends of tributyltin (TBT) and imposex in dogwhelk (Nucella lapillus) along the Norwegian coastline from 1991 to 2017.

The banning of organotin biocides, such as tributyltin (TBT), from use in marine antifouling paints is now leading to reproductive health recovery in marine gastropod populations all over the world. TBT induces so-called imposex (superimposition of male sexual characters onto females) in certain marine gastropods, such as the common dogwhelk Nucella lapillus. In this publication, the results of the Norwegian TBT and imposex monitoring in N. lapillus from the period 1991-2017 are presented. Significantly higher levels of TBT and imposex were measured in coastal areas close to shipping lanes along most of the coast prior to 2008 than afterwards. Levels started declining after restrictions were imposed on the use of TBT in all antifouling paint applications, with a total ban in 2008. In 2017, no sign of imposex was found in N. lapillus in any of the monitoring stations along the Norwegian coastline. Based on monitoring data shown herein, the importance of long-term biomonitoring and international chemical regulations, as well as the TBT and imposex story in general, are discussed.

The obesogen tributyltin induces features of polycystic ovary syndrome (PCOS): a review.

Polycystic ovary syndrome (PCOS) is a heterogeneous syndrome characterized by abnormal reproductive cycles, irregular ovulation, and hyperandrogenism. This complex disorder has its origins both within and outside the hypothalamic-pituitary-ovarian axis. Cardio-metabolic factors, such as obesity and insulin resistance, contribute to the manifestation of the PCOS phenotype. Polycystic ovary syndrome is one of the most common endocrine disorders among women of reproductive age. Growing evidence suggested an association between reproductive and metabolic features of PCOS and exposure to endocrine-disrupting chemicals (EDC), such as bisphenol A. Further, the environmental obesogen tributyltin (TBT) was shown to induce reproductive, metabolic and cardiovascular abnormalities resembling those found in women and animal models of PCOS. However, the causal link between TBT exposure and PCOS development remains unclear. The objective of this review was to summarize the most recent research findings on the potential association between TBT exposure and development of PCOS-like features in animal models and humans.

Extended imposex monitoring in N Atlantic Spain confirms punctual attainment of European environmental objectives for TBT.

Legislation in the European Union (EU) aimed at reaching by 2015 a Good Ecological Status in regard to tributyltin (TBT, the biocide used in traditional antifouling paints). With a view to check such an achievement in N Atlantic Spain, baseline monitoring of gastropod imposex (the recommended assessment tool) was extended up to that date. In Galicia (the Western part of the study area) the use of the rock snail Nucella lapillus since 1996 had shown this environmental objective to be met as soon as 2009, but new surveys reveal no further improvement thereafter. As for the Eastern Cantabrian coast, imposex levels in the mud snail Nassarius reticulatus progressively declined from 2006 to 2015, when records finally complied with expectations. Both data sets are confronted and discussed in relation to the diverse environmental factors that may be determining the distribution of gastropods in these regions.

Tributyltin chloride disrupts aortic vascular reactivity and increases reactive oxygen species production in female rats.

Organotin compounds, such as tributyltin (TBT), are environment contaminants that induce bioaccumulation and have potential toxic effects on marine species and mammals. TBT have been banned by the International Maritime Organization in 2003. However, the assessment of butyltin and metal contents in marine sediments has demonstrated high residual levels of TBT in some cases exceeding 7000 ng Sn g-1. The acceptable daily intake (ADI) level for TBT established by the World Health Organization is 0.5 µg/kg bw/day is based on genotoxicity, reproduction, teratogenicity, immunotoxicity, and mainly neurotoxicity. However, their effect on the cardiovascular system is not well understood. In this study, female rats were exposed to 0.5 µg/kg/day of TBT for 15 days with the goal of understanding the effect of TBT on vascular function. Female Wistar rats were treated daily by gavage and divided into control (n = 10) and TBT (n = 10) groups. The aortic rings were incubated with phenylephrine in both the presence and absence of endothelium. The phenylephrine concentration-response curves were generated by exposing endothelium-intact samples to NG-nitro-L-arginine methyl ester (L-NAME), apocynin, superoxide dismutase (SOD), catalase, tiron, and allopurinol. Acetylcholine (ACh) and sodium nitroprusside (SNP) were used to evaluate the relaxation response. Exposure to TBT reduced serum 17ß-estradiol E2 levels and increased vascular reactivity. After incubation with L-NAME, the vascular reactivity to phenylephrine was significantly higher. Apocynin, SOD, catalase, and tiron decreased the vascular reactivity to phenylephrine to a significantly greater extent in TBT-treated rats than in the control rat. The relaxation induced by ACh and SNP was significantly reduced in TBT rats. Exposure to TBT induced aortic wall atrophy and increased superoxide anion production and collagen deposition. These results provide evidence that exposing rats to the current ADI for TBT (0.5 µg/kg) for 15 days induced vascular dysfunction due to oxidative stress and morphological damage and should be considered an important cardiovascular risk factor.

Toxicogenomic analysis identifies the apoptotic pathway as the main cause of hepatotoxicity induced by tributyltin.

Tributyltin (TBT) is one of the most widely used organotin biocides, which has severe endocrine-disrupting effects on marine species and mammals. Given that TBT accumulates at higher levels in the liver than in any other organ, and it acts mainly as a hepatotoxic agent, it is important to clearly delineate the hepatotoxicity of TBT. However, most of the available studies on TBT have focused on observations at the cellular level, while studies at the level of genes and proteins are limited; therefore, the molecular mechanisms of TBT-induced hepatotoxicity remains largely unclear. In the present study, we applied a toxicogenomic approach to investigate the effects of TBT on gene expression in the human normal liver cell line HL7702. Gene expression profiling identified the apoptotic pathway as the major cause of hepatotoxicity induced by TBT. Flow cytometry assays confirmed that medium- and high-dose TBT treatments significantly increased the number of apoptotic cells, and more cells underwent late apoptosis in the high-dose TBT group. The genes encoding heat shock proteins (HSPs), kinases and tumor necrosis factor receptors mediated TBT-induced apoptosis. These findings revealed novel molecular mechanisms of TBT-induced hepatotoxicity, and the current microarray data may also provide clues for future studies.

Imposex and organotin compounds in ports of the Mediterranean and the Atlantic: Is the story over?

Organotin compounds are toxic substances released into the marine environment from antifouling paints. Sixty-two years following their first application and 9years after the complete ban on their use as biocides in 2003 (EC No. 782/2003), their negative impact on mollusks was still evident, as illustrated by imposex (i.e. the masculinization of female gastropods). This phenomenon is widely used to investigate tributyltin (TBT) pollution, with Hexaplex trunculus and Nassarius nitidus being considered as sensitive bioindicators. H. trunculus specimens and sediment samples collected from the ports of Cagliari (Sardinia), El Kantaoui (Tunisia) and Olhão (Portugal), along with N. nitidus individuals from the port of Faro (Portugal), were used for the assessment of imposex intensity and organotin pollution in these areas. High imposex frequency and organotin concentrations (TBT, triphenyltin (TPhT) and their degradation products) were observed, especially in Sardinia, implying that these chemicals remain a significant pollution issue in the specific Mediterranean ports. Moreover, the highly significant relationship established between penis length of imposex-affected females H. trunculus and TBT/TPhT concentrations offers a rapid and nonsacrificial proxy model for assessing the impact of organotins in coastal environments. Finally, the complementary use of more than one TBT bioindicator is advised in order to obtain more accurate results in detailed monitoring studies of TBT pollution.

Imposex and butyltin contamination still evident in Chile after TBT global ban.

Imposex in gastropods (Acanthina monodon, Oliva peruviana and Xanthochorus cassidiformis), butyltin levels in surface sediments (Coquimbo and Concepcion) and tissues (Valparaiso and Concepcion) were assessed in three areas under the influence of maritime activities along the central Chilean coast. The highest TBT concentrations were observed in São Vicente Bay (Concepcion), reaching 122.3ngSng(-1) in surface sediments and 59.7ngSng(-1) in gastropods tissue, while in Valparaiso ranged from 7.4 to 15.8ngSng(-1) in biota. The lowest TBT concentrations were detected in sediments from Coquimbo (<2ngSng(-1)), which can be attributed to a much lower ship/boat traffic (probably using TBT free products) in association to local oceanographic conditions. Despite DBT and MBT were the predominant analytes, recent inputs of TBT were evident in some areas. In fact, fishing boats may be a relevant source since they were the predominant maritime activity in the most contaminated sites. In addition, the absence of significant differences within BTs levels between both genders of A. monodon suggests that tissues from distinct sexes can be indistinctly used for future contamination studies. Imposex incidence was detected in 11 out of 15 sampled sites, indicating that environmental levels of TBT have been sufficient to induce deleterious effects on the exposed organisms. Thus, the impacts caused by TBT in Chilean coastal areas were detectable and consistent with other studies performed in South America. This present environmental contamination is probably due to the lack of regulations forbidding the use of TBT-based antifouling paints in Chile.

The Role of Epigenetics in the Latent Effects of Early Life Exposure to Obesogenic Endocrine Disrupting Chemicals.

Recent research supports a role for exposure to endocrine-disrupting chemicals (EDCs) in the global obesity epidemic. Obesogenic EDCs have the potential to inappropriately stimulate adipogenesis and fat storage, influence metabolism and energy balance and increase susceptibility to obesity. Developmental exposure to obesogenic EDCs is proposed to interfere with epigenetic programming of gene regulation, partly by activation of nuclear receptors, thereby influencing the risk of obesity later in life. The goal of this minireview is to briefly describe the epigenetic mechanisms underlying developmental plasticity and to evaluate the evidence of a mechanistic link between altered epigenetic gene regulation by early life EDC exposure and latent onset of obesity. We summarize the results of recent in vitro, in vivo, and transgenerational studies, which clearly show that the obesogenic effects of EDCs such as tributyltin, brominated diphenyl ether 47, and polycyclic aromatic hydrocarbons are mediated by the activation and associated altered methylation of peroxisome proliferator-activated receptor-γ, the master regulator of adipogenesis, or its target genes. Importantly, studies are emerging that assess the effects of EDCs on the interplay between DNA methylation and histone modifications in altered chromatin structure. These types of studies coupled with genome-wide rather than gene-specific analyses are needed to improve mechanistic understanding of epigenetic changes by EDC exposure. Current advances in the field of epigenomics have led to the first potential epigenetic markers for obesity that can be detected at birth, providing an important basis to determine the effects of developmental exposure to obesogenic EDCs in humans.

Adipogenic effects of a combination of the endocrine-disrupting compounds bisphenol A, diethylhexylphthalate, and tributyltin.

OBJECTIVE: The food contaminants bisphenol A (BPA), diethylhexylphthalate (DEHP), and tributyltin (TBT) are potent endocrine-disrupting compounds (EDC) known to interfere with adipogenesis. EDC usually act in mixtures and not as single compounds. The aim of this study was to investigate the effects of a simultaneous exposure of BPA, DEHP, and TBT on mesenchymal stem cell differentiation into adipocytes. METHODS: Multipotent murine mesenchymal stem cells (C3H10T1/2) were exposed to EDC mixtures in high concentrations, i.e. MIX-high (10 µmol/l BPA, 100 µmol/l DEHP, 100 nmol/l TBT), and in environmentally relevant concentrations, i.e. MIX-low (10 nmol/l BPA, 100 nmol/l DEHP, 1 nmol/l TBT). The exposure was performed either for the entire culture time (0-12 days) or at distinct stages of adipogenic differentiation. At day 12 of cell culture, the amount of adipocytes, triglyceride content (TG), and adipogenic marker gene expression were analyzed. RESULTS: MIX-high increased the development of adipocytes and the expression of adipogenic marker genes independently of the exposure window. The total TG amount was not increased. The low-concentrated EDC mixture had no obvious impact on adipogenesis. CONCLUSION: In EDC mixtures, the adipogenic effect of TBT and DEHP predominates single effects of BPA. Mixture effects of EDC are not deducible from single compound experiments.

Endocrine disruptors found in food contaminants enhance allergic sensitization through an oxidative stress that promotes the development of allergic airway inflammation.

In the past few decades, there has been a significant increase in incidence of allergic diseases. The hygiene hypothesis may provide some clues to explain this rising trend, but it may also be attributable to other environmental factors that exert a proallergic adjuvant effects. However, there is limited information on the risks of developing allergic asthma and related diseases through the ingestion of environmental chemicals found in food contaminants. In the present study, we have shown that oral administration of tributyltin, used as a model environmental chemical, induced oxidative-stress status in the bronchial lymph node, mesenteric lymph node and spleen, but not in the lung, where the initial step of allergic asthma pathogenesis takes place. Mice exposed to tributyltin exhibited heightened Th2 immunity to the allergen with more severe airway inflammation. Tributyltin also induced Treg cells apoptosis preferentially over non-Treg cells. All these effects of tributyltin exposure were canceled by the administration of glutathione monoethyl ester. Meanwhile, tributyltin did not affect airway inflammation of mice transferred with allergen-specific Th2 cells. Collectively, these results suggest that tributyltin exerts its pathological effect during the sensitization phase through oxidative stress that enhances the development of allergic diseases. The current study dissects the pathogenic role of oxidative stress induced by oral exposure to an environmental chemical during the sensitization phase of allergic airway inflammation and would be important for developing therapeutics for prevention of allergic diseases.

Adverse effect of antifouling compounds on the reproductive mechanisms of the ascidian Ciona intestinalis.

Fertilization and embryo development that occur in sea water are sensitive to xenobiotics from anthropogenic sources. In this work, we evaluated the influence of two antifouling biocides, tributyltin (TBT) and diuron, on the reproductive mechanisms of the marine invertebrate Ciona intestinalis. By using electrophysiological techniques, we examined the impact of these compounds on the electrical properties of the mature oocytes and of events occurring at fertilization. With different toxicity assays, we studied the effect of the two biocides on the gametes by evaluating fertilization rate and embryo development. Results show that sodium (Na⁺) currents were significantly reduced by either of the two biocides, whereas conductance was significantly increased. The fertilization current frequency and amplitude, fertilization rate and larval development were affected only by TBT. This study suggests that: (i) the two biocides affect either the electrical properties of the oocyte plasma membrane and the reproductive success representing a risk factor for the survival of the species exposed to environmental pollution; (ii) the ascidian Ciona intestinalis may represent a good model organism to test toxicity of marine pollutants. Possible mechanisms of action of the two biocides are discussed.

Imposex reduction and residual butyltin contamination in southern Brazilian harbors.

The imposex incidence was appraised in South American gastropods, considering the scenario before and after the global ban of tributyltin (TBT). A statistically significant reduction in imposex indexes was observed in Stramonita haemastoma collected in 2006 and 2010 from areas under the influence of four coastal harbors from southern Brazil. This reduction may be because of the effectiveness of the global ban issued by the International Maritime Organization, although the restrictions on TBT-based antifouling paints in Brazil might also have helped. Even so, a residual organotin contamination was still detected in female tissues (levels ranged from 7.6 to 164.9 ng Sn/g for TBT; from <2 to 214.5 ng Sn/g for dibutyltin; from <3.5 to 178.8 ng Sn/g for monobutyltin; and from <1.5 to 53 ng Sn/g for triphenyltin). Thus, although a reduction in imposex and environmental levels of organotins is expected in every ocean worldwide soon after the implementation of national and international restriction regulations, this will depend on the effectiveness of the global TBT ban; the effectiveness of local restrictions on producing, selling, and using TBT-based antifouling paints; and specific characteristics of local sediments, because metabolization rates and sorption/desorption of TBT previously deposited might affect its environmental bioavailability. Therefore, the reduction trend detected in the present study cannot be extrapolated to other Brazilian or South American coastal areas.

High tributyltin and imposex levels in the commercial muricid Thais chocolata from two Peruvian harbor areas.

The first appraisal of butyltin (BT) contamination linked with imposex incidence in Peruvian coastal areas was performed in the present study. Imposex occurrence and BT levels in female bodies were analyzed in the rock snail Thais chocolata distributed along 10 sites in areas under the influence of Callao and Chimbote Harbors. Imposex levels associated to high tributyltin (TBT) concentrations were observed in six of 10 studied sites, and the highest imposex levels were seen in gastropods collected near the harbor terminals. Tributyltin concentrations were the highest reported for gastropod mollusks (up to 662 ng Sn g(-1)) in recent studies. Although TBT-based antifouling paints have been banned globally (International Maritime Organization, 2008), the combination of high imposex and TBTs level with butyltin degradation index values indicated fresh TBT inputs in these Peruvian coastal areas. Thus, the need to implement local regulatory strategies related to the use of TBT-free antifouling paints is urgent, as is a thorough assessment of the effectiveness of the TBT global ban. Moreover, the wide distribution of T. chocolata combined with its capability to elicit imposex and bioaccumulate BTs renders this a good sentinel species for TBT contamination along the western South American coast.

[Endocrine disruptors and obesity: obesogens]. / Disruptores endocrinos y obesidad: obesógenos.

Incidence and prevalence of owerweight and obesity have greatly increased over the past three decades in almost all countries around the world. This phenomenon is not easily explained by lifestyle changes in populations with very different initial habits. This has led to consider the influence of other factors, the so-called endocrine disruptors, and more specifically obesogens. This study reviewed the available evidence about polluting chemical substances which may potentially be obesogens in humans: DES, genistein, bisphenol A, organotins (TBT, TPT), and phthalates. The first three groups of substances mainly act upon estrogen receptors, while organotins and phthalates activate PPARγ. It was concluded that evidence exists of the obesogenic effect of these chemical substances in tissues and experimental animals, but few data are available in humans.

C-Phycocyanin: an effective protective agent against thymic atrophy by tributyltin.

Spirulina platensis, used worldwide as a food supplement, is a natural source of protein, vitamins, carbohydrates and polyunsaturated fatty acids. C-Phycocyanin (C-Pc), its major biliprotein, is known to possess anti-oxidant, anti-inflammatory and radical scavenging properties. Our present study showed that treatment with C-Pc protects the rats from Tributyltin (TBT) induced thymic atrophy. The results reveal TBT-induced oxidative stress mediated apoptosis in rat thymocytes in vivo and its attenuation by C-Pc. This ameliorative effect could be attributed to antioxidant activity of the biliprotein. C-Pc also increased TBTC reduced thymic weight and cellularity as well. TBTC-induced ROS generation and lowered GSH levels were restored by C-Pc, suggesting its radical scavenging properties. The various apoptotic determinants such as mitochondrial membrane potential, Bax/Bcl-2 ratio, caspase-3 activity and apoptotic cell population were effectively modulated by C-Pc treatment. We make this first observation to illustrate the effectiveness of C-Pc in reducing TBTC-induced thymic atrophy. The morphology of thymic tissue was restored to near normal by this biliprotein. The present study, therefore, suggests that C-Pc could serve as an effective natural antioxidant for efficient management of TBTC induced oxidative damage.

[Preferential induction of apoptosis in regulatory T cells by tributyltin: possible involvement in the exacerbation of allergic diseases].

OBJECTIVES: Tributyltin (TBT) has been recognized as a particularly important pollutant. Human exposure to TBT persists via consumption of TBT-containing meat and fish products. Although it is well known that high-dose TBT exerts immunotoxic effects such as thymic atrophy, the effect of low-dose TBT exposure on immune responses remains elusive. Our previous studies demonstrated that TBT at environmentally relevant doses promoted T helper (Th)2 polarization via enhancement of Th2 differentiation and preferential induction of apoptosis in Th1, which is associated with the exacerbation of Th2-driven allergic airway inflammation. In the present study, we explored the possibility that TBT might preferentially induce apoptosis in Foxp3(+) regulatory T cells (Treg), which play an indisputable role in the negative regulation of immune responses. METHODS: We established several independent Treg and Th2 clones and their susceptibilities to TBT-induced apoptosis were examined. To examine whether the susceptibility to TBT-induced apoptosis may be due to the level of glutathione (GSH), we measured the basal GSH levels in Treg and Th2 clones. Intracellular GSH level was measured using high-performance liquid chromatography (HPLC) with a gold electrode. RESULTS: We show that TBT preferentially induces apoptosis in Treg clones rather than in Th2 clones. The basal levels of GSH in Treg clones were significantly lower than those in Th2 clones. CONCLUSIONS: The increased susceptibility of Treg clones to TBT-induced apoptosis appeared to result from lower GSH levels in Treg clones, which may detoxify the reactive oxygen species (ROS) induced by TBT treatment. Our results suggest that the preferential induction of apoptosis in Treg over Th2 contributes to the exacerbation of Th2-driven allergic diseases by TBT.