Male Rat Offspring Research Articles

Long-term effects of cytostatic agents on germ cells of testicular tissue (experimental study)

Currently, cytostatic drugs are widely used not only in cancer treatment, but also in the treatment of autoimmune infammatory diseases. A favorable prognosis of the disease, ability to reproduce, young age and the absence of children serve as an incentive to decide on the need for childbearing. There is concern, that the mutagenic effects of chemotherapy in germ cells, the ability to induce epigenetic changes in them, may have phenotypic manifestations in offspring. Conception in the early stages after treatment (impact on mature and differentiating germ cells) has been proven to increase the risk of defective offspring. Data on the health of the offspring of patients conceived in the long term after treatment (impact on stem spermatogenic cells) are contradictory. The aim of the study was to assess long-term toxic effects of cytostatic drugs in the male rat offspring copulated in terms corresponding to the effect on stem spermatogonial cells (SSCs). Material and Methods. The experiments were carried out on autobred male Wistar rats (n=140), aged 2.5 months, 70 of which made up the group of intact animals. The effect of cytostatic drugs (etoposide, irinotecan, cisplatin, carboplatin, methotrexate, farmorubicin, and paclitaxel) injected 3 and 6 months before mating was assessed on the offspring of intact female and male rats. Results. The male rat offspring treated with cytostatic drugs was found to be viable. Gross external developmental anomalies were detected in 2 cases. In several offspring, a slowdown in physical development, decrease in the rate of formation of sensory-motor refexes and learning ability were observed. The most toxic drugs were etoposide and paclitaxel. Conclusion. The offspring of rats treated with cytostatic drugs in terms corresponding to the effect on the SSCs is at risk. The degree of severity of long-term effects varies signifcantly and depends on the type of the drugs used. A decrease in the ability to learn is the most frequently detected abnormalities in offspring. Judging by the timing of conception after cytostatic exposure, a signifcant increase in the period of time after the administration of the drug before mating is not always justifed.

Gestational exposure to bisphenol S induces microvesicular steatosis in male rat offspring by modulating metaflammation

Prenatal exposure to endocrine-disrupting bisphenol A (BPA) shows a long-lasting programming effect on an organ's metabolic function and predisposes it to the risk of adult metabolic diseases. Although a reduced contaminant risk due to “BPA-free” exposure is proposed, limited data on a comparative assessment of gestational exposure to BPS and BPA and their effects on metaflammation in predisposing liver metabolic disease is reported. Pregnant Wistar rats were exposed to BPS and BPA (0.0, 0.4, 4.0 μg/kg bw) via gavage from gestational day 4 to 21, and effects were assessed in the 90 d male offspring. Prenatal BPS-exposed offspring showed a more obesogenic effect than BPA, including changes in body fat distribution, feed efficiency, and leptin signalling. The BPS exposure induced the adipocyte hypertrophy of visceral adipose to a greater extent than BPA. The adipose hypertrophy was augmented by tissue inflammation, endoplasmic reticulum (ER) stress, and apoptosis due to increased expression of pro-inflammatory (IL6, IL1β, CRP, COX2) cytokines, ER stress modulator (CHOP), and apoptotic effector (Caspase 3). The enlarged, stressed, inflamed adipocytes triggered de novo lipogenesis in the bisphenol-exposed offspring liver due to increased expression of cholesterol and lipid biogenesis mediators (srebf1, fasn, acaca, PPARα) concomitant with elevated triacylglycerol (TG) and cholesterol (TC), resulted in impaired hepatic clearance of lipids. The lipogenic effects were also promoted by increased expression of HSD11β1. BPS exposure increased absolute liver weight, discoloration, altered liver lobes more than in BPA. Liver histology showed numerous lipid droplets, and hepatocyte ballooning, upregulated ADRP expression, an increased expression of pro-inflammatory mediators (IL6, CRP, IL1β, TNFα, COX2), enhanced lipid peroxidation in the BPS-exposed offspring's liver suggest altered metaflammation leads to microvesicular steatosis. Overall, gestational BPS exposure demonstrated a higher disruption in metabolic changes than BPA, involving excess adiposity, liver fat, inflammation, and predisposition to steatosis in the adult male offspring.

Abstract P202: Changes In Cerebral Mitochondrial Function In Intrauterine Growth-Restricted Offspring Before The Development Of Hypertension And Cerebrovascular Dysfunction

Background: Preeclampsia (PE), new-onset gestational hypertension (HTN) affects 3-8% of all births in the USA. PE is the leading cause for intrauterine growth restriction (IUGR). IUGR children have increased risks of HTN and cerebrovascular disease (CVD) later in life. Prior studies in our lab show adult male IUGR rat offspring (OS) to develop HTN, CVD, and cerebral mitochondrial dysfunction (mtDys) at 17 weeks while female IUGR OS exhibit CVD. The role of mtDys in HTN is unknown and is the focus of our study. We hypothesize that IUGR OS will have mtDys prior to HTN. Methods: Our surgical PE animal model utilized the reduced-uterine-perfusion pressure (RUPP) surgery at GD 14. OS from NP served as controls (CON) while OS from RUPP mothers were IUGR. At 12 weeks, OS BP was measured alongside cerebral mtDys through mitochondrial fusion and fission, electron-transport chain (ETC) proteins, and respiration. Results: BP did not change between sexes or among IUGR and CON offspring. Male IUGR OS showed elevated mtRes (State 2: 199 ± 1.08 v. 76 ± 1.13 pmol/mg, p < .05; State 3: 555 ± 3.02 v. 245 ± 1.77 pmol/mg, p < .05; State 4: 346 ± 1.66 v. 85 ± 1.12 pmol/mg, p < .05). Female IUGR and CON OS had no changes in BP or mtRes. Male IUGR OS had increased mito-fusion proteins MFN-1 (176 ± 1.68 v. 100 ± .57 IU/protein/CON %, p < .05), MFN-2 (155 ± 1.36 v. 100 ± .74 IU/protein/CON %, p < .05) and OPA-1 (166 ± 1.99 v. 100 ± .73 IU/protein/CON %, p < .05). DRP-1 had no changes in males; females had no changes in fusion or fission. Male IUGR OS had changes in ETC proteins (CI: 8 ± 9.34 v. 100 ± .82 IU/protein/CON %, p < .05; CII: 23 ± .53 v. 100 ± .52 IU/protein/CON %, p < .05; CIII: 26 ± .23 v. 100 ± .41 IU/protein/CON %, p < .05), as did female IUGR OS (CI: 12 ± .24 v. 100 ± .86 IU/protein/CON %, p < .05; CII: 29 ± .39 v. 100 ± .61 IU/protein/CON %, p < .05; CIII: 39 ± .56 v. 100 ± .62 IU/protein/CON %, p < .05; CIV: 76 ± .74 v. 100 ± .73 IU/protein/CON %, p < .05) vs. CON OS. Summary: Male IUGR OS have elevated mtRes, changed ETC proteins, and fusion proteins that may lead to mtDys and HTN development. This study is clinically relevant because specific cerebral therapy for mtDys for IUGR OS may prevent HTN and CVD development. Future studies will examine changes in mtDys causing male HTN and protection from HTN in females.

DNA fragmentation as a bioindicator of peat fires’ smoke exposure

Introduction. The impact of combustion products of organic substances on the body is known to be accompanied by the accumulation of DNA damage, which can lead to mutations and pathological changes in the cell and the whole organism. The prevalence and scale of this phenomenon poses an important task for studying the consequences that occur in smoke-exposed organisms and their offspring. 
 Materials and methods. The conditions of real peat smoke were reproduced for 40 minutes with a CO concentration of 99±2.5 mg/m3 with using of experimental bio modelling. The genotoxicity of peat fire smoke after exposure to male white rats was assessed by the occurrence of DNA damage in blood cells using the DNA comet method in the alkaline version. In the first part of the experiment, males were directly exposed to the smoke of a peat fire; in the second part – their sexually mature offspring of both sexes were examined for the occurrence of DNA damage in blood cells.
 Results. The males of the parental generation and offspring were found to be resistant to the effects of smoke components, which was confirmed by the absence of statistical significance in terms of «% DNA in the comet tail» compared with the control. At the same time, females of the received generation showed a statistically significant increase in blood cell DNA damage compared to the control group. 
 Limitations. The study was limited to the study of DNA fragmentation after a single 40-minute exposure to peat smoke in male white rats and their intact offspring. 
 Conclusion. The data obtained in this investigation indicate that damage to the DNA structure in the offspring of male rats exposed to peat smoke containing CO at a concentration of 99±2.5 mg/m3 can be considered as a bioindicator of genotoxic effects induced in the next generation.

Saffron and crocin ameliorate prenatal valproic acid-induced autistic-like behaviors and brain oxidative stress in the male offspring rats.

Autism is a neurobehavioral disease that induces cognitive and behavioral alterations, usually accompanied by oxidative stress in the brain. Crocus sativus (saffron) and its active ingredient, crocin, have potent antioxidative effects that may benefit autistic behaviors. This study aimed to determine the effects of saffron extract and crocin against brain oxidative stress and behavioral, motor, and cognitive deficits in an animal model of autism in male offspring rats. 14 female rats were randomly divided into the saline and valproic acid (VPA) groups. Then, they were placed with mature male rats to mate and produce offspring. VPA (500mg/kg, i.p.) was injected on day 12.5 of pregnancy (gestational day, GD 12.5) to induce an experimental model of autism. 48 male pups were left undisturbed for 29days. First-round behavioral tests (before treatments) were performed on 30-33 post-natal days (PND), followed by 28days of treatment (PND 34-61) with saffron (30mg/kg, IP), crocin (15 or 30mg/kg, i.p.), or saline (2ml/kg, i.p.). The second round of behavioral tests (after treatments) was performed on PND 62-65 to assess the effects of the treatments on behavioral and cognitive features. In the end, animals were sacrificed under deep anesthesia, and their brains were dissected to evaluate the brain oxidative stress parameters, including malondialdehyde (MDA), glutathione (GSH), and catalase (CAT). VPA injection into female rats increased anxiety-like behaviors, enhanced pain threshold, impaired motor functions, disturbed balance power, increased MDA, and decreased GSH and CAT in their male offspring. 28days of treatment with saffron or crocin significantly ameliorated behavioral abnormalities, reduced MDA, and increased GSH and CAT levels. Brain oxidative stress has been implicated in the pathophysiology of autistic-like behaviors. Saffron and crocin ameliorate anxiety-like behaviors, pain responses, motor functions, and brain oxidative stress parameters in an experimental model of autism. Saffron and crocin may hold promise as herbal-based pharmacological treatments for individuals with autism. However, further histological evidence is needed to confirm their efficacy.

Assessment of Ante- and Postnatal Development of the Offspring of Male Rats Crossed in Delayed Periods after Treatment with Methotrexate in Low Doses.

We studied ante- and postnatal development of the offspring of intact female rats crossed with males injected with low doses of methotrexate 3 and 6 months before mating. The time of crossing corresponded to the manifestation of the cytostatic effect on spermatogonial stem cells. The offspring of methotrexate-treated males was characterized by increased preimplantation losses and fetal growth restriction in the antenatal period and inhibition of physical development, delayed formation of sensory-motor reflexes, and impaired learning abilities in the postnatal period.

Melatonin Corrects Depressive-like Behaviour and Anxiety in Male and Female Offspring Rats in a Model of Prenatal Melatonin Deficit

Melatonin deficiency caused by a prolonged light regime leads to a disturbance in the circadian rhythmicity of a number of physiological and biochemical parameters, including disruption of the synchronizing role of the hormone on endocrine and metabolic functions in the body. The aim of the present study was to explore the sex-dependent consequences of prenatal exposure to constant light (CL) on behavioural responses of rat adult offspring. In addition, the effect of melatonin supplementation on pregnant rats exposed to CL was evaluated on emotional disturbance due to melatonin deficiency. Chronic exposure to CL during pregnancy induced impaired emotional responses in male and female adult offspring, including decreased visits to aversive central zone in the open field test and elevated plus maze test and reduced consumption of sweet solutions. The elevated anxiety and anhedonia was corrected in a generation of prenatal melatonin-deficient rats of the two sexes. Our results suggest that prenatal rat exposure to CL provokes long-term behavioural impairments in male and female adult offspring. The beneficial role of melatonin supplementation on these disturbed responses suggests that our prenatal model induces melatonin deficiency that causes sustainable changes in offspring. Further data are needed to ascertain the mechanism underlying the prenatal-induced deleterious effect of melatonin deficiency and possible therapeutic options.

Protective role of vitamin E against acrylamide-induced testicular toxicity from pregnancy to adulthood: insights into oxidative stress and aromatase regulation.

Acrylamide (ACR) is a toxic chemical frequently encountered in daily life, posing health risks. This study aimed to elucidate the molecular-level mechanism of ACR's toxic effects on testicles and investigate whether Vitamin E can mitigate these effects. A total of 40 adult pregnant rats were utilized, divided into four groups: Control, ACR, Vitamin E, and ACR + Vitamin E. ACR and Vitamin E were administered to the mother rats during pregnancy and lactation, and to the male offspring until the 8th week post-birth. Serum hormone levels, oxidant-antioxidant parameters, histopathological examination of testicular tissue, and mRNA and protein levels of the testicular and liver aromatase gene were analyzed. Spermiogram analysis was conducted on the collected sperm samples from the male offspring. The results revealed that ACR exposure adversely affected hormone levels, oxidant-antioxidant parameters, histological findings, as well as aromatase gene and protein expressions. However, Vitamin E administration effectively prevented the toxic effects of ACR. These findings demonstrate that ACR application significantly impairs the reproductive performance of male offspring rats by increasing liver aromatase activity.

Dietary Zinc Status in Offspring of Pregnant Rats Fed on a Zinc-Deficient Diet Is Associated with Serum Albumin, Ast, and Alt Levels

Objective: This study was carried out to investigate whether dietary zinc status is associated with serum albumin, AST and ALT levels in male offspring of mother rats fed a zinc deficient diet. Methods: The study was carried out on male offspring (Groups 1, 2, 3) born to rats fed a zinc deficient diet and on male offspring (Group 4) born to mothers fed a standard diet. Group 1: Zinc deficient, Group 2; standard rat chow, Group 3: Zinc supplemented diet. Animals of group 4 were used as control group. After the completion of the experimental stages of the study, albumin, AST, ALT, free and total bilirubin levels in serum samples taken from animals were determined by spectrophotometric method. Results: Dietary zinc deficiency (group 1) significantly decreased serum albumin values (p<0.004). Animals in both the zinc deficient (Group 1) group and the Group 2 animals born to mothers fed a zinc deficient diet and fed standard rat chow had the highest AST and ALT levels (p<0.001). Conclusion: The present study is the first to show that dietary zinc status can directly affect liver function in rats born to zinc deficient mothers by causing changes in serum albumin, AST and ALT levels.

Islet hypoplasia of adult offspring rats caused by intrauterine chronic hypoxia is compensated by up-regulation of INS and PDX-1

ABSTRACT Background: Intrauterine chronic hypoxia (ICH) can lead to pancreatic dysmetabolism in offspring. This study aimed to determine the changes in islet function of offspring through a rat ICH model and detect the factors affecting islet function. Methods: Twenty couples of healthy Sprague – Dawley adult rats were randomly mated, and the pregnant rats were randomly allocated to ICH and normal control (NC) groups. Pregnant rats in the ICH group were placed in a hypoxic chamber with 13% oxygen concentration for hypoxia treatment twice a day for 4 h until delivery at 21 days. NC group is inlet with normal air from beginning to end. After delivery, blood was taken from the heart of pregnant rats for blood gas analysis. The weight of the offspring rats was measured at 12 h after birth and 16 weeks after birth. At 16 weeks, the immunohistochemical results of β-cell total, islet area, insulin (INS), and glucose transporter 2 (GLUT2) proteins were obtained from the islets. The mRNA data of INS and pancreatic and duodenal homeobox 1 (PDX-1) genes were obtained from pancreas. Results: We found the β-cell total, islet area, and the positive cell area of INS and GLUT2 of offspring rats in ICH group were lower than those of NC group, while the levels of INS and PDX-1 genes were higher in ICH group than in NC group. Conclusions: ICH can lead to islet hypoplasia in adult male offspring rats. However, this is within the compensatory range.

Transgenerational Effects and Mechanisms of Tributyltin Exposure on Neurodevelopment in the Male Offspring of Rats.

This study aimed to investigate the transgenerational effects of tributyltin exposure on rat neurodevelopment in male offspring and the potential mechanisms. Neonatal female rats were exposed to the environmental level of tributyltin and then mated with nonexposed males after sexual maturity to produce the F1 generation. The F1 generation (with primordial germ cell exposure) was mated with nonexposed males to produce nonexposed offspring (the F2 and F3 generations). Neurodevelopmental indicators and behavior were observed for the F1, F2, and F3 generations during postnatal days 1-25 and 35-56, respectively. We found premature eye-opening and delayed visual positioning in newborn F1 rats and anxiety and cognitive deficits in prepubertal F1 male rats. These neurodevelopmental impacts were also observed in F2 and F3 males. Additionally, F1-F3 males exhibited increased serotonin and dopamine levels and a loose arrangement of neurons in the hippocampus. We also observed a reduction in the expression of genes involved in intercellular adhesion and increased DNA methylation of the Dsc3 promoter in F1-F3 males. We concluded that tributyltin exposure led to transgenerational effects on neurodevelopment via epigenetic reprogramming in male offspring. These findings provide insights into the risks of neurodevelopmental disorders in offspring from parents exposed to tributyltin.

Effects of prenatal nicotine exposure on vascular endothelial function and structure in offspring rats

To clarify the effect of prenatal nicotine exposure (PNE) on vascular endothelial function (VEF) in offspring rats, and to explore whether these effects are long-lasting and sex-dependent. Pregnant Sprague-Dawley rats were randomized into two groups and exposed to either 102 mg/mL nicotine (experimental group) or normal saline (control group) through a subcutaneously implanted osmotic micropump. Vascular rings from the thoracic aorta were collected from offspring rats at 1, 4, and 12 months of age. The specimens were used to determine VEF using functional tests and to observe morphologic and pathologic changes of blood vessels in hematoxylin and eosin-stained samples with light microscopy. The percentage of acetylcholine-induced endothelium-dependent vasorelaxation (EDV) of the thoracic aorta was significantly higher in the experimental group than in the control group in 1-, 4-, and 12-month-old female offspring rats (18.68±2.32 vs. 13.85±6.10, 17.44±3.91 vs. 10.77±5.77, and 30.96±22.26 vs. 18.94±13.61, respectively; P <0.05). The percentage of acetylcholine-induced EDV was significantly lower in the experimental group than in the control group in 1-, 4-, and 12-month-old male offspring rats (10.31±4.05 vs. 16.05±14.80, 5.57±2.81 vs. 12.12±5.62, and 11.98±7.24 vs. 58.87±32.43, respectively; P <0.05). The hematoxylin & eosin-stained vascular rings of the offspring rats displayed an uneven lumen, scattered intimal thickening, partial shedding and vacuolar degeneration of endothelial cells, and inflammatory cell infiltration and phagocytosis in the experimental group. In addition, overt smooth muscle atrophy in tunica media, disordered cell arrangement, and unclear structure of the elastic fiber layer was observed. None of these histopathologic changes were found in the control group. In adult offspring rats, PNE not only affected the VEF of the thoracic aorta but also led to pathologic changes in the vascular structure. Additionally, the effects of PNE on VEF were sex-specific, manifesting primarily as significantly improved VEF in female offspring rats and significantly impaired VEF in male offspring rats, lasting into adulthood.

Maternal High-Fat Diet Multigenerationally Programs HPA Function and Behaviors in Male Rat Offspring.

Maternal environmental factors have been demonstrated to exert significant influences on the health of offspring. The hypothalamic-pituitary-adrenal (HPA) axis is an important neuroendocrine stress system that can be influenced by early life challenges. Our previous research has revealed that the consumption of a high-fat diet (HFD) by pregnant and lactating rats leads to the programming of HPA axis activity in male offspring of the first generation (referred to as F1HFD/C). The present study aimed to investigate whether the observed remodeling of the HPA axis could be inherited by second-generation male offspring (referred to as F2HFD/C), following maternal HFD exposure. The results showed that F2HFD/C rats exhibited enhanced basal HPA axis activity, similar to their F1HFD/C ancestors. Moreover, F2HFD/C rats displayed exacerbated corticosterone responses to restraint and lipopolysaccharide-induced stress, but not to insulin-induced hypoglycemia stress. Furthermore, maternal HFD exposure significantly aggravated depression-like behavior in the F2 generation subjected to chronic unpredictable mild stress. To investigate the role of central calcitonin gene-related peptide (CGRP) signaling in maternal diet-induced programming of the HPA axis across generations, we conducted central infusion of αCGRP8-37, a CGRP receptor antagonist, in F2HFD/C rats. The results demonstrated that αCGRP8-37 attenuated depression-like behaviors and reduced the hyperresponsiveness of the HPA axis to restraint stress in these rats. Therefore, central CGRP signaling may contribute to maternal diet-induced programming of HPA axis across generations. In conclusion, our study provides evidence that maternal HFD consumption can lead to multigenerational programming of the HPA axis and behaviors in adult male descendants.

Impact of Maternal Exposure to Lead Acetate before Pregnancy Through Lactation Period on the Testicular histomorphologic Indices of Male Offspring of Wistar Rats, A Stereological Study

Impact of Maternal Exposure to Lead Acetate before Pregnancy Through Lactation Period on the Testicular histomorphologic Indices of Male Offspring of Wistar Rats, A Stereological Study

Developmental deficits in male rat pups caused by maternal and dietary administration of chlorpyriphos and cypermethrin: Melatonin's mitigating effect.

The ability of melatonin to reduce growth inadequacy induced by parental and nutritional combination administration to chlorpyriphos (Ch) and cypermethrin (Cy) was examined in male albino rats. Oral alimentation was given to gravid dams divided into six groups (n = 10; age: 12weeks) from the first day of pregnancy to the 21st postnatal day. Distilled water (DW), Soya oil (SYO), and melatonin (MeL) groups were exposed to 2mL/kg, 2mL/kg, and 0.5mg/kg, respectively; the Ch+Cy group was co-exposed to Ch (1.9mg/kg of LD50) and Cy (7.5mg/kg of LD50); the MChCy group was preconditioned with MeL (0.5mg/kg), followed by co-exposure to Ch and Cy; and the ChCyM group was exposed to Ch and Cy and post treated with MeL. Male offspring rats were tested for ontogeny criteria at various points after accouchement. MeL pre- and post-administration reduced the variation in litter size and weight, number of live/dead pups, anogenital distance , crown-rump length, the timing of eye and ear openings, and testicular descent caused by fetal and nutritional co-administration to Ch+Cy in offspring male albino rats. MeL demonstrated preventive promise as a result of its apparent antioxidative capability.

Autophagy inhibition mediated by intrauterine miR-1912-3p/CTSD programming participated in the susceptibility to osteoarthritis induced by prenatal dexamethasone exposure in male adult offspring rats.

Autophagy inhibition is known to be involved in the development of adult osteoarthritis. Dexamethasone, as a synthetic glucocorticoid, is widely used for premature delivery and related pregnancy diseases in clinics. We have previously shown that prenatal dexamethasone exposure (PDE) was associated with increased susceptibility to postnatal osteoarthritis in offspring. However, whether the occurrence of fetal-originated adult osteoarthritis induced by PDE is related to autophagy remains unclear. In this study, we first found that PDE could increase the mRNA and protein expression of cartilage matrix-degrading enzymes (MMP3, MMP13, and ADAMTS5) and decrease the cartilage matrix contents in adult offspring, and the in vitro results suggested that this might be related to the autophagy inhibition of chondrocytes. Further, we demonstrated a persistent autophagy inhibition with autolysosome accumulation, low expression of cathepsin D (CTSD), increased H3K9ac level, and expression of miR-1912-3p in the cartilage of PDE offspring from fetus to adulthood. In vitro experiments showed that dexamethasone inhibited autophagy flux and CTSD expression in fetal chondrocytes, while overexpression of CTSD could alleviate the inhibition of autophagic flux induced by dexamethasone. Finally, we confirmed that dexamethasone increased the H3K9ac level and expression of miR-1912-3p through activation of the glucocorticoid receptor (GR), resulting in the decreased expression of CTSD and inhibition of autophagy flux in fetal chondrocytes. In conclusion, intrauterine miR-1912-3p/CTSD programming-mediated autophagy inhibition promoted the susceptibility to osteoarthritis in PDE adult offspring rats. This study provides new ideas for exploring early prevention and therapeutic targets in fetal-originated osteoarthritis.

Urinary microbiota and metabolic signatures associated with inorganic arsenic-induced early bladder lesions

Inorganic arsenic (iAs) contamination in drinking water is a global public health problem, and exposure to iAs is a known risk factor for bladder cancer. Perturbation of urinary microbiome and metabolome induced by iAs exposure may have a more direct effect on the development of bladder cancer. The aim of this study was to determine the impact of iAs exposure on urinary microbiome and metabolome, and to identify microbiota and metabolic signatures that are associated with iAs-induced bladder lesions. We evaluated and quantified the pathological changes of bladder, and performed 16S rDNA sequencing and mass spectrometry-based metabolomics profiling on urine samples from rats exposed to low (30 mg/L NaAsO2) or high (100 mg/L NaAsO2) iAs from early life (in utero and childhood) to puberty. Our results showed that iAs induced pathological bladder lesions, and more severe effects were noticed in the high-iAs group and male rats. Furthermore, six and seven featured urinary bacteria genera were identified in female and male offspring rats, respectively. Several characteristic urinary metabolites, including Menadione, Pilocarpine, N-Acetylornithine, Prostaglandin B1, Deoxyinosine, Biopterin, and 1-Methyluric acid, were identified significantly higher in the high-iAs groups. In addition, the correlation analysis demonstrated that the differential bacteria genera were highly correlated with the featured urinary metabolites. Collectively, these results suggest that exposure to iAs in early life not only causes bladder lesions, but also perturbs urinary microbiome composition and associated metabolic profiles, which shows a strong correlation. Those differential urinary genera and metabolites may contribute to bladder lesions, suggesting a potential for development of urinary biomarkers for iAs-induced bladder cancer.

EARLY PPARα ACTIVATION PROTECT AGAINST HEPATIC MICROSTEATOSIS INDUCED BY POST-NATAL OVERFEEDING

Post-natal overfeeding leads to development of obesity and metabolic dysfunction. PPARαactivation improves steatosis and metabolic dysfunction associated with obesity. Thus, wehave hypothesized that PPARα would prevent the metabolic disorders and obesity caused bylitter reduction. Wistar male offspring rats were divided into normal litter (NL - 9 pups) andsmall litter (SL - 3 pups) and were daily treated with an PPARα agonist, fenofibrate, frompost-natal day (PN) 1 until weaning (PN21), with either, fenofibrate (F - 12.5mg/kg) orvehicle (V). Forming the following groups: NL-V, NL-F, SL-V, SL-F. At PN120 animalswere euthanized and histological evaluation of samples from liver, visceral (VAT),subcutaneous (SAT) and brown (BAT) adipose tissue were performed. Fenofibrate was ableto prevent the development of microsteatosis in the liver caused by postnatal overfeeding. Inregard to VAT, there was no difference in the mean adipocyte area, however in distributionanalyzes it was observed higher frequency of larger adipocytes in SL groups, and nodifference was observed in SAT. SL-V animals had increased lipid area in BAT, which wasreduced in SL-F. Increased PPARα activation induced by an agonist during lactationimproved some histopathological abnormalities induced by postnatal overfeeding. This wasproven by the decrease in microsteatosis in the liver and improvement in VAT and BATmorphology, thus implying a better metabolic health.

MATERNAL FOOD RESTRICTION DURING LACTATION LEADS TO SEX-SPECIFIC BLOOD HYPERTENSION AND CARDIAC REMODELING IN THE OFFSPRING

Dietary restriction during pregnancy and puberty is a risk factor to hypertension onset atadulthood. The aim of this study was to evaluate the effect of maternal malnutrition duringlactation on cardiovascular parameters, in both sexes of rat pups at 120-day-old. Maternalfood restriction lead to arterial hypertension, baroreflex impairment, cardiac remodeling andincreased collagen deposition in the aorta, in adult male offspring. The females did not showfunctional or morphological disruptions. Together, these results show that maternal foodrestriction during lactation promotes cardiometabolic programming, generating cardiovascularalterations in male adult rat offspring, however female offspring present some kind ofcardioprotective effect.

MATERNAL PROTEIN MALNUTRITION DURING LACTATION PREDISPOSES TO A METABOLIC SYNDROME PHENOTYPE IN MALE WISTAR RATS

Maternal nutritional insults during lactation can modulate the offspring phenotype associatedwith the risk of non-communicable diseases at different stages of ontogenetic development.The aim of this research was to analyze biometric and biochemical parameters of male Wistarrats, born to dams fed a low-protein diet in the first two-thirds of lactation. Therefore, themothers received ad libitum a normal (NP, 23%) or low-protein (LP, 4%) diet, originatingtwo experimental groups that were evaluated at 14 days old (NP-14 and LP-14). The resultsshowed that the LP-14 offspring male rats had lower body weight (p<0.0001), reduced liver(p<0.0001) and higher brown fat deposits (p<0.0001), compared to the equivalent control.They also exhibited hyperglycemia (p<0.05), hypercholesterolemia (p<0.0001), and increasedserum β-hydroxybutyrate (p<0.0001), contrasting with reduced triglycerides (p<0.05) levels.We conclude that maternal exposure to a low-protein diet during lactation was able toprogram 14-day-old male neonates to develop a metabolic syndrome phenotype, probably dueto restriction of body and visceral growth in these animals.