Wistar Rat Pups Research Articles

Neuroprotective Effect of Melatonin in a Neonatal Hypoxia-Ischemia Rat Model Is Regulated by the AMPK/mTOR Pathway.

Melatonin has been shown to be neuroprotective in different animal models of neonatal hypoxic-ischemic brain injury. However, its exact molecular mechanism of action remains unknown. Our aim was to prove melatonin's short- and long-term neuroprotection and investigate its role on the AMPK (AMP-activated protein kinase)/mTOR (mammalian target of rapamycin) pathway following neonatal hypoxic-ischemic brain injury. Seven-day-old Wistar rat pups were exposed to hypoxia-ischemia, followed by melatonin or vehicle treatment. Detailed analysis of the AMPK/mTOR/autophagy pathway, short- and long-term neuroprotection, myelination, and oligodendrogenesis was performed at different time points. At 7 days after hypoxia-ischemia, melatonin-treated animals showed a significant decrease in tissue loss, increased oligodendrogenesis, and myelination. Long-term neurobehavioral results showed significant motor improvement following melatonin treatment. Molecular pathway analysis showed a decrease in the AMPK expression, with a significant increase at mTOR's downstream substrates, and a significant decrease at the autophagy marker levels in the melatonin group compared with the vehicle group. Melatonin treatment reduced brain area loss and promoted oligodendrogenesis with a clear improvement of motor function. We found that melatonin associated neuroprotection is regulated via the AMPK/mTOR/autophagy pathway. Considering the beneficial effects of melatonin and the results of our study, melatonin seems to be an optimal candidate for the treatment of newborns with hypoxic-ischemic brain injury in high- as well as in low- and middle-income countries.

Evaluation of developmental milestones and of brain measurements in rats exposed to the pesticide pyriproxyfen in prenatal period.

Pyriproxyfen is a pesticide used in Brazil to control the Aedes aegypti mosquito, vector of arboviruses like Zika and dengue. However, this pesticide is structurally similar to retinoic acid, a metabolite of vitamin A that regulates neuronal differentiation and hindbrain development during the embryonic period. Due to the similarity between pyriproxyfen and retinoic acid, studies indicate that this pesticide may have cross-reactivity with retinoid receptors. Thus, pregnant exposure to pyriproxyfen could interfere in the nervous system development of the fetal. In this context, the present study evaluated whether prenatal exposure to pyriproxyfen affects neonatal development and brain structure in rats. Wistar rat pups were divided in three experimental groups: (1) negative control (CT-)-offspring of rats that drink potable water during pregnancy; (2) pyriproxyfen (PIR)-offspring of rats exposed to Sumilarv® prenatally, a pesticide that has pyriproxyfen as active ingredient; and (3) positive control (CT+)-offspring of rats exposed to an excess of vitamin A prenatally. Only vitamin A treated-pregnant showed lower weight gain, but gestation length was similar among pregnant that received potable water, water containing vitamin A and water containing Sumilarv. In relation to the offspring, PIR group exhibits a delayed front-limb suspension response but performed early the negative geotaxis reflex. On the other hand, CT+ group exhibited lower body weight in the 1st postnatal day, delayed audio startle response, but performed early the eyelids opening and hindlimb placing response. A reduction in the maximum brain width was observed both in PIR and CT+ groups, but a reduction in the number of neurons in the M1 cortex was showed only in CT+ group. The number of glial cells in this brain area was similar between the three experimental groups studied. Although prenatal exposure to pyriproxyfen did not alter neonatal milestones in the same way as vitamin A in excess, both substances caused a reduction in the maximum width of the brain, suggesting that this pesticide can produce neurotoxic effects during the embryonic period.

Cerium oxide nanoparticles (nanoceria) pretreatment attenuates cell death in the hippocampus and cognitive dysfunction due to repeated isoflurane anesthesia in newborn rats

General anesthetics exposure, particularly prolonged or repeated exposure, is a crucial cause of neurological injuries. Notably, isoflurane (ISO), used in pediatric anesthesia practice, is toxic to the developing brain. The relatively weak antioxidant system at early ages needs antioxidant support to protect the brain against anesthesia. Cerium oxide nanoparticles (CeO2-NPs, nanoceria) are nano-antioxidants and stand out due to their unique surface chemistry, high stability, and biocompatibility. Although CeO2-NPs have been shown to exhibit neuroprotective and cognitive function-facilitating effects, there are no reports on their protective effects against anesthesia-induced neurotoxicity and cognitive impairments. Herein, Wistar albino rat pups were exposed to ISO (1.5 %, 3-h) at postnatal day (P)7+P9+P11, and the protective properties of CeO2-NP pretreatment (0.5 mg/kg, intraperitoneal route) were investigated for the first time. The control group at P7+9+11 received 50 % O2 (3-h) instead of ISO. Exposure to nanoceria one-hour before ISO protected hippocampal neurons of the developing rat brain against apoptosis [determined by hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) analysis with caspase-3, and immunoblotting with Bax/Bcl2, cleaved caspase-3 and PARP1] oxidative stress, and inflammation [determined by immunoblotting with 4-hydroxynonenal (4HNE), nuclear factor kappa-B (NF-κB), and tumor necrosis factor-alpha (TNF-α)]. CeO2-NP pretreatment also reduced ISO-induced learning (at P28–32) and memory (at P33) deficits evaluated by Morris Water Maze. However, memory deficits and thigmotactic behaviors were detected in the agent-control group; elimination of these harmful effects will be possible with dose studies, thus providing evidence supporting safer use. Overall, our findings support pretreatment with nanoceria application as a simple strategy that might be used for pediatric anesthesia practice to protect infants and children from ISO-induced cell death and learning and memory deficits.

Effects of Memantine on Cognitive Flexibility in a Valproic Acid Rat Model of Autism

Abstract Autistic children frequently struggle with cognitive flexibility. The potential molecular mechanism underlying cognitive inflexibility remains unclear. Decreased level of brain-derived neurotrophic factor (BDNF), which plays a vital role in neural development and synaptic plasticity may have a role. This study aimed to investigate the effects of memantine on (BDNF) expression in hippocampus of prenatal VPA exposed rats, and its consequent effects on cognitive flexibility. Pregnant female rats received a single intraperitoneal injection of VPA (600 mg/kg) on gestational day 12.5 to induce autistic-like behaviors in their offspring. Afterwards, the 23-day-old male Wistar rat pups received daily intraperitoneal injections of memantine (20 mg/kg) for 4 weeks. The prenatal VPA-exposed rats exhibited reduced cognitive flexibility in the attentional set- shifting task (ASST) reflecting ASD-associated cognitive flexibility deficits, a common ASD- associated problem with accompanied reduction in hippocampal BDNF. The defective performance in ASST was more prominent in the reversal stages. Memantine associated increased hippocampal BDNF could alleviate cognitive inflexibility in ASD. This study draws attention to the potential favorable effect of memantine on cognitive performance in autistic subjects.

Melanocortin-receptor 4 activation modulates proliferation and differentiation of rat postnatal hippocampal neural precursor cells

Postnatal hippocampal neurogenesis is essential for learning and memory. Hippocampal neural precursor cells (NPCs) can be induced to proliferate and differentiate into either glial cells or dentate granule cells. Notably, hippocampal neurogenesis decreases dramatically with age, partly due to a reduction in the NPC pool and a decrease in their proliferative activity. Alpha-melanocyte-stimulating hormone (α-MSH) improves learning, memory, neuronal survival and plasticity. Here, we used postnatally-isolated hippocampal NPCs from Wistar rat pups (male and female combined) to determine the role of the melanocortin analog [Nle4, D-Phe7]-α-MSH (NDP-MSH) in proliferation and fate acquisition of NPCs. Incubation of growth-factor deprived NPCs with 10 nM NDP-MSH for 6 days increased the proportion of Ki-67- and 5-bromo-2′-deoxyuridine (BrdU)-positive cells, compared to the control group, and these effects were blocked by the MC4R antagonist JKC-363. NDP-MSH also increased the proportion of glial fibrillar acidic protein (GFAP)/Ki-67, GFAP/sex-determining region Y-box2 (SOX2) and neuroepithelial stem cell protein (NESTIN)/Ki-67-double positive cells (type-1 and type-2 precursors). Finally, NDP-MSH induced peroxisome proliferator-activated receptor (PPAR)-γ protein expression, and co-incubation with the PPAR-γ inhibitor GW9662 prevented the effect of NDP-MSH on NPC proliferation and differentiation. Our results indicate that in vitro activation of MC4R in growth-factor-deprived postnatal hippocampal NPCs induces proliferation and promotes the relative expansion of the type-1 and type-2 NPC pool through a PPAR-γ-dependent mechanism. These results shed new light on the mechanisms underlying the beneficial effects of melanocortins in hippocampal plasticity and provide evidence linking the MC4R and PPAR-γ pathways in modulation of hippocampal NPC proliferation and differentiation.

Effects of a Functional Food Made with Salvia hispanica L. (Chia Seed), Amaranthus hypochondriacus L. (Amaranth), and an Ethanolic Extract of Curcuma longa L. (Curcumin) in a Rat Model of Childhood Obesity.

Obesity is a global health problem and is increasing in prevalence in most countries. Although obesity affects all age groups, children are the most vulnerable sector. Functional foods are novel formulated foods containing substances (i.e., nutrients, phytochemicals, probiotics, etc.) that have potential health-enhancing or disease-preventing value. The research objective was to study the possible beneficial effects of providing a functional food made with amaranth flour, chia seed, and curcumin extract on the metabolism and behavior of a rat model of childhood obesity. Male Wistar rat pups from two litters of different sizes, a normal litter (NL) (10 pups) and a small litter (SL) (4 pups), were used. After weaning, the rats were fed a hypercaloric diet (HD) or an HD supplemented with the functional food mixture. Body weight and energy intake were measured for seven weeks, and locomotor activity, learning, and memory tests were also performed. At the end of the experiment, glucose and lipid metabolism parameters were determined. The results showed that in this model of obesity produced by early overfeeding and the consumption of a hypercaloric diet, anxiety-like behaviors and metabolic alterations occurred in the rat offspring; however, the provision of the functional food failed to reduce or prevent these alterations, and an exacerbation was even observed in some metabolic indicators. Interestingly, in the NL rats, the provision of the functional food produced some of the expected improvements in health, such as significant decreases in body weight gain and liver cholesterol and non-significant decreases in adipose tissue and leptin and insulin serum levels.

Crocin Exerts Neuroprotective Effect in Valproate Exposed Rat Model of Autistic Spectrum Disorder by Ameliorating Behavioural and Biochemical Reversions

Autistic spectrum disorder (ASD) is associated with oxidative stress and neuron damage triggered by valproic acid (VPA) in a post-natal rat model of ASD. It was hypothesized that crocin (CRO), may have beneficial effects in ASD due to its antioxidant activities. The study investigated the neuro-behavioural and biochemical modifications of crocin in VPA-induced ASD. Wistar albino rat pups aged 13 days were divided into five groups (06 each). 400 mg/kg VPA subcutaneously on PND 14 successfully induced autism. From PND 14 to 40, crocin was dispensed daily. Different neuro-behavioral markers were examined while the groups were receiving therapy. Animals were sacrificed at the completion of the research for biochemical calculations. Crocin treatment significantly improved behavioral activity and oxidative changes compared to animals treated with the VPA-exposed group. The effects were more pronounced at higher dose of crocin, providing evidence of reversing the valproic acid-induced autistic deficits. Crocin could be a viable option for ASD treatment due to its potential neuronal cytoprotective effects, likely attributed to its antioxidant properties

Sulbactam protects neurons against double neurotoxicity of amyloid beta and glutamate load by upregulating glial glutamate transporter 1

Amyloid beta (Abeta) synergistically enhances excitotoxicity of glutamate load by impairing glutamate transporter 1 (GLT1) expression and function, which exacerbates the development of Alzheimer’s disease (AD). Our previous studies suggested that sulbactam can upregulate the expression levels and capacity of GLT1. Therefore, this study aims to investigate whether sulbactam improves neuronal tolerance against neurotoxicity of Abeta and glutamate load by up-regulating GLT1 in primary neuron-astrocyte co-cultures. Early postnatal P0–P1 Wistar rat pups’ cortices were collected for primary neuron–astrocyte cultures. Hoechst–propidium iodide (HO–PI) stain and lactate dehydrogenase (LDH) assays were used to analyze neuronal death. Cell counting kit 8 (CCK8) was applied to determine cell viability. Immunofluorescence staining and western blotting were used to assess protein expressions including GLT1, B-cell lymphoma 2 (BCL2), BCL2 associated X (BAX), and cleaved caspase 3 (CCP3). Under the double effect of Abeta and glutamate load, more neurons were lost than that induced by Abeta or glutamate alone, shown as decreased cell viability, increased LDH concentration in the cultural medium, HO–PI positive stains, high CCP3 expression, and high BAX/BCL2 ratio resulting from increased BAX and decreased BCL2 expressions. Notably, pre-incubation with sulbactam significantly attenuated the neuronal loss and activation of apoptosis induced by both Abeta and glutamate in a dose-dependent manner. Simultaneously, both astrocytic and neuronal GLT1 expressions were upregulated after sulbactam incubation. Taken together, it could be concluded that sulbactam protected neurons against double neurotoxicity of Abeta and glutamate load by upregulating GLT1 expression. The conclusion provides evidence for potential intervention using sulbactam in AD research.

Oral intake of bovine lactoferrin alleviates intestinal injury induced by perinatal hypoxia and hypothermia in newborn rats

Introduction/Objectives. To investigate whether oral administration of lactoferrin attenuates intestinal injury induced by perinatal hypoxia and hypothermia. Methods. Newborn Wistar rat pups were exposed to perinatal asphyxia, followed by global hypothermia. The pups were distributed into two groups: lactoferrin group (LG) ? pups that received lactoferrin orally for seven days (20 mg/day), control group (CG) ? pups that received normal saline orally during first seven days of life. After seven days macroscopic examination of the bowels and pathohistological analyses of tissue samples have been performed. Results. The incidence of macroscopic injuries was significantly lower in LG group compared to CG. The incidence of pathological findings, as well as the values of injury scores used to assess the intensity and extent of intestinal injury at microscopic level were significantly lower in LG group. Conclusions. Lactoferrin attenuates perinatal hypoxia/hypothermia-induced intestinal injury in newborn rats.

Impact of Mucuna pruriens seed on lipid profiles in rats with streptozotocin-induced diabetes

Background: Mucuna pruriens (MP) is a tropical legume, widely used in the treatment of parkinsonism, erectile dysfunction, anti-venomous, and anti-diabetic properties. It contains multiple phytonutrients and is rich in soluble fibers and proteins and is used as a drug of choice in traditional medicine for different ailments. Aims and Objectives: The study aimed to study the hypolipidemic activity of alcoholic extract MP seeds in Type 2 diabetic rats with hyperlipidemia, by assessing lipid profile of rats in comparison with rats treated with a standard anti-diabetic drug. Materials and Methods: Fifty-four Wistar rat pups were made diabetic by injecting intraperitoneal injection of streptozotocin (65 mg/kg). After 12 weeks, animals with diabetic and dyslipidemia were selected and divided into three groups diabetic control, test drug group (fed with MP alcoholic extract 200 mg/kg), and standard drug group (fed with glibenclamide 0.1 mg/kg), and 18 rats of same age were kept for normal control group. Results: MP seeds exhibited a gradual and progressive change in the low-density lipoprotein levels in diabetic rats (4th week [−34.3%], 8th week [−42.24%], and 12th week [−45.30%]), serum triglycerides levels (4th week [−39.3%], 8th week [−44.49%], and 12th week [−46.65%]). Whereas serum high-density lipoprotein levels were significantly increased (39.3% increase by 4 weeks, 54.95% increase by 8 weeks, and 61.03% increase by 12 weeks). This suggests that active phytochemical compounds in the MP seeds exert hypolipidemic properties by its antioxidant and high fiber content. Conclusion: MP seeds contain phytonutrients and soluble fiber compounds, which helps in reduction of lipid levels in rats with diabetes induced hyperlipidemia.

The role of SIRT3 in mediating the cognitive deficits and neuroinflammatory changes associated with a developmental animal model of schizophrenia

The neuroinflammatory state may contribute to the pathogenesis of many mental disorders including schizophrenia. Nicotinamide adenine dinucleotide (NAD+) is an essential cofactor for activation of proteins involved in mitochondria quality control, such as Sirtuin3 (SIRT3). Our previous study has found that NAD+ supplement could rescue early life stress (ELS)-induced neuroinflammation and down-regulation of SIRT3 in adult offspring. However, it is unclear whether SIRT3 is the key to the neuroprotective effects of NAD+ supplement in this animal model of schizophrenia. The present study used 24 h maternal separation (MS) as ELS to Wistar rat pups on the postnatal day (PND) 9. Schizophrenia-like behaviors and memory impairments were detected by behavioral tests. Microglial activation, pro-inflammatory cytokine expression, and NAD+/SIRT3 expression were detected in the prefrontal cortex and hippocampus. Meanwhile, NAM (a precursor of NAD+), and the SIRT3 activator Honokiol (HNK), and the SIRT3 inhibitor 3-TYP were used as an intervention in vivo. Our results showed that ELS could induce schizophrenia-like behaviors and M1 microglial activation, NAD+ decline, lower expression of SIRT3, and increased acetylated superoxide dismutase 2 expression at the adult stage. NAD+ supplement or HNK administration could block this process and normalize the behavioral alterations of the MS animals. 3-TYP administration in the control group and the NAM-treated MS rats caused M1 microglial activation and cognitive deficits. Our results demonstrated that SIRT3 mediated the stabilizing effect of NAD+ on normalizing M1 microglial activation and behavioral phenotypes in MS rats.

Resveratrol attenuates early life stress induced depression in rats: Behavioural and neurochemical evidence

PurposeMaternal deprivation (MD), a severe naturalistic type of stress in the early postnatal days, is a well-established model of early life stress (ELS) that models juvenile adversity and may result in significant depressive disease in adults. In order to analyze the behavioural, brain monoamine level and HPA axis dysregulations caused by ELS and to determine whether Resveratrol (Res) could counteract these effects, Wistar rat pups were subjected to the MD paradigm, which simulated the consequences of depression. MethodsThe pups on their postnatal day 1–10 were divided in 5 groups (n = 8); nondeprived (ND), maternally deprived (DC), standard fluoxetine (FLX) (5 mg/kg i.p), Res (20, 40 mg/kg i.p). Excluding the ND group, other pups were separated from dam for 3hr/day from day 1 to 10th day. Treatment was initiated from 50th day and was given for 12 days. The behaviour parameters light/dark test, sucrose preference, and resident intruder test were employed. Serum cortisol levels, brain antioxidant activity, monoamine levels and neuronal morphology in the hippocampus were assessed. ResultsThe MD rats showed altered behaviour, including more light–dark transitions, less desire for sucrose, and lower attack latencies. MD influenced the release of serum cortisol and interfered with monoamine, antioxidant levels as well as reduced Nissl bodies in the hippocampus. Treatment with Res led to improved behavioural functions also restored monoamine levels, reduced cortisol release, oxidative stress and prevented histopathological alterations in the rat hippocampus. ConclusionRes showed neuroprotective effects by improving the brain antioxidants and monoamine levels and HPA axis dysregulation and thus improves MD induced depression like behaviour in Wistar rats.

Metabolic cross‐talk between astrocytes and neurons: implications for Alzheimer’s disease

AbstractBackgroundConverging evidence indicates that declining brain energetics contributes to the initiation and progression of neurodegenerative disorders, including Alzheimer’s disease (AD). Upstream events often characterizing AD include hypometabolism of glucose and oxygen in the brain, which may cause mitochondrial dysfunction, energetic failure, and oxidative stress (Moreira et al., 2010). Astrocytes, which have a main role in supporting neuronal function and metabolism, might contribute to the development of neurodegenerative diseases. By modulating the release of gliotransmitters (e.g. glutamate), they provide a fine control of neuronal metabolism and cell‐to‐cell communication, both of them involved in AD (Monterey et al., 2021). Impaired astrocytes metabolism may incite an inflammatory response that, in turn, negatively impacts neuronal viability.MethodPrimary rat cortical astrocytes were isolated from the cortex of Wistar rat pups. A metabolic stress was induced by using glyceraldehyde (GA) (Magi et al., 2021). We evaluated cell viability, the release of tumor necrosis factor alpha (TNFα) and the involvement of the nuclear factor‐kappa B (NF‐kB) pathway. In order to check the glutamate turnover, we analyzed the expression of GLAST and GLT1 and we evaluated the ATP content after glutamate exposure under physiological conditions. We also explored whether damaged astrocytes could elicit neuronal death in a co‐culture setting.ResultExposure to increasing concentrations of GA caused a concentration‐dependent cell injury in rat cortical astrocytes. GA (1mM) started to induce a significant cell damage after 24 h. After 48 h, GA exposure activated the NF‐kB pathway and the release of TNFα. GA treatment significantly increased GLAST and GLT1 levels after 48h. When injured astrocytes were co‐cultured with cortical neurons, neurons viability significantly decreased. Under physiological condition, glutamate significantly improved intracellular ATP levels after 4 and 8 hours of treatment, paving the way to the evaluation of its action in metabolic‐impaired setting, as induced by GA treatment.ConclusionOur results suggest that metabolically impaired astrocytes may damage neighboring neurons, suggesting astrocytes involvement in the pathophysiological development and progression of AD. Therefore, astrocytes can be considered a key therapeutic and neuroprotective target for future studies.

Capsaicin alleviates neuronal apoptosis and schizophrenia-like behavioral abnormalities induced by early life stress

Early life stress (ELS) is associated with the later development of schizophrenia. In the rodent model, the maternal separation (MS) stress may induce neuronal apoptosis and schizophrenia-like behavior. Although the TRPV1 agonist capsaicin (CAP) has been reported to reduce apoptosis in the central nervous system, its effect in MS models is unclear. Twenty-four hours of MS of Wistar rat pups on postnatal day (PND9) was used as an ELS. Male rats in the adult stage were the subjects of the study. CAP (1 mg/kg/day) intraperitoneal injection pretreatment was undertaken before behavioral tests for 1 week and continued during the tests. Behavioral tests included open field, novel object recognition, Barnes maze test, and pre-pulse inhibition (PPI) test. MS rats showed behavioral deficits and cognitive impairments mimicking symptoms of schizophrenia compared with controls. MS decreased the expression of TRPV1 in the frontal association cortex (FrA) and in the hippocampal CA1, CA3, and dentate gyrus (DG) regions compared with the control group resulting in the increase of pro-apoptotic proteins (BAX, Caspase3, Cleaved-Caspase3) and the decrease of anti-apoptotic proteins (Bcl-2). The number of NeuN++TUNEL+ cells increased in the MS group in the FrA, CA1, CA3, and DG compared with the control group. Neuronal and behavioral impairments of MS were reversed by treatment with CAP. Exposure to ELS may lead to increased neuronal apoptosis and impaired cognitive function with decreased TRPV1 expression in the prefrontal cortex and hippocampus in adulthood. Sustained low-dose administration of CAP improved neuronal apoptosis and cognitive function. Our results provide evidence for future clinical trials of chili peppers or CAP as dietary supplements for the reversal treatment of schizophrenia.

Effects of Nicotine Administration in an Enriched Environment on the Behavior of Male MK-801-Exposed Rats.

Smoking is more common in patients with schizophrenia than in healthy populations. Some controversial hypotheses connect the disease with the high prevalence of smoking. Moreover, environmental factors affect the severity of the positive and negative symptoms of schizophrenia. The current study aimed to assess the effect of enriched environment (EE) and nicotine on the MK-801 animal model of schizophrenia. Male Wistar rat pups randomly received saline or MK-801 (dose:1 mg/kg) for five days from the sixth postnatal day (P) until the tenth. The pups were placed in EE or standard cages (SCs) after weaning (P21). Morris water maze (MWM) was used to assess spatial learning and memory. The rats received 0.6 mg/kg nicotine twice for three days at the end of the second month and were examined in an open-field box and three-chamber social interaction test. MK-801 rats' behaviors were the same as those of the saline rats when they were exposed to nicotine. No positive effects of EE were observed when the animals were exposed to nicotine. The results suggested that nicotine decreased schizophrenia-like symptoms and covered the positive effects of EE.

Effect of Blackberry Juice Consumption by Pregnant Rats on Brain Length and Cell Density of Dentate Gyrus in Male Wistar Pups.

The aim of this work was to evaluate the effect of blackberry juice consumption during pregnancy on the length of the brain, as well as on the cell density of the dentate gyrus in Wistar rat pups. Pregnant rats were divided into three groups: control (C), fed with standard diet and water ad libitum; BJ1, which received blackberry juice containing polyphenols (7.8 mg/kg) and anthocyanins (1.9 mg/kg); and BJ2, receiving blackberry juice containing polyphenols (9.3 mg/kg) and anthocyanins (3.54 mg/kg). On postnatal day 0, pups per litter, body weight, and length were measured, and cells in the dentate gyrus of male pups were quantified. Maternal body weight and pups per litter were statistically equal across experimental groups during pregnancy. Pups in BJ1 and BJ2 groups showed an increase in body weight (20%) and length (5%) when comparing to controls. An increase in brain length was observed in BJ2 group (8%) as compared to the control. A significant increase in the number of cells/mm2 was observed in the dentate gyrus of the offspring in BJ1 (21.8%) and BJ2 (23.7%) groups when compared to the control group. Given the above, blackberry juice may be considered a potential functional food during pregnancy, while further research on prenatal and postnatal development must be done.

Impact of Early Arsenic Exposure on the Mineral Content and Oxidative Status of the Liver and Kidney of Pubescent and Adult Rats.

This study evaluated the effect of prepubertal arsenic exposure in the liver and kidney of pubescent rats and their reversibility 30 days after arsenic withdrawal. Male pups of Wistar rats (21 days old) were divided into two groups (n = 20/group): control animals received filtered water, and exposed rats received 10 mg L-1 arsenic from postnatal day (PND) 21 to PND 51. The liver and kidney of 52 days old rats (n = 10/group) were examined to investigate the effects of arsenic on micromineral content, antioxidant enzyme activity, histology, and biochemistry parameters. The other animals were kept alive under free arsenic conditions until 82 days old and further analyzed by the same parameters. Our results revealed that 52-day-old rats increased arsenic content in their liver and arsenic and manganese in their kidney. In those animals, glycogen and zinc content and catalase activity were reduced in the liver, and the selenium content decreased in the kidney. Thirty days later, arsenic reduced the manganese and iron content and SOD and CAT activity in the liver of 82-day-old rats previously exposed to arsenic, while glycogen and selenium content decreased in their kidney. In contrast, PND 82 rats exhibited higher retention of copper in the liver, an increase in iron and copper content, and CAT and GST activity in the kidney. Significant histological alterations of liver and kidney tissues were not observed in rats of both ages. We conclude that arsenic-induced toxicity could alter differently the oxidative status and balance of trace elements in pubertal and adult rats, demonstrating that the metalloid can cause effects in adulthood.

Bumetanide Attenuates Cognitive Deficits and Brain Damage in Rats Subjected to Hypoxia-Ischemia at Two Time Points of the Early Postnatal Period.

Neonatal hypoxia-ischemia (HI) is one of the main causes of tissue damage, cell death, and imbalance between neuronal excitation and inhibition and synaptic loss in newborns. GABA, the major inhibitory neurotransmitter of the central nervous system (CNS) in adults, is excitatory at the onset of neurodevelopment and its action depends on the chloride (Cl-) cotransporters NKCC1 (imports Cl-) and KCC2 (exports Cl-) expression. Under basal conditions, the NKCC1/KCC2 ratio decreases over neurodevelopment. Thus, changes in this ratio caused by HI may be related to neurological disorders. The present study evaluated the effects of bumetanide (NKCC cotransporters inhibitor) on HI impairments in two neurodevelopmental periods. Male Wistar rat pups, 3 (PND3) and 11 (PND11) days old, were submitted to the Rice-Vannucci model. Animals were divided into 3 groups: SHAM, HI-SAL, and HI-BUM, considering each age. Bumetanide was administered intraperitoneally at 1, 24, 48, and 72h after HI. NKCC1, KCC2, PSD-95, and synaptophysin proteins were analyzed after the last injection by western blot. Negative geotaxis, righting reflex, open field, object recognition test, and Morris water maze task were performed to assess neurological reflexes, locomotion, and memory function. Tissue atrophy and cell death were evaluated by histology. Bumetanide prevented neurodevelopmental delay, hyperactivity, and declarative and spatial memory deficits. Furthermore, bumetanide reversed HI-induced brain tissue damage, reduced neuronal death and controlled GABAergic tone, maintained the NKCC1/KCC2 ratio, and synaptogenesis close to normality. Thereby, bumetanide appears to play an important therapeutic role in the CNS, protecting the animals against HI damage and improving functional performance.

Scalp acupuncture improves disordered behavior in the valproic acid-induced rat model of autism: 头针改善丙戊酸钠诱导的孤独症大鼠行为学及其机制研究

ObjectivesTo investigate the underlying mechanisms of scalp acupuncture treatment (SAT) on autism spectrum disorder (ASD). MethodsThirty male Wistar rat pups that had been prenatally exposed to valproic acid sodium (VPA) were randomly divided into the VPA, VPA+acupoint, and VPA+ non-acupoint groups using the random number table method, with 10 rats in each group. Ten pups who had been prenatally exposed to saline were assigned to the control group (CG). There was no intervention in either the control or VPA groups. In the acupoint group, “Shenting (GV24),” bilateral “Benshen (GB13)” were manipulated. In the non-acupoint group, the area below the costal space was stimulated. Acupuncture stimulation lasted for 40 min, with manual twisting of the needles every 10 min, 5 days/week, with 2 days of rest per week, for a total duration of 4 weeks. After the corresponding treatments, behavioral tests (including the open field, social interaction, and Morris water maze tests) were performed to evaluate the therapeutic effects. RT-PCR and Western blotting were performed to detect the expression of neuregulin 1 (NRG1)/ErbB4. ResultsIn the open field test, the activity time spent in the central area in the VPA+acupoint group was significantly longer than that in the VPA group and VPA+ non-acupoint group (both P<0.05). The total length in the VPA+acupoint group was significantly longer than that in the VPA group (P<0.05). The number of bouts in the central area of the VPA+acupoint group was significantly higher than that of the VPA group (P<0.05). In session I of social interaction test, all experimental rats spent more time interacting with stranger 1 (all P<0.05). In session II, the CG and VPA+acupoint groups rats showed more interest in searching for new strangers, but the VPA+non-acupoint group spent more time interacting with stranger 1 than with stranger 2(all P<0.05). In the Morris water maze test, compared with the VPA group, the latency of the VPA+acupoint group was shorter (day 2, 3, 4, 5, P<0.05); compared with VPA+acupoint group, the latency of the VPA+non-acupoint group was longer (day 2, 4, P<0.05). The mean distance in the VPA+acupoint group was shorter than that in the VPA group (day 3, 5, P<0.05). The platform quadrant time of the VPA+non-acupoint group was significantly shorter than that of the VPA+acupoint group (P<0.05) (day 6). The VPA+acupoint group had more platform crossings than the VPA group (P<0.05), and the VPA+ non-acupoint group had fewer platform crossings than the VPA+acupoint group (P<0.05) (day 6). After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly increased than those in the VPA group (both P<0.05) in the medial prefrontal cortex (mPFC). The expression levels of NRG1 and ErbB4 proteins in VPA+non-acupoint group were significantly less than those in the VPA+acupoint group (both P<0.05) in the mPFC. After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly higher than those in the VPA group (both P<0.05) in the hippocampus. The expression levels of NRG1 and ErbB4 proteins in the VPA+non-acupoint group were significantly lower than those in the VPA+acupoint group (both P<0.05) in the hippocampus. After SAT, the expression levels of mRNA levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+acupoint group were significantly higher than those in the VPA group (all P<0.05). The mRNA expression levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+ non-acupoint group were significantly lower than those in the VPA+acupoint group (all P<0.05). ConclusionsOur results suggest that SAT can improve ASD-like behaviors in young rats in a VPA model of ASD. This may be related to its function in upregulating the expression of protein and gene of NRG1 and its receptor ErbB4 in the mPFC.

Creatine supplementation increases postnatal growth and strength and prevents overexpression of pro-inflammatory interleukin 6 in the hippocampus in an experimental model of cerebral palsy

ABSTRACT Objectives: The aim of this study was thus to evaluate the effect of Cr supplementation on morphological changes and expression of pro-inflammatory cytokines in the hippocampus and on developmental parameters. Methods: Male Wistar rat pups were submitted to an experimental model of CP. Cr was administered via gavage from the 21st to the 28th postnatal day, and in water after the 28th, until the end of the experiment. Body weight (BW), food consumption (FC), muscle strength, and locomotion were evaluated. Expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) were assessed in the hippocampus by quantitative real-time polymerase chain reaction. Iba1 immunoreactivity was assessed by immunocytochemistry in the hippocampal hilus. Results: Experimental CP caused increased density and activation of microglial cells, and overexpression of IL-6. The rats with CP also presented abnormal BW development and impairment of strength and locomotion. Cr supplementation was able to reverse the overexpression of IL-6 in the hippocampus and mitigate the impairments observed in BW, strength, and locomotion. Discussion: Future studies should evaluate other neurobiological characteristics, including changes in neural precursor cells and other cytokines, both pro- and anti-inflammatory.