Autistic-like behaviors are attenuated by agmatine consumption during pregnancy: Assessment of oxidative stress profile and histopathological changes in the prefrontal cortex and CA1 region of the hippocampus.

Objectives: Due to the crucial role of polyamines during fetal growth and development, we aimed to determine the effect of prenatal administration of agmatine, an endogenous active metabolite of arginine, and a nutritional supplement, on autistic-like behaviors, oxidative-anti-oxidative profile, and histopathological changes of the prefrontal cortex (PFC) and CA1 area of the hippocampus in valproic acid (VPA) model of autism in male rats. Materials and Methods: VPA was injected intraperitoneally on embryonic days (ED) 12.5, and the pregnant rats were gavaged with agmatine between E6.5 to E18.5 (13 days), at doses of 0.001, 0.01, and 0.1 mg/kg. The autism-like behaviors and memory of male pups were analyzed via open-field, three-chamber, and novel object recognition tests. Serum oxidative stress and the histological changes in the PFC and CA1 were assessed at the end of the study. Results: The results suggest that prenatal agmatine reduced autistic-like behaviors by decreasing cell loss in CA1 and PFC. We observed no alterations in superoxide dismutase (SOD) level and total anti-oxidant capacity (TAC) between groups. VPA decreased catalase (CAT) activities, while agmatine decreased malondialdehyde (MDA) activity. Conclusion: Overall, this investigation suggests that agmatine may be a potential candidate for the early treatment and even prevention of appearance of autism symptoms.

Astrocyte response to melatonin treatment in rats under high-carbohydrate high-fat diet.

The involvement of consumption of high-carbohydrate high-fat (HCHF) diet in cognitive impairment is attributed, at least in part, to the activation of astrocytes, which contributes to the development of neuroinflammation, oxidative stress, and subsequent cognitive deficits. This study aimed to assess the influence of melatonin on cognitive impairment and astrogliosis induced by the HCHF diet in rats. Male Wistar rats were fed an HCHF diet for eight weeks to induce obesity and metabolic syndrome. Subsequently, they received oral melatonin treatment for four weeks at doses of 5 mg/kg, 10 mg/kg, and 30 mg/kg, alongside the HCHF diet. Cognitive function was evaluated using the Y-maze test, while the levels of proinflammatory cytokines, oxidative stress, and the number glial fibrillary acidic protein (GFAP) positive cells were assessed in the hippocampi and hypothalamus. The consumption of the HCHF diet resulted in weight gain, hyperlipidemia, impaired glucose tolerance, cognitive decline, neuroinflammation, oxidative stress damage, and astrogliosis in rats. Although melatonin treatment did not demonstrate beneficial effects on blood glucose and lipid metabolism, it improved the impaired working memory caused by the HCHF diet. Melatonin exhibited a dose-dependent reduction of astrogliosis, neuroinflammation, and lipid peroxidation while restored superoxide dismutase in the hippocampus and hypothalamus of HCHF diet-treated rats. These findings provide evidence that melatonin inhibits astrocyte activation, thereby attenuating inflammation and minimizing oxidative stress damage induced by the HCHF diet.

Assessment of melatonin's therapeutic effectiveness against hepatic steatosis induced by a high-carbohydrate high-fat diet in rats.

Several studies have demonstrated the protective effects of melatonin against metabolic diseases, such as liver steatosis. However, its therapeutic effects have received less scrutiny. The present study aimed to explore melatonin's therapeutic effectiveness in treating non-alcoholic fatty liver disease (NAFLD) induced by a high-carbohydrate high-fat (HCHF) diet in rats. The NAFLD was developed in male Wistar rats using an HCHF diet for 8 weeks. Afterward, they were given melatonin orally for four weeks at doses of 5 mg/kg, 10 mg/kg, and 30 mg/kg, along with the HCHF diet. In addition, six age-matched healthy rats received the highest dose of melatonin (30 mg/kg) for the same duration. Rats on the HCHF diet exhibited obesity, dyslipidemia, hyperglycemia, glucose intolerance, insulin resistance, inflammation, oxidative stress, and liver injury (steatosis). Melatonin treatment at 10 mg/kg and 30 mg/kg reduced body weight, adiposity index, oxidative damage, and inflammation but did not affect impaired glucose metabolism induced by the HCHF diet. Meanwhile, the highest dose of melatonin (30 mg/kg) reduced the liver steatosis index in HCHF rats but caused mild liver damage in healthy rats. In conclusion, using melatonin demonstrated positive outcomes in treating NAFLD induced by the HCHF diet in rats, with no noteworthy effects observed in healthy rats. A moderate dosage of 10 mg/kg of melatonin proved to be a safer and more efficient method for reducing HCHF diet-induced NAFLD in rats. Higher melatonin doses should be cautiously administered due to potential disruptions in lipid metabolism and the risk of liver complications.

Agmatine improves liver function, balance performance, and neuronal damage in a hepatic encephalopathy induced by bile duct ligation.

INTRODUCTION: In the current study, we investigate whether oral administration of agmatine (AGM) could effectively reduce motor and cognitive deficits induced by bile duct ligation (BDL) in an animal model of hepatic encephalopathy (HE) through neuroprotective mechanisms. METHODS: The Wistar rats were divided into four groups: sham, BDL, BDL+ 40 mg/kg AGM, and BDL+ 80 mg/kg AGM. The BDL rats were treated with AGM from 2 weeks after the surgery for 4 consecutive weeks. The open field, rotarod, and wire grip tests were used to assess motor function and muscle strength. The novel object recognition test (NOR) was performed to evaluate learning and memory. Finally, blood samples were collected for the analysis of the liver markers, the animals were sacrificed, and brain tissues were removed; the CA1 regions of the hippocampus and cerebellum were processed to identify apoptosis and neuronal damage rate using caspase-3 immunocytochemistry and Nissl staining. RESULTS: The serological assay results showed that BDL severely impaired the function of the liver. Based on histochemical findings, BDL increased the neuronal damage in CA1 and Purkinje cells, whereas apoptosis was significantly observed only in the cerebellum. AGM treatment prevented the increase of serum liver enzymes, balance deficits, and neuronal damage in the brain areas. Apoptosis partially decreased by AGM, and there were no differences in the performance of animals in different groups in the NOR. CONCLUSIONS: The study suggests AGM as a potential treatment candidate for HE because of its neuroprotective properties and/or its direct effects on liver function.

In utero and postnatal exposure to Foeniculum vulgare and Linum usitatissimum seed extracts: modifications of key enzymes involved in epigenetic regulation and estrogen receptors expression in the offspring's ovaries of NMRI mice.

BACKGROUND: Early-life exposure to exogenous estrogens such as phytoestrogens (plant-derived estrogens) could affect later health through epigenetic modifications. Foeniculum vulgare (fennel) and Linum usitatissimum (flax) are two common medicinal plants with high phytoestrogen content. Considering the developmental epigenetic programming effect of phytoestrogens, the main goal of the present study was to evaluate the perinatal exposure with life-long exposure to hydroalcoholic extracts of both plants on offspring's ovarian epigenetic changes and estrogen receptors (ESRs) expression level as signaling cascades triggers of phytoestrogens. METHODS: Pregnant mice were randomly divided into control (CTL) that received no treatment and extract-treated groups that received 500 mg/kg/day of fennel (FV) and flaxseed (FX) alone or in combination (FV + FX) during gestation and lactation. At weaning, female offspring exposed to extracts prenatally remained on the maternal-doses diets until puberty. Then, the ovaries were collected for morphometric studies and quantitative real-time PCR analysis. RESULTS: A reduction in mRNA transcripts of the epigenetic modifying enzymes DNMTs and HDACs as well as estrogen receptors was observed in the FV and FX groups compared to the CTL group. Interestingly, an increase in ESRα/ESRß ratio along with HDAC2 overexpression was observed in the FV + FX group. CONCLUSION: Our findings clearly show a positive relationship between pre and postnatal exposure to fennel and flaxseed extracts, ovarian epigenetic changes, and estrogen receptors expression, which may affect the estrogen signaling pathway. However, due to the high phytoestrogen contents of these extracts, the use of these plants in humans requires more detailed investigations.

L-carnitine attenuates acoustic startle reflex dysfunction in adult male rats exposed to mancozeb.

The fungicide mancozeb increases oxygen-free radicals in the central nervous system. As an antioxidant, L-carnitine protects DNA and cell membranes from damage caused by oxygen-free radicals. The present study investigated how L-carnitine affected the acoustic startle response (ASR) in rats exposed to mancozeb. In this experimental study, male Wistar rats were gavaged orally with mancozeb (500, 1000, and 2000 mg/kg), L-carnitine (100, 200, and 400 mg/kg), or L-carnitine (200 mg/kg) + mancozeb (500 mg/kg) three times in 1 week. In the sham group, saline (0.9%, 10 mL/kg) was gavaged at a volume equivalent to that of the drugs. The control group did not receive any treatment. The results showed that locomotor activity and the percentage of prepulse inhibition in the mancozeb groups decreased compared to the sham group while these parameters increased in the L-carnitine group (200 mg/kg) compared to sham rats. In conclusion, mancozeb may increase the risk factor for cognitive diseases such as schizophrenia in people exposed to it while pretreatment with L-carnitine can attenuate the toxic effect.

Serum Scavenging Capacity and Folliculogenesis Impact following Flaxseed Consumption in the First-Generation Mice Pups.

Flaxseed is a source of antioxidants utilized for female infertility treatment in traditional medicine. This study investigated the effects of flax hydroalcoholic extract and flaxseeds during prenatal and postnatal (PND) periods on folliculogenesis and serum total antioxidant capacity (TAC). Pregnant NMRI mice received 500 and 1000 mg/kg of flax extract (LE) and the same doses of flaxseed (LS). Female pups received the same regimen for 56 days. The body, ovarian morphometry, follicle development, and TAC levels were evaluated. The ovarian weight significantly increased in the LE1000 group compared to the LS500 group. The LE500 group had a considerably lower number of primary and antral follicles compared to the CTL and LS1000 groups. The number of antral follicles significantly increased in the LE1000 group compared to the LS500 and LE500 groups. The number of preovulatory follicles was higher in the LE1000 group. A significant increase in the TAC levels was detected in the LS500, LS1000, and LE1000 groups. LE showed a dose-dependent protective effect on the folliculogenesis in F1, which is more evident with the dosage of 1000 mg/kg. This could be related to the strongest antioxidant property of LE1000, as shown by the highest levels of TAC.

Evaluation of expression and serum concentration of anti-Mullerian hormone as a follicle growth marker following consumption of fennel and flaxseed extract in first-generation mice pups.

BACKGROUND: The aim of the present study was to assess the expression and serum level of AMH in first-generation female mice pups following fennel and flaxseed consumption. METHODS: Twenty pregnant NMRI mice were allocated into four groups including control (CTL), fennel (FV), flaxseed (LU) and FV+ LU. Sixty-four female offsprings after lactation period, received the same regimen as their mothers for 56 and 240 days. The ovarian follicles development, serum concentration of AMH, as well as gene and protein expression of AMH were evaluated in the female offsprings at post-natal day 56 (PND56) and 240 (PND240). RESULTS: The number of total growing follicles were raised in the FV group in compression to the all experimental groups. In contrast, LU group showed a marked decrease in their numbers. The highest level of serum AMH was seen in the FV-diet mice, whereas LU negatively affected it. The expression level of AMH also increased in the FV and FV + LU groups, while a reduction was observed in the LU group. As well, IHC data showed that the number of AMH-positive cells in almost ovarian follicles of FV and FV + LU-treated mice was in compared to those of the LU group. CONCLUSIONS: The overall effect of fennel treatment (alone and in combination with flaxseed) on ovary might be maintain primordial follicle storage through increased expression and serum level of AMH.

Hydroalcoholic extract and seed of Foeniculum vulgare improve folliculogenesis and total antioxidant capacity level in F1 female mice offspring.

BACKGROUND: Foeniculum vulgare (fennel) is traditionally suggested for the fertility improvement in Iranian lore due to its antioxidant and phytoestrogen compounds. The present study aimed to investigate the effects of fennel seed and its hydroalcoholic extract on the serum total antioxidant capacity (TAC) and folliculogenesis in offspring exposed to either of the treatments in utero and 56 days after birth (PND 56). METHODS: Pregnant NMRI mice were randomly divided into 5 groups of 7: extract-treated groups received 500 and 1000 mg/kg/day fennel extract (FE), seed-treated groups received 500 and 1000 mg/kg/day fennel seed (FS), and the control group (CTL) received no treatment. The treatments started from pregnancy day 1 and continued until PND 56. Body and right ovary weights and ovary dimensions were recorded. Hematoxylin and eosin stained ovary sections were prepared to calculate the proportion of different follicles. The level of TAC in the serums was also measured by fluorescence recovery after photo bleaching. RESULTS: A marked rise in the body and ovary weights of treated mice was observed compared to the CTL group. The mean number of primordial, primary, pre-antral, and pre-ovulatory follicles as well as corpus luteum size in the treated offspring was significantly higher compared to those of CTL offspring. The atretic follicle number was nonsignificantly lower in either of the treatment groups compared with that in the CTL. However, treatment of animals with 500 mg/kg FE showed a more pronounced effect. Animals in FS500, FE500 and FE1000 groups had a significantly higher level of serum TAC compared to the CTL group. CONCLUSIONS: Fennel extract and seed administration in pregnancy and lactation period improve offspring's folliculogenesis. Higher level of TAC in the serum of offspring might have positively altered the folliculogenesis milieu.

Protective effects of hydro-alcoholic extract of foeniculum vulgare and linum usitatissimum on ovarian follicle reserve in the first-generation mouse pups.

OBJECTIVE: The present study was conducted to investigate the protective effects of fennel and flaxseed during pre- and post-natal period until puberty and menopause on ovarian follicular reserve (OFR). METHODS: Pregnant NMRI mice received fennel (FV, 500 mg/kg/day), flaxseed (LU, 500 mg/kg/day), LU + FV (500 mg/kg/day) and no treatment was given to the controls. Female pups were studied on post-natal-days 1, 56 and 240 (PND1, 56, 240). Ovary weight and diameters, the number of primordial (PF), atretic (AF) and apoptotic (APF) follicles were determined. The expression of Bcl2 and STAT3 (apoptosis-related-genes), micoRNA-125a-5p, and also serum levels of sex hormones were measured. Data were analyzed using one-way ANOVA test. RESULTS: FV and FV + LU groups showed a marked rise in body and ovary weights and diameters as compared to the control group. The number of PF at PND1, PND56, and PND240 increased significantly in the FV and FV + LU groups but decreased in the LU group compared to the control mice. There was a significant reduction in the mean of AF in the FV and FV + LU group and a marked increase in the LU group compared to the controls. Also, more APF were observed in the LU group, whereas less apoptotic follicles were present in the FV group. FSH and estradiol serum levels increased significantly while LH decreased in the FV group. The anti-apoptotic-genes expression and pro-apoptotic microRNA, respectively, increased and decreased in the FV group versus control group. CONCLUSIONS: It can be concluded that fennel alone and in combination with flaxseed could improve OFR during pregnancy, lactation, and afterwards until puberty and menopause.