The Effect of Monosodium Glutamate on Neuronal Signaling Molecules in the Hippocampus and the Neuroprotective Effects of Omega-3 Fatty Acids.

Monosodium glutamate (MSG) is a flavoring substance added to many ready-to-eat foods and has known neurotoxic effects. This study was performed in order to examine the potential toxic effect of MSG on neurons in various regions of the hippocampus in prepubertal rats. It also investigated the protective effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on brain-derived neurotropic factor (BDNF), n-methyl-d-aspartate receptor (NMDA-R), and neuropeptide-Y (NPY) expression in the brain, using immunohistochemical and biochemical methods. Six female prepubertal Wistar albino rats were used in each group. Group 1, the control group, received 0.9% saline solution subcutaneously (sc) on days 1, 3, 5, 7, and 9. Group 2 received 4 mg/g MSG sc on days 1, 3, 5, 7, and 9. Group 3 received MSG + EPA (4 mg/g sc on days 1, 3, 5, 7, and 9. Oral 300 mg/kg for 9 d), while Group 4 received MSG + DHA (4 mg/g sc on days 1, 3, 5, 7, and 9 and 300 mg/kg orally for 9 d, respectively). At the end of the ninth day the hippocampal regions of the brain were removed and either fixed for immunohistochemical staining or stored at -80 °C for biochemical parameter investigation. BDNF, NMDA-R, and NPY expression results were evaluated using immunohistochemistry and an enzyme-linked immunosorbent assay. According to our findings, neurons in the control group hippocampal CA1 and DG regions exhibited strong BDNF, NPY, and NMDA-R reactions, while an expression in both regions decreased in the MSG group (p < 0.00). However, in the MSG-EPA and MSG-DHA groups, BDNF, NPY, and NMDA-R immunoreactions in neurons in the same region were similar to those of the control group (p = 0.00). No significant difference was observed in terms of expression in hippocampal neurons between the MSG-EPA and MSG-DHA groups (p > 0.00). In conclusion, since MSG caused a decrease in BDNF, NMDA-R, and NPY neural signaling molecules in the CA1 and DG regions of the hippocampus of prepubertal rats compared to the control group, care is required over the consumption of MSG, since it may affect memory-related neurons in these age groups. In addition, we concluded that the use of omega-3 fatty acids such as EPA and DHA in addition to MSG may protect against the neurotoxic effects of MSG.

Long term neuroprotective effects of acute single dose MK-801treatment against traumatic brain injury in immature rats.

Because brain development continues during adolescence, childhood trauma is a major health problem in pediatric ages. It is known traumatic brain injury (TBI) results in damage in hippocampal and cortical areas of the brain and impairs cognitive functions. The study aims to investigate the long-term effects of MK-801 (dizocilpine), an N-methyl d-aspartate (NMDA) receptor antagonist, on hippocampal damage, locomotor activity, and cognitive functions following TBI in immature rats. MK-801 (1 mg/kg) was injected intraperitoneally immediately after TBI. Thirty-seven litters were randomly allocated into three groups at 7 days (P7) of postnatal age: a control group, a trauma group, and an MK-801 treatment group. The control group received no treatment; the trauma group received saline as vehicle control for the MK-801 group and the MK-801 group received a single dose of 1 mg/kg MK-801 immediately after TBI. Hippocampal damage was examined by Hematoxylin-Eosin staining. Brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), NMDA-R, and glial fibrillar acidic protein (GFAP) immunohistochemistry and, BDNF, NGF, and NMDA-R ELISA protein levels were evaluated 125 days after trauma. Histopathological and immunohistochemical evaluations showed that treatment with MK-801 significantly ameliorated the trauma-induced hippocampal neuron loss and increased BDNF, NGF, NMDA-R, GFAP expressions in CA1, CA3, and DG hippocampal regions. Additionally, treatment with MK-801 decreased anxiety and increased hippocampus-dependent memory of animals subjected to brain injury after TBI. These results show that acute treatment of MK-801 has a neuroprotective role against trauma-induced hippocampal neuron loss and associated cognitive impairment in rats.

Serum Nesfatin-1 Levels in Girls with Idiopathic Central Precocious Puberty.

OBJECTIVE: Nesfatin-1, an anorexigenic neuropeptide, is expressed mainly in the central nervous system and in some peripheral tissues. The role of nesfatin-1 in energy balance has been investigated. Despite the suggestion of a role for nesfatin-1 in reproductive function, data are limited on the role of nesfatin-1 in human puberty. METHODS: The aim of this study was to investigate the following: i) the role of nesfatin-1 in puberty, and ii) relationship between nesfatin-1 and anthropometric measurements and gonadotropin levels in girls with idiopathic central precocious puberty (CPP). Twenty-four girls with CPP (7.68±1.02 years) and 20 female, prepubertal, healthy controls (7.48±0.88 years) were enrolled in the study. All patients with CPP were treated by the intramuscular administration of leuprolide acetate at a daily dose of 3.75 mg for 28 days. Nesfatin-1 was measured before and during treatment. RESULTS: There was no difference in serum nesfatin-1 levels in girls with CPP and healthy controls [5.67 (2.5-20.6) mmol/L and 5.75 (2.51-9.64) mmol/L], respectively. There was a negative correlation between nesfatin-1 levels and body weight and body mass index-standard deviation score (p=0.01, r=-0.83; p=0.025, r=-0.81, respectively). No correlation was found between nesfatin-1 and gonadotropin, estradiol levels, uterine length or endometrial thickness. CONCLUSION: The results of this study suggest that there are no differences between girls with CPP and healthy, prepubertal girls regarding nesfatin-1 levels.

The role of resveratrol on full - Thickness uterine wound healing in rats.

OBJECTIVE: Healing of the uterus after cesarean section and myomectomy operation is clinically important. In this study, we aimed to investigate the effects of resveratrol (3,5,4'-o-trihydroxystilbene) on the wound healing process of the uterus in rats treated with resveratrol following full thickness injury of the uterus. MATERIALS AND METHODS: Twenty-one female wistar albino rats were divided randomly into three groups (1) control group with no intervention (2) injury group with uterine full thickness injury (3) resveratrol group with uterine full thickness injury and treated with resveratrol. Resveratrol was injected by oral gavage at the doses of 0.5 mg/kg/day for 30 days following uterine full thickness injury. Vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) distributions were assessed using the immunohistochemical methods in tissue and ELISA methods in the tissue homogenate. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were evaluated with colorimetric method and malondialdehyde (MDA) levels also were measured using high performance liquid chromatography in the tissue homogenate. The effects of resveratrol on the uterine histology also were evaluated histologically with the light microscopy. RESULTS: Histological evaluation and immunohistochemical evaluations showed that treatment with a resveratrol significantly increased the thickness of the uterine wall and VEGF expression and decreased expression PDGF during wound healing. Biochemically, GPx and SOD activities were increased significantly after treatment with resveratrol. Additionally, resveratrol administration decreased MDA levels. CONCLUSION: These results showed that the antioxidant effects of resveratrol has been shown to have a positive influence on wound healing of the uterus.

Perilipin polymorphisms are risk factors for the development of obesity in adolescents? A case-control study.

BACKGROUND: The variations in perilipin gene (PLIN) were previously associated with obesity. We examined the association of polymorphisms at the PLIN locus in adolescents with obesity and their connection with serum adipokines. METHODS: A total of 308 children (206 obese, 66.8% and 102 healthy control, 33.2%) between the ages of 10-18 years were included into the study. PLIN gene analysis [PLIN 1, PLIN 4, PLIN 6, PLIN 5'UTR-1234 C > G and PLIN 10171 A/T] were studied by Real Time-PCR. Serum leptin, adiponectin, resistin and ghrelin levels were studied by ELISA method in both groups and their link with perilipin polymorphisms were analyzed. RESULTS: Serum leptin level was found significantly high in obese adolescents. Other adipokine levels were similar in both groups. The incidence of PLIN 1, PLIN 4, PLIN 5'UTR-1234 C > G and PLIN 10171 A/T minor and major alleles was similar in both groups. PLIN 6 T/T allele was determined significantly high in obese adolescents compared to that of control group. No correlation was detected between perilipin polymorphism and serum levels of adipokines. CONCLUSION: The PLIN 6 polymorphism of the perilipin gene may influence the risk of the obesity during adolescence. TRIAL REGISTRATION: Retrospectively registered.

Evaluation of the relationship between serum adropin levels and blood pressure in obese children.

BACKGROUND: The prevalence of obesity and related cardiovascular comorbodities is increasing rapidly. Adipokines play a major role in the pathogenesis of obesity-related inflammation and hypertension. AIM: The aim of this study was to evaluate the serum adropin levels in obese children and to determine the relationship between adropin levels and blood pressure (BP) in the pediatric age group. METHODS: Forty obese children (mean age: 12.5 ± 2.5 years; male/female ratio: 18/22) and 15 healthy controls (mean age: 15 ± 3.14 years; male/female ratio: 5/15) were included in the study. Serum adropin levels, and a number of laboratory and clinical variables were compared. Ambulatory blood pressure monitoring was performed on obese subjects. Relationship between adropin levels and BP variables was examined. RESULTS: Serum adropin levels were significantly lower in obese subjects than in healthy controls (193.56 ± 94 vs. 289 ± 187 pg/mL, p = 0.03). Adropin levels were correlated negatively with body mass index z-score (r = -0.56; p = 0.034). There was no correlation between serum adropin levels and laboratory variables in obese subjects. Five of the patients (12.5%) were nondipper, and nine of the patients (22.5%) had hypertension. There was no significant correlation between serum adropin levels and BP variables. CONCLUSION: Serum adropin levels were significantly lower in obese children; however, there was no correlation between serum adropin levels and BP variables. Further studies are needed to determine the role of adipokines on BP.

Neuroprotective effects of MK-801 against traumatic brain injury in immature rats.

Traumatic brain injury (TBI) is a major health problem in pediatric ages and also has major social, economic, and emotional outcomes, with diverse sequelae in many spheres of everyday life. We aimed to investigate the effect of MK-801, a competitive NMDA receptor antagonist, on hippocampal damage and behavioral deficits on 10-day-old rat pups subjected to contusion injury. The aims of the present study were to determine: (i) the short term effects of MK-801 on hippocampal BDNF, NGF and NMDA receptor immunoreactivity and neuron density in hippocampus (ii) long term effects of MK-801 on cognitive dysfunction following TBI in the immature rats. MK-801, was injected intraperitoneally at the doses of 1mg/kg of body weight immediately after induction of traumatic injury. Hippocampal damage was examined by cresyl violet staining, BDNF, NGF and NMDAR receptor immunohistochemistry on P10 day and behavioral alterations were evaluated using elevated plus maze and novel object recognition tests two months after the trauma. Histopathological and immunohistochemical evaluations showed that treatment with a single dose of 1mg/kg MK-801 (i.p.) significantly ameliorated the trauma induced hippocampal neuron loss and decreased BDNF, NGF and NMDAR expressions in CA1, CA3 and DG hippocampal brain regions. Additionally, treatment with MK-801 ameliorated anxiety and hippocampus dependent memory of animals subjected to trauma. These results show that acute treatment of MK-801 has a neuroprotective role against trauma induced hippocampal neuron loss and associated cognitive impairment in immature rats.

Investigation of adropin and leptin levels in pediatric obesity-related nonalcoholic fatty liver disease.

AIM: Nonalcoholic fatty liver disease (NAFLD) is the accumulation of excess fat in the liver in the absence of alcohol consumption, which is commonly associated with obesity and increased risk of atherosclerosis as well as insulin resistance. Adropin is a recently identified protein encoded by the gene related with energy homeostasis, which is expressed in the liver and the brain and has a role in preventing insulin resistance and obesity. The aim of this study was to investigate the serum adropin and leptin levels in obese adolescents and compare the patients with, and without, NAFLD and with healthy controls. METHODS: Sixty-four obese adolescents (30 with NAFLD, 34 without NAFLD) and 36 healthy controls were enrolled in the study. Serum adropin and leptin levels were evaluated by sandwich enzyme-linked immunosorbent assay. RESULTS: Serum adropin levels were significantly lower in obese children than healthy controls (3.2±1.0 and 9.2±1.2 ng/mL, respectively, p=0.001). Serum leptin levels were significantly higher in patients than in controls (12.4±1.1 and 4.1±3.1 pg/mL, respectively; p=0.000). Serum adropin levels of patients with NAFLD were significantly lower than in patients without NAFLD (2.9±0.5 and 3.5±1.2 ng/mL, respectively; p=0.023) and healthy controls (p=0.000). Logistic regression analysis showed that a decrease in adropin levels was the only independent factor for fatty liver disease in obese adolescents (odds ratio: 3.07, 95% confidence interval 1.14-8.2, p=0.026). Leptin, relative weight and HOMA-IR of the patients were not independent risk factors for NAFLD. CONCLUSIONS: In this study, serum adropin levels were significantly lower in obese adolescents with fatty liver disease compared to patients without fatty liver disease and healthy controls. Lower adropin level was an independent risk factor for NAFLD in obese adolescents in logistic regression analysis. Assessment of serum adropin concentrations may provide a reliable indicator of fatty liver disease in obese adolescents.

Transcutaneous electrical nerve stimulation (TENS) accelerates cutaneous wound healing and inhibits pro-inflammatory cytokines.

The purpose of this study was to evaluate transcutaneous electrical nerve stimulation (TENS) and other common treatment methods used in the process of wound healing in terms of the expression levels of pro-inflammatory cytokines. In the study, 24 female and 24 male adult Wistar-Albino rats were divided into five groups: (1) the non-wounded group having no incision wounds, (2) the control group having incision wounds, (3) the TENS (2 Hz, 15 min) group, (4) the physiological saline (PS) group and (5) the povidone iodine (PI) group. In the skin sections, interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were assessed with enzyme-linked immunosorbent assay and immunohistochemical methods. In the non-wounded group, the expression of IL-1ß, IL-6, and TNF-α signaling molecules was weaker in the whole tissue; however, in the control group, significant inflammatory response occurred, and strong cytokine expression was observed in the dermis, granulation tissue, hair follicles, and sebaceous glands (P < 0.05). In the TENS group, the decrease in TNF-α, IL-1ß, and IL-6 immunoreaction in the skin was significant compared to the other forms of treatment (P < 0.05). Distinctive decreases of pro-inflammatory cytokines observed in the dermis in the TENS group suggest that TENS shortened the healing process by inhibating the inflammation phase.

A Comparison Study of Growth Factor Expression following Treatment with Transcutaneous Electrical Nerve Stimulation, Saline Solution, Povidone-Iodine, and Lavender Oil in Wounds Healing.

This study compared the effects of transcutaneous electrical nerve stimulation (TENS), saline solution (SS), povidone-iodine (PI), and lavender oil (Lavandula angustifolia) through expression of growth factors in a rat model of wound healing. Six experimental groups were established, each containing 8 rats: a healthy group with no incision wounds, an incision-control group, an incision and TENS group, an incision and SS group, an incision and PI group, and an incision and lavender oil group. Experiments continued for 5 days, after which the skin in the excision area was removed. Tissue concentrations of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF)-A were measured using enzyme-linked immunosorbent assay (ELISA). Tissue expressions of EGF, PDGF-A, and fibroblast growth factor (FGF)-2 were determined using immunohistochemistry. Wound closure progressed more rapidly in the TENS and lavender oil groups than in the control and other study groups. In particular, PDGF-A expressions in the dermis and EGF expression in the epidermis were significantly intense in the TENS group (P < 0.05). In addition, ELISA levels of growth factors such as PDGF-A and EGF were significantly higher in TENS group compared to the control group (P < 0.05). These immunohistochemical and ELISA results suggest that TENS may improve wound healing through increasing growth factors in the dermis and epidermis more than other topical applications.

Evaluation of serum neopterin levels and its relationship with adipokines in pediatric obesity-related nonalcoholic fatty liver disease and healthy adolescents.

AIM: Nonalcoholic fatty liver disease (NAFLD) is associated with inflammation and increased risk of atherosclerosis. Neopterin is regarded as a biochemical marker of cell-mediated immunity, which is secreted by monocytes and macrophages, mainly in response to interferon-gamma. The aim of the present study was to investigate the serum neopterin levels in obese adolescents and compare the neopterin levels in patients with and without NAFLD and also with healthy controls. The second aim of the study was to research the possible relationship between neopterin levels and adipokines (leptin, adiponectin, resistin, and ghrelin). METHODS: Ninety-three obese adolescents (39 with NAFLD, 54 without NAFLD) and 55 healthy controls were enrolled in the study. Serum levels of neopterin and adipokines were measured with high-performance liquid chromatography and sandwich enzyme-linked immunosorbent assay, respectively. RESULTS: Serum neopterin levels were significantly higher in patients with NAFLD (3.20 ± 0.09 nmol/L) than in their healthy peers (2.91 ± 0.08 nmol/L) (p=0.020). Neopterin levels were positively correlated with leptin levels in obese patients (r=0.380, p<0.001) and in the group comprising all individuals (r=0.206, p<0.05). There was no correlation between neopterin concentrations and relative weight, alanin aminotransferase, adiponectin, resistin, and ghrelin levels. CONCLUSION: The serum neopterin levels were significantly higher in obese adolescents with fatty liver disease compared to controls, and this may be related to increased cell-mediated immunity in fatty liver disease.

The effect of lipoic acid on wound healing in a full thickness uterine injury model in rats.

Aim of this study was to investigate the effects of lipoic acid on uterine wound healing by immunohistochemical and biochemical assay in a rat uterine horn model with full thickness injury. Thirty-two female Wistar albino rats were randomised into five groups: Control group, with no intervention; uterine scar group 15days (US15d), uterine scar group 15 days + alpha lipoic acid (ALA) (US15d + ALA), uterine scar group 30 days (US30d) and uterine scar group 30 days + ALA (US30 days + ALA). After uterine incision 100 mg/kg of ALA was administered by oral gavage for either 15 or 30 days. Vascular endothelial growth factor (VEGF) and alpha smooth muscle actin (α-SMA) distribution were evaluated by immunohistochemical methods in tissue and ELISA methods in tissue homogenate. The percentage of α-SMA positive area in US15d + ALA and US30d + ALA groups was significantly higher than US15 and US30d groups. The percentage of VEGF positive area in US15d + ALA group was significantly higher than US15d group and US30d + ALA group was significantly higher than US30d group. Biochemically, α-SMA was significantly higher in the US15d + ALA group when compared to US15d group and higher in US30d + ALA group when compared to US30d group. VEGF was significantly higher in US15d + ALA and US30d + ALA groups when compared to US15 and US30d groups. In conclusion, ALA was found to be effective in enhancing wound healing in uterine full thickness injury.

Effects of resveratrol on incisional wound healing in rats.

PURPOSE: The objective of this study was to investigate the effect of resveratrol on the healing process after midline laparotomy in rats. METHODS: The study was performed on adult female Wistar-Albino rats. The study group was orally administered 0.5 mg/kg resveratrol once a day for 7 days before the operation until 12 h before surgery and then the treatment was maintained throughout the study. Each rat was anesthetized, and a 4-cm midline laparotomy was performed. Ten animals in each group were sacrificed on postoperative days 7, and 14. A tensile strength analysis was performed, hydroxyproline levels were measured, and the abdominal incision wounds were examined histologically. RESULTS: Resveratrol administration significantly increased the tensile strength of the abdominal fascia, and increased the hydroxyproline levels on postoperative day 14. The acute inflammation scores, collagen deposition scores and the neovascularization scores on postoperative days 7 and 14 were found to be significantly higher in the resveratrol treatment group compared to the control group. The amount of granulation tissue and the fibroblast maturation scores were found to be significantly higher only on postoperative day 14 in the treatment group compared to the control group. CONCLUSION: Our findings show that resveratrol may have a beneficial effect on incisional wound healing.

Lipoic acid decreases peritoneal adhesion formation in a rat uterine scar model.

OBJECTIVE: To investigate the effects of lipoic acid in the prevention of postoperative pelvic adhesions by a visual scoring system and immunohistochemistry in a rat uterine horn model with full thickness injury. MATERIAL AND METHODS: Twenty-eight female Wistar albino rats were randomised into four groups: uterine trauma control, 15 days and 30 days, and uterine trauma + lipoic acid, 15 days and 30 days. A full thickness defect was established by incising a segment of approximately 1.0 cm in length from each uterine horn, leaving the mesometrium intact. Extension and severity of the adhesions in each group were scored by a visual scoring system and evaluated immunohistochemically. RESULTS: Adhesion scores were 2.00±0.81, 2.14±0.69 0.71±0.75, and 0.85±0.69 for extent and 2.28±0.48, 2.14±0.69, 0.85±0.69, and 1.14±0.69 for severity in Groups 1, 2, 3 and 4, respectively. Adhesion extent and severity were significantly less for groups treated by lipoic acid but no difference was observed between long and short administration. Both Vitronectin and u-PAR staining were significantly increased in treatment groups when compared to the control group. CONCLUSION: Lipoic acid was found to be effective in reducing postoperative adhesion formation in a rat model.

The protective effects of resveratrol on human coronary artery endothelial cell damage induced by hydrogen peroxide in vitro.

Oxidative stress is defined as imbalance between the production and destruction of reactive oxygen species. The aim of this study was to investigate whether resveratrol could protect human endothelial cells against hydrogen peroxide damage in vitro. In this in vitro study on human coronary endothelial cells, the effects of resveratrol on the glutathione content in human coronary endothelial cells in vitro were evaluated with high performance liquid chromatography. The effects of resveratrol on protein expression of the glutamate cysteine ligase, glutathione peroxidase and glutathione reductase enzymes were evaluated with the Western blot method. Resveratrol increased the reduced glutathione contents significantly (p < 0.05). Resveratrol increased protein expression of the glutamate cysteine ligase, glutathione peroxidase-1 and glutathione reductase enzymes (p < 0.05). All data supported each other and suggested that resveratrol had a protective effect against human coronary artery endothelial cell damage. It is thought that these results could pave the way to the new therapeutic approaches to protect against oxidative stress that develops in cardiovascular diseases.

Resveratrol reduces matrix metalloproteinase-2 activity induced by oxygen-glucose deprivation and reoxygenation in human cerebral microvascular endothelial cells.

The main pathophysiology in cerebral ischemia is the structural alteration in the neurovascular unit, coinciding with neurovascular matrix degradation. Among the human matrix metalloproteinases (MMPs), MMP-2 and -9, known as gelatinases, are the key enzymes for degrading type IV collagen, which is the major component of the basal membrane that surrounds the cerebral blood vessel. In the present study, we investigated the effects of resveratrol on cytotoxicity, reactive oxygen species (ROS), and gelatinases (MMP-2 and -9) in human cerebral microvascular endothelial cells exposed to 6 hours of oxygen-glucose deprivation and a subsequent 24 hours of reoxygenation with glucose (OGD/R), to mimic ischemia/reperfusion in vivo. Lactate dehydrogenase increased significantly, in comparison to that in the normoxia group. ROS was markedly increased in the OGD/R group, compared to normoxia. Correspondingly, ROS was significantly reduced with 50 µM of resveratrol. The proMMP-2 activity in the OGD/R group showed a statistically significant increase from the control cells. Resveratrol preconditioning decreased significantly the proMMP-2 in the cells exposed to OGD/R in comparison to that in the OGD/R group. Our results indicate that resveratrol regulates MMP-2 activity induced by OGD/R via its antioxidant effect, implying a possible mechanism related to the neuroprotective effect of resveratrol.