Bcl-2 In Brain Tissue Research Articles

Insights overview on the possible protective effect of chitosan nanoparticles encapsulation against neurotoxicity induced by carbendazim in rats

Carbendazim (CBZ) contamination of food and water is a principal factor in many negative impacts on public health. Nanoencapsulation of agrochemicals by nontoxic polymers as chitosan nanoparticles (CS-NPs) is one of the most applications of nanotechnology in agriculture. Despite its many advantages, such as it provides controlled release property, more stability and solubility of the active ingredient, it is not authorized to be used in the market because there are no adequate studies on the nano pesticides induced toxicity on experimental animals. So, we aim to study the possible impacts of CBZ-loading CS-NPs on the whole brain of rats and to explain its mechanism of action. 20 male Wistar rats were partitioned into 4 groups as follows: Group (1), normal saline; group (2), 5 mg/kg CS-NPs; group (3), 300 mg/kg CBZ; group (4) 300 mg/kg CS/CBZ-NCs. After 28 days, some neurobehavioral parameters were assessed to all rats then euthanization was done to collect the brain. Our results revealed that CBZ prompted neurotoxicity manifested by severe neurobehavioral changes and a significant increase of MDA with a decrease of GSH and CAT in brain tissue. In addition, there were severe neuropathological alterations confirmed by immunohistochemistry which showed strong bax, GFAP, and TNF-ὰ protein expression in some brain areas. CBZ also induced apoptosis manifested by up-regulation of JNK and P53 with down-regulation of Bcl-2 in brain tissue. Otherwise, encapsulation of CBZ with CS-NPs could reduce CBZ-induced neurotoxicity and improve all studied toxicological parameters. We recommend using CBZ-loading CS-NPs as an alternative approach for fungicide application in agricultural and veterinary practices but further studies are needed to ensure its safety on other organs.

Bone marrow mesenchymal stem cells overexpressing hepatocyte growth factor ameliorate hypoxic-ischemic brain damage in neonatal rats.

ObjectivesHypoxic–ischemic brain damage (HIBD) is a major cause of brain injury in neonates. Bone marrow mesenchymal stem cells (BMSCs) show therapeutic potential for HIBD, and genetic modification may enhance their neuroprotective effects. The goal of this study was to investigate the neuroprotective effects of hepatocyte growth factor (HGF)-overexpressing BMSCs (BMSCs-HGF) against HIBD and their underlying mechanisms.Methods:BMSCs were transfected with HGF using adenoviral vectors. HIBD models were established and then BMSCs were transplanted into the brains of HIBD rats via intraventricular injection. 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure cerebral infarction volumes. In vitro, primary cultured cortical neurons were co-cultured with BMSCs in a Transwell plate system. Oxygen–glucose deprivation (OGD) was applied to imitate hypoxic–ischemic insult, and PD98059 was added to the culture medium to block the phosphorylation of extracellular signal-regulated kinase (ERK). Cell apoptosis was determined using TUNEL staining. The expression of HGF was measured by immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blots. The expression of phosphorylated ERK (p-ERK) and B-cell lymphoma-2 (Bcl-2) was measured by western blots.ResultsHGF-gene transfection promoted BMSC proliferation. Moreover, BMSCs-HGF decreased HIBD-induced cerebral infarction volumes and enhanced the protective effects of the BMSCs against HIBD. BMSCs-HGF also increased expression of HGF, p-ERK, and Bcl-2 in brain tissues. In vitro, BMSC-HGF protected neurons against OGD-induced apoptosis. Inhibition of ERK phosphorylation abolished the neuroprotective effect of BMSCs-HGF against OGD.ConclusionsBMSCs-HGF is a potential treatment for HIBD and that the ERK/Bcl-2 pathway is involved in the underlying neuroprotective mechanism.

Omega-3 Rich Oils Attenuate ADHD-Like Behaviour Induced by Dietary Monosodium Glutamate in Rats.

<b>Background and Objective:</b> Attention Deficit Hyperactivity Disorder (ADHD) is a neurodevelopmental disorder characterized by inattention, hyperactivity and cognitive dysfunction. The present study was designed to examine the possible modulatory effect of Fish, Walnuts or Fenugreek Oils against Attention Deficit Hyperactivity Disorder (ADHD)-like Behavior induced by Monosodium Glutamate (MSG) in Rats. <b>Materials and Methods:</b> Fifty weaning rats were divided into five groups, (each group contain 10 rats) as follows: Group 1: Normal control rats were fed on a balanced diet. Groups from 2-5 rats were fed on a balanced diet+MSG (0.4 g kg<sup></sup><sup>1</sup> diet), Group 2 served as a positive control group whereas group 3, 4 and 5 treated with Fish, Walnuts and Fenugreek oil, respectively, (200 mg kg<sup></sup><sup>1</sup> b.wt.) by intra-gastric tube. Biochemical and behavioural parameters were tested as well as microscopic examination of brain tissue was done. <b>Results:</b> MSG ingestion caused marked disruption in locomotors activity, memory function and brain tissue structure along with significant abnormalities in some bio-markers and reduction in the gene expression level of Bcl-2 in brain tissue. However, treatment with the tested oils showed remarkable effect by reversing the condition. <b>Conclusion:</b> Dietary supplementation with walnut; fenugreek or fish oils at the tested dose could modulate the condition of ADHD in rats.

Neuroserpin extends the time window of tPA thrombolysis in a rat model of middle cerebral artery occlusion.

Tissue-type plasminogen activator (tPA) is characterized as an effective drug for early thrombolytic therapy in acute cerebral infarction (ACI). However, tPA will increase the risk of hemorrhage if it is used beyond the treatment time window. The study aims to explore the effects of neuroserpin (NSP) on the time window of tPA thrombolysis in ACI and the underlying mechanism. The middle cerebral artery occlusion (MCAO) model was constructed in rats, which were randomly divided into six groups: sham operation group, infarction group, 1-hour thrombolysis group, 1-hour thrombolytic + NSP intervention group, 4-hour thrombolytic group, and 4-hour thrombolysis + NSP intervention group. The neurological changes in rats were evaluated by modified neurological severity scores and rota-rod test. The brain edema and cerebral infarction area were evaluated by dry-wet method and triphenyl tetrazolium chloride staining. The blood-brain barrier (BBB) integrity was examined by Evans blue method. The expressions of malondialdehyde, superoxide dismutase, and glutathione peroxidase in brain were also investigated. The expression of caspase-3 and Bcl-2 in brain tissue and apoptosis of neurons were examined by Western blot analysis and toluidine blue staining. tPA thrombolysis significantly attenuated the neurological impairment in rats with MCAO at 1 hour. Conversely, the effect of tPA thrombolysis at 4 hours after MCAO did not significantly help the recovery of neurological function. However, a combination of tPA treatment and NSP treatment at 4 hours after MCAO markedly ameliorated the neurological impairment, cerebral edema, cerebral infarction volume, BBB injury, oxidative stress products, and neuron apoptosis. NSP can probably expand the time window for tPA treatment to reduce neurological impairment in ACI.

Research on mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury in rats through JNK signaling pathway.

To explore the specific mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury (CIRI) in rats through the c-Jun N-terminal kinase (JNK) signaling pathway. A total of 60 male specific pathogen-free Sprague-Dawley rats were randomly divided into sham group (n=20), model group (n=20), and sevoflurane group (n=20). In the sevoflurane group, sevoflurane (2.5%) was inhaled for 60 min at 24 h before the blockage of cerebral blood supply. The CIRI model was established using the suture method in the model group and sevoflurane group, while the right common carotid artery and external carotid artery were separated and ligated only, without suture placement, in the sham group. At 24 h after reperfusion, the neurological deficit score in each group was calculated, the water content in brain tissues in each group was detected based on dry-wet weight ratio, the infarction volume of brain tissues in each group was detected via 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the apoptosis rate of brain cells in each group was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the protein levels of JNK, p-JNK, B-cell lymphoma-2 (Bcl-2), and the Bcl-2 associated X protein (Bax) in brain tissues were determined using Western blotting, and the gene expressions of Bax and Bcl-2 in brain tissues were determined through fluorescence quantitative Polymerase Chain Reaction (qPCR). It was found that the water content in brain tissues and the cerebral infarction volume were significantly increased in the model group compared with those in the sham group (p<0.01, p<0.01), while they were notably decreased in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). The neurological deficit score was significantly higher in the model group than that in the sham group (p<0.01), while it was remarkably lower in the sevoflurane group than that in the model group (p<0.01). According to the results of the TUNEL assay, the model group had an evidently higher apoptosis rate of brain cells than the sham group (p<0.01), while the sevoflurane group had a lower apoptosis rate of brain cells than the model group (p<0.05). Besides, the results of Western blotting revealed that the model group exhibited remarkably increased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and a remarkably decreased protein level of Bcl-2 (p<0.01) compared with the sham group. Sevoflurane group had decreased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and an increased protein level of Bcl-2 (p<0.05) in comparison with the model group. In addition, the gene expression of Bcl-2 significantly declined (p<0.01), and that of Bax remarkably rose (p<0.01) in the model group compared with those in the sham group, while the contrary is the case in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). Sevoflurane can regulate the protein and gene expressions of Bax and Bcl-2 and reduce apoptosis in CIRI by regulating the JNK signaling pathway, thereby exerting a protective effect on brain tissues and improving the symptoms of neurological deficit.

Protective effect of magnesium sulfate on cranial nerves in preeclampsia rats through NF-κB/ICAM-1 pathway.

The aim of this study was to investigate the protective effect of magnesium sulfate (MgSO4) on the cranial nerves of preeclampsia (PE) rats through the nuclear factor-κB (NF-κB)/intercellular adhesion molecule-1 (ICAM-1) pathway. A total of 30 pregnant rats were randomly divided into three groups, including control group, model group, and treatment group, with 10 rats in each group. Systolic blood pressure was measured at 13 d, 15 d, and 19 d. The apoptosis level in brain tissues was detected via Western blotting and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Protein expression of genes was detected using immunohistochemical staining. Moreover, the messenger ribonucleic acid (mRNA) expressions of NF-κB and ICAM-1 in brain tissues were determined through Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Systolic blood pressure exhibited significant differences among the three groups at 15 d and 19 d of gestational age (p<0.05). At 15 d of gestational age, systolic blood pressure was significantly higher in model group than that of control group (p<0.05). However, it was slightly lower in treatment group than model group (p<0.05). At 19 d of gestational age, systolic blood pressure was significantly higher in model group than control group (p<0.05). However, it decreased remarkably in treatment group when compared with model group (p<0.05). In treatment group, systolic blood pressure at 19 d was significantly lower than that at 15 d (p<0.05). Subsequent Western blotting revealed that the protein expression of B-cell lymphoma-2 (Bcl-2) in brain tissues decreased evidently, whereas the expression of Bcl-2 associated X protein (Bax) increased significantly in model group compared with control group, showing statistically significant differences (p<0.01). The protein expression of Bcl-2 in brain tissues increased significantly, while the expression of Bax declined remarkably in treatment group compared with model group (p<0.01). The number of apoptotic cells in model group and treatment group increased significantly compared with that in control group, with the largest in model group (p<0.05). However, it remarkably declined in treatment group compared with model group (p<0.05). These results suggested that MgSO4 treatment could significantly reduce neuronal apoptosis in PE rats. According to the results of immunohistochemistry, the protein expressions of NF-κB and ICAM-1 in brain tissues were significantly higher in model group and treatment group than those in control group (p<0.05). However, they were significantly lower in treatment group than model group (p<0.05). RT-PCR results manifested that the mRNA expressions of NF-κB and ICAM-1 in brain tissues exhibited evident differences among the three groups (p<0.05). Model group and treatment group showed significantly up-regulated mRNA expressions of NF-κB and ICAM-1 in brain tissues compared with control group (p<0.05). The highest mRNA expression was observed in model group. However, treatment group exhibited remarkably decreased mRNA expressions of NF-κB and ICAM-1 in brain tissues compared with model group (p<0.05). MgSO4 exerts a protective effect on cranial nerves of PE rats by inhibiting the NF-κB/ICAM-1 signaling pathway.

Impact of Xenon on CLIC4 and Bcl-2 Expression in Lipopolysaccharide and Hypoxia-Ischemia-Induced Periventricular White Matter Damage

Background: Premature birth is a significant health care burden. Xenon (Xe) is a general anesthetic with neuroprotective effects. Objectives: Here, we investigate the neuroprotective role of Xe in a lipopolysaccharide (LPS)- and hypoxia-ischemia (HI)-induced white matter damage (WMD) model. Methods: Three-day-old Sprague-Dawley rats were randomly divided into a sham group (group A, n = 24), an LPS + HI group (group B, n = 24), and an LPS + HI + Xe group (group C, n = 72). The onset of Xe inhalation started at 0, 2, and 5 h in subgroups C1, C2, and C3, respectively. Next, we performed TUNEL and hematoxylin and eosin (HE) staining; and examined the expression of CLIC4 and Bcl-2 in brain tissues. Results: HE staining revealed distorted cytoarchitecture, tangled nerve fibers, and pyknosis in group B, while Xe treatment improved these histological alterations in the group C pups. Following LPS and HI insult, the number of apoptotic cells significantly increased in group B at 48 and 72 h (p < 0.05), and Xe significantly alleviated apoptosis (p < 0.001) at 24, 48, and 72 h, respectively. Similarly, CLIC4 mRNA expression was significantly increased in group B (p < 0.05), and Xe produced a marked reduction in CLIC4 mRNA expression in group C subgroups (p < 0.05). Western blotting demonstrated enhanced Bcl-2 expression in group C when compared to group B (p < 0.05). Conclusions: These results demonstrate that LPS and HI successfully induced WMD, and Xe decreased neuronal apoptosis via Bcl-2- and CLIC4-mediated pathways. Moreover, the therapeutic time window of Xe extended for up to 5 h. These findings suggest that Xe can be used as a protective treatment for WMD in premature infants.

Biochemical and Apoptotic Biomarkers of Experimentally Induced Traumatic Brain Injury: In Relation to Time since Death

The present study declares the relationship between the cause of death and postmortem biochemical marker alterations in CSF, serum and plasma. In addition, immunohistochemical and microarchitecture examination of brain tissue of White New Zealand rabbits at different PMI after traumatized death. Thirty adult male White New Zealand rabbits were divided into two main equal groups; the first was physically killed by cervical dislocation and the second through head trauma. Each group was subdivided into three PMIs (zero, 6, and 12hrs PM). CSF was used to detect the levels of K+, Na+, Ca++, albumin While, lactic acid, hypoxanthine, ammonia and uric acid concentrations were measured in plasma. Estimation of HMGB1, IL-1β and TNF-α were assessed in serum. In addition to immunohistochemical observations of Bcl-2 and P53 apoptotic proteins in brain tissue. The results revealed that some of the examined parameters as K+, Na+, albumin, ammonia, hypoxanthine and HMGB1 had the potential role in estimation of PMI at examined time periods in physical and traumatized death. Traumatic death induced severe cerebral hemorrhages and necrosis of cerebral parenchyma than physical death. Immunohistochemical results of P53 and Bcl-2 in brain tissue declared focal positive reactions of some neurons, astrocytes and microglia in different degrees with time since death. It was concluded that biochemical analysis of some body fluids, tissue pathological changes and apoptotic markers are applicable tools for assessing accurate PMI after traumatic brain injury and could have a crucial role in legal medicine

Anti-apoptotic and angiogenic effects of intelectin-1 in rat cerebral ischemia.

Ischemic stroke is an acute life-threatening disease, which causes neurological dysfunction. The formation of new blood vessels around the infarct is vital to the restoration of perfusion and healing of brain tissue. Studies have shown that intelectin-1 (omentin) promotes endothelial cell function and angiogenesis in response to ischemia and inhibits apoptosis in rats with unilateral hind limb surgery. In the present study, we investigated the neuroprotective role of intelectin-1 following focal cerebral ischemia. We specifically assessed the effect of increased expression of intelectin-1 in promoting angiogenesis and reducing apoptosis. The treatment was administered using a lentiviral vector, 7 days prior to surgery. The surgery was performed using the established middle cerebral artery occlusion (MCAO) model in rats, and the outcome was evaluated 7 days after injury. Our results demonstrated a significant reduction in brain infarction volume following LV-intelectin-1 treatment. Additionally, CD34 and capillary density were increased in the cerebral ischemic penumbra. Real-time PCR and Western blot revealed an increased expression of intelectin-1, and phosphorylation of eNOS and AKT with enhanced expression of bcl-2 in brain tissues. These data suggest that the successful delivery of LV-intelectin-1 ameliorated ischemic brain injury. It promoted endothelial cell function and revasc ularization, and inhibited apoptosis in response to ischemia by stimulating the Akt-eNOS signaling pathway.

Mechanism of action of Zhuyu Annao pill in mice with cerebral intrahemorrhage based on TLR4.

To explore the protective effect and possible mechanism of action of Zhuyu Annao pill in mice with intracerebral hemorrhage (ICH). Sixty mice were divided into the control group, hemorrhage group, drug-treated group (after hemorrhage), TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+drug-treated group (after hemorrhage) with 12 in each group. Model of autologous ICH was established in all groups. After drilling and 12h of fasting, models in the control group hemorrhage group and TLR4-knockout hemorrhage group were all drenched with 10mL/kg distilled water by intragastric administration. Models in the drug-treated group and TLR4-knockout hemorrhage+drug-treated group were drenched with 6.25g/kg of Zhuyu Annao pill. All groups were treated for 7d. Longa scoring method was used to measure the neurological defect scores and determine the brain water contents of all groups; ELISA was employed to detect the inflammatory factor interleukin (IL)-6, tumor necrosis factor-α (TNF-α) and IL-1β in brain tissues; and Western blot was applied to test the expression quantities of apoptotic protein Bax and anti-apoptotic protein Bcl-2 in brain tissues. At day 3 and 7, compared with the hemorrhage group, the neurological defect scores of the drug-treated group, TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+drug-treated group decreased significantly (P<0.05). Compared with the hemorrhage group, the brain water contents of the drug-treated group, TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+drug-treated group reduced significantly (P<0.05). Compared with the hemorrhage group, the inflammatory factor IL-6, TNF-α and IL-1β of the drug-treated group, TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+drug-treated group decreased significantly (P<0.05). Compared with the hemorrhage group, the expression of apoptotic protein Bax of the drug-treated group, TLR4-knockout hemorrhage group and TLR4-knockout hemorrhage+drug-treated group decreased significantly and the expression of anti-apoptotic protein Bcl-2 increased significantly (P<0.05). Zhuyu Annao pill can alleviate encephaledema for mice with ICH and reduce inflammatory responses and nerve cell apoptosis. TLR4 can mediate inflammatory injury induced by ICH. Thus, Zhuyu Annao pill can play a protective role for brains by decreasing the expression of TLR4.

Нитрозирующий стресс и апоптоз нейронов са1-зоны гиппокампа в условиях моделирования хронической алкогольной интоксикации: нейропротективные эффекты тиоцетама

Fulfilled researches showed that modeling of chronic alcohol intoxication by daily intragastric ethanol administration to non-pedigreed male white rats (first 10 days – 15% ethanol solution in dose 4 g/kg, next 10 days – 15% ethanol solution in dose 6 g/kg, then 10 days – 25% ethanol solution in dose 4 g/kg) leads to activation of nitrosorbidi stress reactions and initiation of neuro-apoptosis. Thus, as compared with healthy animals group, in rats’ brain after chronic ethanol administration it was revealed significant increase of nitro-tyrosine (nitrosorbidi stress marker) in cytosolic and mitochondrial fractions of brain homogenate against the background of revealing of apoptosis molecular markers in hippocampal CA1 zone neurons (fragmentation of neuron&amp;#180;s nuclei of hippocampal CA1 zone, increase of density and ratio of apoptotic- and destructive-changed cells) as well as deprivation of anti-apoptotic mechanisms (decrease of density of Bcl-2-positive neurons in hippocampal CA1 zone and decrease of Bcl-2 protein concentration in hippocampus). Subsequent 14-day intragastric administration of complex neuro-metabolic cerebro-protector Thiocetam in dose 100 mg/kg to the animals after chronic alcoholization showed significant protective effect on hippocampal CA1 zone neurons. Thus, Thiocetam administration resulted in decrease of nitrotyrosine levels in mitochondrion and in cytosol of brain of animals with chronic alcoholization as compared with untreated animals and with animals receiving Piracetam. Thiocetam administration to experimental animals led to significant decrease of density of apoptotic- and destructive-changed neurons of hippocampal CA1 zone and ratio of apoptotic-changed cells as compared with control group and animals receiving Piracetam. Course of treatment with Thiocetam increases significantly the density of Bcl-2-positive neurons in CA1 zone of hippocampus and significantly increases concentration of Bcl-2 in brain tissue of experimental animals as compared with control group and animals which received Piracetam according the same schedule. The increase of Bcl-2 protein expression in animals received Thiocetam testifies to the activation of anti-apoptotic defense of damaged neurons under the influence of this preparation. We suppose that one of the molecular mechanisms of neuro-protective effect of Thiocetam in chronic intoxication is the breaking of NO-depended mechanisms of neuro-apoptosis.

Changes of neuronic apoptosis, Rho GDP dissociation inhibitor 2 and Bcl-2 in brain tissue of neonatal rats with hypoxic-ischemic brain damage

Objective To investigate the effect of Rho GDP dissociation inhibitor 2 （RhoGDI2） and Bcl-2 in pathogenesis of hypoxic-ischemic brain damage （HIBD）. Methods Neonatal 7-day-old Sprague Dawley rats were randomly divided into sham-operation control group, HIBD 6 h group and HIBD 48 h group （ n = 10 per group）. The apoptosis rate of brain cell was measured by flow cytometer and the expression of RhoGDI2 mRNA and Bcl-2 mRNA were detected by Real-time RT-PCR. Results （ 1 ） The ligated cerebral hemisphere of neonatal rats showed obvious edema at 48 h after hypoxia-ischemia. （ 2 ） Apoptotic cell appeared at 6 h in HIBD group,the apoptosis rate was （ 1.40 ±0. 12） %. The apoptosis rate obviously increased to （ 15.86 ±0.98 ） % at 48 h after HIBD, which showed a significant increase compared to sham-operation control group （ P 〈 0.01 ）. （ 3 ） The expressions of both RhoGDI2 mRNA and Bcl-2 mRNA were 4. 12 ±0. 74 and 2. 55±0. 65 respectively in sham-operation control group. In HIBD group, the expressions of both RhoGDI2 mRNA and Bcl-2 mRNA began to decrease at 6 h after HIBD （3. 19±0. 77,1.96±0. 36） and decreased furthermore at 48 h after HIBD （ 1.04±0. 18,1.06 ±0. 17 ）. The differences of expression levels among three groups were statistically significant （P 〈 0. 01 ）. （4） The expression of RhoGDI2 mRNA positively correlated with the expression of Bcl-2 mRNA （ r = 0. 831, P 〈 0. 05 ）. Conclusion With the emerging of apoptosis after HIBD, the expressions of both RhoGDI2 mRNA and Bcl-2 mRNA are decreased. The imbalance of expression of RhoGDI2 is involved in pathogenesis of HIBD by regulating Bcl-2 expression. Key words: Hypoxic-ischemic brain damage; Rho GDP dissociation inhibitor 2 ; Bcl-2 ; Apoptosis ; Rat, newborn