Neuroprotective Therapeutic Potential of microRNA-149-5p against Murine Ischemic Stroke.

Ischemic stroke resulting from blockade of brain vessels lacks effective treatments, prompting exploration for potential therapies. Among promising candidates, microRNA-149 (miR-149) has been investigated for its role in alleviating oxidative stress, inflammation, and neurodegeneration associated with ischemic conditions. To evaluate its therapeutic effect, male Wistar rats were categorized into five groups, each consisting of 27 rats: sham, MCAO, lentiviral control, lentiviral miR-149, and miR149-5p mimic. Treatments were microinjected intracerebroventricularly (ICV) (right side), and ischemia was induced using middle cerebral artery occlusion (MCAO) procedure. Post-MCAO, neurological function, histopathological changes, blood-brain barrier (BBB) permeability, cerebral edema, and mRNA levels of Fas ligand (Faslg) and glutamate ionotropic NMDA receptor 1 (GRIN1) were assessed, alongside biochemical assays. MiR-149 administration improved neurological function, reduced brain damage, preserved BBB integrity, and attenuated cerebral edema. Upregulation of miR149-5p decreased Faslg and GRIN1 expression in ischemic brain regions. MiR-149 also reduced oxidative stress, enhanced antioxidant activity, decreased caspase-1 and - 3 activity, and modulated inflammatory factors in ischemic brain regions. Moreover, DNA fragmentation as an index of cell death decreased following miR-149 treatment. In conclusion, the study underscores miR-149 potential as a neuroprotective agent against ischemic stroke, showcasing its efficacy in modulating various mechanisms and supporting its candidacy as a promising therapeutic target for innovative strategies in stroke treatment.

Expression of Na+/Ca2+ exchangers was enhanced following pre-treatment of olive leaf extract and olive oil in animal model of ischemic stroke.

BACKGROUND: Neuroprotective role of olive and its natural products can introduce them as alternative candidates for the management of neurodegenerative diseases including stroke. The present study was designed to evaluate whether pretreatment of olive oil and leaf extract can attenuate the most important destructive processes in cerebral ischemia called excitotoxicity. MATERIAL AND METHODS: The male rats were categorized into control, virgin olive oil (OVV), MCAO, MCAO + OVV (with doses of 0.25, 0.50 and 0.75 ml/kg as treatment groups), olive leaf extract, MCAO + olive leaf extract (with doses 50, 75 and 100 mg/kg as treatment groups) groups. Rats of treatment groups received gastric gavage with olive oil or leaf extract for 30 consecutive days. After pretreatment, the intraluminal filament technique was used to block middle cerebral artery (MCA) transiently. Neurological deficits, infarct volume and expression of Na+/Ca2+ exchangers (NCX1, NCX2 and NCX3) proteins were measured. RESULTS: The results revealed that olive oil at doses of 0.50 and 0.75 ml/kg reduced the infarction and neurological score and upregulated NCXs expression in rat brain. In addition, olive leaf extract at doses of 75 and 100 mg/kg attenuated the infarction and neurological score and enhanced NCXs expression in rat brain. CONCLUSION: These findings support the view that olive oil and leaf extract play the neuroprotective role in cerebral ischemia due to the upregulation of NCXs protein expression.

Excessive audio-visual stimulation leads to impaired social behaviour with an effect on amygdala: Early life excessive exposure to digital devices in male rats.

Today, the effect of extreme early-life exposure to digital devices is suggested as a risk factor for neurodevelopmental disorders. However, the multitude of factors that influence brain development with subsequent behavioural abnormalities have not been fully elucidated. Herein, we simulated extreme early-life exposure to digital devices in rats by audio and visual stimulation and investigated its effects on autism-related behaviours and brain structural alteration. Male rat pups were exposed to excessive audio-visual stimulation (EAVS) from PND (post-natal day) 12 to PND 35, with and without maternal separation (MS). Autism-related behaviours including abnormal sociability, stereotype behaviours, anxiety and locomotor dysfunction were tested at PND 42. Brain structural alternation was examined by considering the amygdala, mPFC (medial prefrontal cortex) and hippocampal regions while performing 3D quantitative stereological analysis. We found that EAVS led to social behaviour deficit and higher locomotion in rats, which were associated with increases in the number of neurons and volume of the amygdala. We also showed that MS did not exaggerate the effect of extreme sensory stimulation on behaviour and the structure of the brain. This study proposed EAVS in rats as an animal model of early exposure to digital devices for investigating possible neurobiological alternations underlying autistic-like behaviours with an emphasis on the amygdala area.

Reduced expression of apoptotic proteins in the ischemic rat brain following Sertoli cell transplantation.

Sertoli cells (SCs) may be a new candidate to decrease ischemic damage due to their ability to secrete factors that actively protect neurons and inhibit uncontrollable immune responses. Pre­treatment with these cells was considered in the current study. SCs were injected into the right striatum in rats using the stereotaxic technique. Ten days after injection, middle cerebral artery occlusion surgery was performed. Following these procedures, neurological deficit scores, brain edema, blood­brain barrier integrity, infarct volume, and the expression of apoptotic factors in the cortex, striatum, and piriform cortex­amygdala were evaluated. Analysis showed that behavioral deficits, infarct volume, blood­brain barrier permeability, and edema in the striatal area in the allograft group demonstrated a significant decrease compared to the control group. Additionally, analysis of the expression of caspase­3 and Bcl­2 proteins in the striatum indicated a remarkable reduction and increase, respectively, in the allograft group compared to the control group. According to the obtained results, one possible mechanism for the neuroprotection induced by SCs in an ischemic brain is the reduction of apoptotic factors.

Antioxidant and anti-apoptotic effects of cannabidiol in model of ischemic stroke in rats.

One of the main non-psychoactive phytocannabinoids of cannabis is cannabidiol (CBD), which has attracted much attention for its neuroprotective roles. The present study was designed to assess whether pretreatment of CBD can attenuate two of the destructive processes of cerebral ischemia, including oxidative stress and cell death. The male rats were randomly divided into 6 main groups (control, MCAO, vehicle, and CBD-treated groups). Using stereotaxic surgery, a cannula was inserted into the right lateral ventricle of the rat brain. CBD was injected at doses of 50, 100 and 200 ng/rat for five consecutive days. After pretreatment, middle cerebral artery (MCA) was blocked for 60 min using the intraluminal filament technique. 24 h after reperfusion, each main group was considered for measurement of infarct volume, superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), p53 gene expression, pathological alterations, and expression of Bax, Bcl-2, cytochrome C, and caspase-3 proteins. The results revealed that CBD at dose of 100 ng/rat reduced the infarction volume and MDA level in cortical and striatal areas of rat brain compared with vehicle group. In addition, the CBD at dose of 100 ng/rat elevated the activity of SOD enzyme in cortex and striatum. The increase in the activity of CAT was also seen at dose of 100 ng/rat in cortex. Furthermore, the Bcl-2/Bax ratio was significantly diminished by the dose of 100 ng/rat CBD in cortex. Moreover, a decrease in expression of cytosolic cytochrome C was observed by CBD at doses of 100 and 200 ng/rat in cortex. CBD at doses 100 and 200 ng/rat also reduced the expression of caspase-3 in cortical and striatal areas, respectively. P53 was downregulated following administration of CBD at dose of 100 ng/rat. Moreover, histological analysis showed the decrease in the percentage of pyknotic neurons in 100 and 200 ng/rat CBD-received groups. CBD played the anti-apoptosis and anti-oxidant roles in cerebral ischemia by affecting the pathways of intrinsic apoptosis, endogenous antioxidant enzymes, and lipid peroxidation.

Effect of Sertoli Cell Transplant and Rapamycin Pretreatment on Middle Cerebral Artery Occlusion-Induced Brain Ischemia in a Rat Model.

OBJECTIVES: Stroke exacts a heavy toll on death and disability worldwide. In animal studies, cell transplant has shown a positive effect by inducing neurogenesis, angiogenesis, and modulating inflammation. Cell transplant therapy could provide researchers with new strategies for treating stroke. The mechanistic target of rapamycin is a central signaling pathway for coordination and control; the administration of rapamycin, a key modulator of this pathway, could be a new therapeutic approach in neurological disorders. MATERIALS AND METHODS: Adult rats were grouped into 5 main groups: control, sham, rapamycin receiving, Sertoli cell receiving, and rapamycin plus Sertoli cell receiving groups. Sertoli cells were taken from another rat tissue and injected into the right striatum region. After 5 days, ischemic induction was performed, and rapamycin injection (300 mg/kg) was performed 1 hour before surgery. After 24 hours, some regions of the brain, including the cortex, striatum, and piriform cortex-amygdala, were isolated for evaluation. RESULTS: Our results showed that infarct volume, brain edema, and blood-brain barrier permeability assessments were significantly reduced in some areas of the brain in rats that received rapamycin plus Sertoli cells compared with results shown in the control group. CONCLUSIONS: Pretreatment with Sertoli cell transplant plus rapamycin injection may enhance neural survival during ischemia through increased glial cell-derived neurotrophic factor and vascular endothelial growth factor, inhibiting the mechanistic target of rapamycin pathway and increasing autophagy performance.

Hypericin Ameliorates Maternal Separation-Induced Cognitive Deficits and Hippocampal Inflammation in Rats.

BACKGROUND AND OBJECTIVE: Maternal separation as an epigenetic agent provokes a severe change in the brain, such as inflammation response, which is a key risk factor for the progression of autism spectrum disorders (ASD). This study evaluated the preventive effect of hypericin on maternal separation-induced cognitive deficits and hippocampal inflammation. METHODS: Here, we reported that pups are subjected to maternal separations for 1 h per day from postnatal days (PND) 1-9 displayed apparent memory impairment in young rats (postnatal day 34) compared to controls group. Furthermore, maternal separation significantly increased inflammation factors in the hippocampus area. Anti-inflammation constituent shed light on treating ASD. RESULTS: In this study, we found that treatment with hypericin (10 and 50 mg/kg) significantly suppresses expression of hippocampal interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in the maternal separation rat model. Also, we found that hypericin prevented the decrease of hippocampal dopamine levels in the offspring of maternal separation rats. CONCLUSION: The data indicated that hypericin may play a neuroprotective role in hippocampal cell and ameliorates dysfunctions in memory and level of inflammation factor in this autism model. Thus, hypericin could be used as an intervention for treating ASD.

Allograft of Sertoli Cell Transplantation in Combination with Memantine Alleviates Ischemia-Induced Tissue Damages in An Animal Model of Rat.

OBJECTIVE: Brain ischemia is the most common disease in the world caused by the disruption of the blood supply of brain tissue. Cell therapy is one of the new and effective strategies used for the prevention of brain damages. Sertoli cells (SCs) can hide from the host immune system and secrete trophic factors. So, these cells have attracted the attention of researchers as a therapeutic option for the treatment of neurodegenerative diseases. Also, memantine, as a reducer of glutamate and intracellular calcium, is a suitable candidate for the treatment of cerebral ischemia. The principal target of this research was to examine the effect of SC transplantation along with memantine on ischemic injuries. MATERIALS AND METHODS: In this experimental research, male rats were classified into five groups: sham, control, SC transplant recipient, memantine-treated, and SCs- and memantine-treated groups. SCs were taken from another rat tissue and injected into the right striatum region. A week after stereotaxic surgery and SCs transplantation, memantine was injected. Administered doses were 1 mg/kg and 20 mg/kg at a 12-hour interval. One hour after the final injection, the surgical procedures for the induction of cerebral ischemia were performed. After 24 hours, some regions of the brain including the cortex, striatum, and Piriform cortex-amygdala (Pir-Amy) were isolated for the evaluation of neurological deficits, infarction volume, blood-brain barrier (BBB) permeability, and cerebral edema. RESULTS: This study shows that a combination of SCs and memantine caused a significant decrease in neurological defects, infarction volume, the permeability of the blood-brain barrier, and edema in comparison with the control group. CONCLUSION: Probably, memantine and SCs transplantation reduce the damage of cerebral ischemia, through the secretion of growth factors, anti-inflammatory cytokines, and antioxidant factors.

GCH1 (rs841) polymorphism in the nitric oxide-forming pathway has protective effects on obstructive sleep apnea.

Several studies have recently investigated the contribution of genetic factors in obstructive sleep apnea (OSA). Patients with OSA suffer from a reduction in nitric oxide (NO) serum level. This study investigated rs841, A930G p22phox, and rs1799983 polymorphisms in three critical genes involved in NO formation. A total of 94 patients with OSA and 100 healthy controls were enrolled into the study. Results showed there was no association between rs841, A930G p22phox and rs1799983 polymorphism and the risk of OSA (P = 0.51, P = 0.4 and P = 0.33, respectively). Moreover, rs841 GA genotype had a reverse relationship with the severity of OSA (P = 0.005). On the other hand, rs841 GA and A930G p22phox AA genotypes had a protective effect on daytime sleepiness in OSA patients (P = 0.01and P = 0.02, respectively). Additionally, the combination of rs841 and A930G p22phox (AG/AG and AG/AA) genotypes was significantly associated with a reduction in daytime sleepiness in OSA patients (P = 0.03 and P = 0.03, respectively). According to the results of our study, GA genotype of rs841 and GA/AA genotypes of A930G p22phox polymorphisms significantly reduced the severity of the problem and daytime sleepiness in OSA patients.

Differential impact of treadmill training on stroke-induced neurological disorders.

OBJECTIVE: Physical exercise contributes to improving stability against nerve injury caused by ischaemic stroke. Here we aimed to preliminarily investigate the effects of continuous endurance training (CET) and high-intensity interval training (HIT) on stroke-associated anxiety, locomotion, neurological assessments and P70S6 Kinase (P70S6K) activation as well. To do this, rats were trained according to HIT and CET protocols for 2 months prior to being subject to middle cerebral artery occlusion surgery. METHODS: Twenty-four hours later behavioural examination was performed by elevated plus maze (EPM) testing, open field and neurological scoring followed by cortical and hippocampal P70S6Ks immunoblotting. RESULTS: According to the obtained data pre-ischaemic HIT and CET similarly improved neurological performance, anxiety levels and locomotion in EPM and open field tests following ischaemic stroke while there was a remarkable rise in hippocampal and cortical P70S6K activation in the HIT group compared to the CET counterparts. CONCLUSION: Behavioral and molecular data suggest that interval training is more beneficial rather than CET, but the distinct mechanisms of CET and HIT on memory are still topics to be discovered.

Investigating effects of atmospheric-pressure plasma on the process of wound healing.

Cold atmospheric-pressure plasma jets (APPJ) have excellent applications in biomedicine. Advantages of APPJ include lack of need for vacuum systems, capability of operation for a long time, and safe to be directly touched by living tissues such as a human body. In this study, an APPJ was generated by a dielectric barrier and applied for the treatment of chemical wounds. This APPJ worked with argon and was driven by high-voltage pulses. This paper compares the spontaneous healing of wounds and a stimulated healing using daily APPJ treatment. Biological data, such as hematological, biochemical, and histological parameters, were remarked. The mortality and morbidity of the untreated samples were reported after 20 days in comparison with the plasma-treated samples, which were alive after these days. Experimental results demonstrated that an increase in the oxidative stress could result in the decreased destruction of lesions by controlling the infection growth. These results were related to the presence of reactive oxygen species and reactive nitrogen species in the plasma volume, which were detected by optical emission spectroscopy.