Investigating the Co-administration of Ghrelin and Nicotine Into the Medial Septum and Its Influencing on Morphine Amnestic Effect.

Introduction: Evidence indicates that medial septum nicotinic receptors regulate cognitive processes. Ghrelin is a gut hormone that regulates energy homeostasis. Ghrelin is also produced in the brain and is involved in cognitive function. This study aims to evaluate the effects of medial septal administration of ghrelin on the amnestic effect of morphine in rats. In addition, the possible relationship between the medial septal ghrelin and acetylcholine nicotinic receptors on the amnestic effect of morphine is evaluated. Methods: The rats were implanted at the medial septum area and were microinjected with ghrelin and nicotinic receptor agents. The step-through type inhibitory avoidance apparatus was used for memory retrieval assessment. Results: The results showed that the administration of morphine after the training phase impaired memory consolidation. Post-training intra-septal injection of the same doses of either ghrelin or nicotine did not change memory performance; however, their co-application with morphine (significant dose: 7.5 mg/kg subcutaneous injection) increased the step-through latency and improved memory consolidation. Moreover, post-training co-application of low doses of the two agonists could not affect morphine-induced memory impairment. Conclusion: These results indicated no interaction between medial septal ghrelin and nicotinic receptors on the amnestic effect of morphine in rats. Highlights: Post-training morphine administration impaired memory performance.Intra-septal injection of ghrelin or nicotine alone did not affect memory performance.Co-application of either ghrelin or nicotine with morphine improved memory.Co-application of the two agents could not affect morphine-induced memory impairment. Plain Language Summary: Morphine abuse has been associated with memory disturbance. Ghrelin is a gastrointestinal hormone known as hunger hormone. It also affects cognitive performance via binding ghrelin receptors in central nervous system. On the other hand, the medial septum nicotinic receptors improve memory-associated behavior. Hence, we hypothesized that septal ghrelin receptors could affect the effect of nicotine on morphine-induced memory deficit. We examined this hypothesis in avoidance memory task. We found that subcutaneous administration of morphine inhibited avoidance memory. The effect of morphine was blocked by intra-medial septum injection of nicotine or ghrelin. However, co-infusion of ghrelin with nicotine into the medial septum area had no effect on morphine amnesia. Overall, the study results suggest no interaction between ghrelin and cholinergic nicotinic receptors in morphine amnesia.

New findings about comparing the effects of antibiotic therapy and phage therapy on memory and hippocampal pyramidal cells in rats.

BACKGROUND: Pseudomonas aeruginosa is a significant cause of infection in burn wounds. Antibiotics are widely used to treat infectious diseases, and alongside their therapeutic benefits, they can damage host cells. Significant side effects, such as nephrotoxicity and neurotoxicity, are observed in 60% of patients treated with colistin. Therefore, using a suitable alternative instead of antibiotics is paramount. This study aimed to investigate the effects of phage therapy and antibiotic therapy on memory function in rats with P. aeruginosa infected burn wounds. METHODS: Adult male rats were divided into three groups: (1) infected without treatment (control), (2) infected and treated with colistin antibiotic (3,000,000 international units/kg/day), and (3) infected and treated with 100 µL of phage suspension (approximately 109 PFU/mL). In all animals, after anesthesia, a third-degree burn was created in the back area. One hour later, treatment was performed for seven consecutive days. Passive avoidance test, novel object recognition test, locomotion activity, hippocampal neuron count, and oxidative stress measurement in blood serum were performed. RESULTS: In antibiotic-treated group memory recall, recognition index, number of healthy neurons in CA1, CA2, and CA3 hippocampus areas and the amounts of MDA, and FRAP significantly decreased compared with the control group. The phage-treated group was not shown any harmful effect on the memory process, number of healthy hippocampal neurons, and showed more positive effects in blood serum examinations compared with the antibiotic group. CONCLUSIONS: Phage therapy could be a safe and effective alternative to antibiotics in the treatment of burn-related infections.

A role for flavonoids in the prevention and/or treatment of cognitive dysfunction, learning, and memory deficits: a review of preclinical and clinical studies.

OBJECTIVE: Natural food substances, due to high rates of antioxidants, antiviral and anti-inflammatory properties, have been proposed to have the potential for the prevention or treatment of cognitive deficits, learning and memory deficits and neuro inflammation. In particular, medicinal plants with rich amounts of beneficial components such as flavonoids are one of the most promising therapeutic candidates for the cognitive deficit and memory loss. Herein, we aimed to review the impact of medicinal plants with focus on flavonoids on cognitive dysfunction, learning and memory loss by considering their signaling pathways. METHODS: We extracted 93 preclinical and clinical studies related to the effects of flavonoids on learning and memory and cognition from published papers between 2000 and 2021 in the MEDLINE/PubMed, Cochrane Library, SCOPUS, and Airiti Library databases. RESULTS: In the preclinical studies, at least there seem to be two main neurological and biological processes in which flavonoids contribute to the improvement and/or prevention of learning, memory deficit and cognitive dysfunction: (1) Regulation of neurotransmission system and (2) Enhancement of neurogenesis, synaptic plasticity and neuronal survival. CONCLUSION: Although useful effects of flavonoids on learning and memory in preclinical investigations have been approved, more clinical trials are required to find out whether flavonoids and/or other ingredients of plants have the potent to prevent or treat neurodegenerative disorders.

Silymarin modulates cadmium-induced oxidative stress in human spermatozoa.

Environmental pollutants such as cadmium can negatively affect sperm parameters and decrease male fertility by inducing oxidative stress. Antioxidants are considered a useful strategy for oxidative stress conditions to neutralize free radicals and strengthen the antioxidant defence system. In this study, the effects of the common application of silymarin, as a natural antioxidant, with cadmium were assessed on human sperm. The washed human sperm samples were divided into five groups: (1) spermatozoa at 0- hour; (2) spermatozoa at 3 h; (3) spermatozoa treated with cadmium (20 µM) for 3 h; (4) spermatozoa treated with silymarin (2 µM) + cadmium (20 µM) for 3 h and (5) spermatozoa treated with silymarin (2 µM) for 3 h. Our results displayed that cadmium reduced sperm motility, viability, plasma membrane integrity and acrosome integrity by increasing malondialdehyde levels and decreasing the total antioxidant capacity and antioxidant enzymes activity. While silymarin attenuated oxidative stress biomarkers in human sperm treated with cadmium, and consequently improved the sperm quality. In summary, cadmium-induced oxidative stress impaired human sperm structures and silymarin with its antioxidant properties compensated for the adverse effects of oxidative stress on human spermatozoa.

Synergistic improvement effect of nicotine-ghrelin co-injection into the anterior ventral tegmental area on morphine-induced amnesia.

In the present study the effect of ghrelin or ghrelin/nicotine injection into the anterior ventral tegmental area (aVTA) on morphine-induced amnesia in passive avoidance learning have been evaluated. Also, the role of the aVTA nicotinic receptors in possible ghrelin-induced effects has been investigated. All animals were bilaterally implanted with chronic cannulas in the aVTA. A step-through type passive avoidance task was used for measurement of memory. We found that post-training subcutaneous (s.c.) injection of morphine (0.5-7.5 mg/kg) dose-dependently reduced the step-through latency, indicating morphine-induced amnesia. Post-training bilateral infusion of ghrelin (0.3, 1.5 and 3 nmol/µl) in a dose-dependent manner reversed amnesia induced by morphine (7.5 mg/kg, s.c.). Furthermore, reversal effect of ghrelin (3 nmol/µl) was blocked by pre-treatment of intra-aVTA administration of mecamylamine (1-3 µg/rat), a nicotinic acetylcholine receptor antagonist. Intra-aVTA administration of the higher dose of mecamylamine (3 µg/rat) into the aVTA by itself decreased the step-through latency and induced amnesia. In addition, post-training intra-aVTA administration of nicotine (0.25, 0.5, 1 µg/rat) which alone cannot affect memory consolidation, decreased significantly the amnesia induced by morphine (7.5 mg/kg, s.c.). Co-treatment of an ineffective dose of ghrelin (0.3 nmol/µl) with an ineffective dose of nicotine (0.25 µg/rat) significantly increased step-through latency of morphine (7.5 mg/kg, s.c.) treated animals, indicating the synergistic effect of the drugs. Taken together, our results suggest that intra-aVTA administration of ghrelin reversed morphine-induced amnesia and that ghrelin interacts synergistically with nicotine to mitigate morphine-induced amnesia.

Impact of aluminium toxicity on vital human sperm parameters-Protective effects of silymarin.

Aluminium is an environmental pollutant which induces oxidative stress, while silymarin is a potent antioxidant. This study was conducted to investigate the possible protective effects of silymarin on adverse effects of aluminium chloride on vital sperm parameters as well as its effects on oxidative stress markers in human spermatozoa. Human spermatozoa were divided into 5 groups as follows: (a) spermatozoa at 0 hr; (b) spermatozoa at 180 min (control); (c) spermatozoa treated with aluminium chloride; (d) spermatozoa treated with silymarin + aluminium chloride; and (e) spermatozoa treated with silymarin. The sperm samples were used to assess sperm vital parameters such as acrosome and plasma membrane integrity, viability, mitochondrial membrane potential (MMP) and motility as well as sperm malondialdehyde (MDA) level and the total antioxidant capacity. The percentage of acrosome and plasma membrane integrity, viability, MMP, motility and the total antioxidant capacity of spermatozoa treated with aluminium chloride significantly decreased compared with control group, while the level of MDA significantly increased compared with the control group. In the silymarin + aluminium chloride group, silymarin could significantly compensate the adverse effects of aluminium chloride on these parameters. Administration of silymarin alone significantly increased the percentage of acrosome and plasma membrane integrity, viability, motility and total antioxidant capacity, while significantly reduced MDA levels compared with the control group. Aluminium chloride by inducing oxidative stress exerts disastrous effects on the vital parameters of human spermatozoa and silymarin, as a potent antioxidant, could reverse the effects of aluminium chloride on these parameters.

Ghrelin modulates morphine-nicotine interaction in avoidance memory: Involvement of CA1 nicotinic receptors.

Ghrelin is a stomach-derived hormone which regulates appetite and energy balance in the body. Recent studies show that ghrelin has been linked to the learning and memory process. Ghrelin also modulates reward properties of addictive drugs. However, the involvement of ghrelin in cognitive effects of addictive drugs has not been examined yet. The goal of present study is to examine the effect of intra-CA1 administration of ghrelin on morphine response for avoidance task alone or in combination with nicotine. Here, we also investigated the role of hippocampal nicotinic cholinergic receptors in possible interaction of the drugs in adult male Wistar rats. Results showed that subcutaneous administration of morphine immediately after training impaired memory in the test day and induced amnesia, while intra-CA1 pre-injection of ghrelin prevented amnesic effect of morphine and improved memory. Also, systemic administration of nicotine five min prior to morphine administration dose-dependently inhibited morphine-induced amnesia. The results showed that intra-CA1 injection of an ineffective dose of ghrelin (0.03 nmol/µl) potentiated the nicotine (0.2 mg/kg, s.c.) response on amnesia induced by morphine. This stimulatory effect was inhibited by mechamylamine, a non-competitive nicotinic receptor antagonist. Moreover, post-training administration of drugs (ghrelin, nicotine and mecamylamine) alone had no effect on memory consolidation. In conclusion, present study suggests the significant role of ghrelin in morphine-related memory and its interactive effect with nicotine in avoidance task via CA1 nicotinic receptors.

Myricetin protects hippocampal CA3 pyramidal neurons and improves learning and memory impairments in rats with Alzheimer's disease.

There is currently no treatment for effectively slowing the progression of Alzheimer's disease, so early prevention is very important. Numerous studies have shown that flavonoids can improve memory impairment. The present study investigated the effects of myricetin, a member of the flavonoids, on intracerebroventricular streptozotocin induced neuronal loss and memory impairment in rat models of Alzheimer's disease. Myricetin at 5 or 10 mg/kg was intraperitoneally injected into rats over 21 days. Control rats were treated with 10 mL/kg saline. Behavioral test (the shuttle box test) was performed on day 22 to examine learning and memory in rats. Immediately after that, hematoxylin-eosin staining was performed to observe the morphological change in hippocampal CA3 pyramidal neurons. Myricetin greatly increased the number of hippocampal CA3 pyramidal neurons and improved learning and memory impairments in rats with Alzheimer's disease. These findings suggest that myricetin is beneficial for treatment of Alzheimer's disease.

Proteome Analysis of Rat Hippocampus Following Morphine-induced Amnesia and State-dependent Learning.

Morphine's effects on learning and memory processes are well known to depend on synaptic plasticity in the hippocampus. Whereas the role of the hippocampus in morphine-induced amnesia and state-dependent learning is established, the biochemical and molecular mechanisms underlying these processes are poorly understood. The present study intended to investigate whether administration of morphine can change the expression level of rat hippocampal proteins during learning of a passive avoidance task. A step-through type passive avoidance task was used for the assessment of memory retention. To identify the complex pattern of protein expression induced by morphine, we compared rat hippocampal proteome either in morphine-induced amnesia or in state-dependent learning by two-dimensional gel electerophoresis and combined mass spectrometry (MS and MS/MS). Post-training administration of morphine decreased step-through latency. Pre-test administration of morphine induced state-dependent retrieval of the memory acquired under post-training morphine influence. In the hippocampus, a total of 18 proteins were identified whose MASCOT (Modular Approach to Software Construction Operation and Test) scores were inside 95% confidence level. Of these, five hippocampal proteins altered in morphine-induced amnesia and ten proteins were found to change in the hippocampus of animals that had received post-training and pre-test morphine. These proteins show known functions in cytoskeletal architecture, cell metabolism, neurotransmitter secretion and neuroprotection. The findings indicate that the effect of morphine on memory formation in passive avoidance learning has a morphological correlate on the hippocampal proteome level. In addition, our proteomicscreensuggests that morphine induces memory impairment and state-dependent learning through modulating neuronal plasticity.

Glucocorticoid-induced impairment of long-term memory retrieval in female rats: influences of estrous cycle and estrogen.

Using an inhibitory avoidance (IA) task, the effects of glucocorticoids on memory retrieval in intact and ovariectomized (OVX) female rats were investigated. Young adult female rats were trained in a one trial IA task (1-mA, 3-s footshock). The latency to reenter the dark compartment of the apparatus was recorded in the retention test performed 48h after training. Pre-retrieval injection of corticosterone (CORT, 1, 3, and 10mg/kg) to OVX rats impaired memory retrieval at all doses tested. Similar administration of CORT (3mg/kg) in intact female rats impaired memory retrieval in the estrus phase (when endogenous plasma levels of estrogen are low) but not in the proestrus phase (when endogenous levels of estrogen are high). Concurrent administration of CORT (3mg/kg) and 17-ß-estradiol (15µg/kg) in both proestrus and estrous phases impaired memory retrieval. Our findings indicate that the effects of corticosterone on memory retrieval are modulated by the estrous cycle and 17-ß-estradiol.

Modulation of morphine state-dependent learning by muscarinic cholinergic receptors of the ventral tegmental area.

In the present study, the effects of bilateral intra-ventral tegmental area (intra-VTA) injections of an anticholinesterase, physostigmine and/or muscarinic acetylcholine receptor antagonist, atropine on memory retention and morphine state-dependent learning were examined in adult male Wistar rats. As a model of learning, a step-through passive avoidance task was used. Post-training subcutaneous administration of morphine (0.5, 2.5 and 5 mg/kg) dose-dependently impaired memory retrieval on the test day. Pre-test administration of morphine (2.5 and 5 mg/kg) induced state-dependent retrieval of the memory acquired under post-training morphine influence. Pre-test intra-VTA microinjection of physostigmine (0.5, 1 and 2 microg/rat) or atropine (1, 2 and 3 microg/rat) alone cannot affect memory retention. Interestingly, pre-test intra-VTA administration of physostigmine (1 and 2 microg/rat) reversed post-training morphine (5 mg/kg, s.c.)-induced retrieval impairment. Furthermore, pre-test intra-VTA microinjection of physostigmine (1 and 2 mug/rat) with an ineffective dose of morphine (0.5 mg/kg), synergistically improved memory performance impaired by post-training morphine. On the other hand, pre-test intra-VTA microinjection of atropine (2 and 3 microg/rat) 5 min before the administration of morphine (5 mg/kg, s.c.) dose-dependently inhibited morphine state-dependent memory. Pre-test atropine microinjection also reversed the influence of physostigmine on morphine response. It may be concluded that the muscarinic acetylcholine receptors of the VTA play an important role in morphine-induced recovery of memory, on the test day.

Nicotinic acetylcholine receptors of the ventral tegmental area are involved in mediating morphine-state-dependent learning.

In the present study, the possible role of nicotinic acetylcholine (nACh) receptors of the ventral tegmental area (VTA) on morphine-state-dependent learning was studied in adult male Wistar rats. As a model of memory, a step-through type passive avoidance task was used. All animals were bilaterally implanted with chronic cannulae in the VTA, trained using a 1mA foot shock, and tested 24h after training to measure step-through latency. Post-training subcutaneous (s.c.) injection of morphine (0.5-5mg/kg) dose-dependently reduced the step-through latency, showing morphine-induced amnesia. Amnesia induced by post-training morphine was significantly reversed by pre-test administration of morphine (2.5-5mg/kg, s.c.) and induced morphine-state-dependent learning. Pre-test injection of nicotine (0.25-1microg/rat) into the VTA plus an ineffective dose of morphine (0.5mg/kg) significantly restored the memory retrieval. It should be noted that pre-test intra-VTA injection of the same doses of nicotine (0.25-1microg/rat) alone cannot affect memory retention. Furthermore, pre-test intra-VTA injection of the nicotinic acetylcholine receptor antagonist, mecamylamine (1-3microg/rat) 5min before the administration of morphine (5mg/kg, s.c.) dose-dependently inhibited morphine-state-dependent learning. Pre-test injection of the higher dose of mecamylamine (3microg/rat) into the VTA by itself decreased the step-through latency and induced amnesia. On the other hand, mecamylamine (0.5 and 1microg/rat, intra-VTA) reversed the effect of nicotine on morphine response. The results indicate that nACh receptors in the VTA participate in the modulation of morphine-induced recovery of memory, on the test day.