While these associations are valid, there's an equally important yet lesser-known dimension to their role — their significant impact on cognitive well-being. Before exploring the brain-boosting benefits, let's briefly recap what electrolytes are. Electrolytes are minerals that carry an electric charge when dissolved in bodily fluids like blood and cells.

These ions or charged particles play a crucial role in various physiological processes within the human body and other organisms. Electrolytes are essential for many important bodily functions , including maintaining proper fluid balance, nerve function, muscle contractions, and the functioning of many cellular processes.

They help transmit electrical signals in nerve cells, regulate muscle contractions including the heartbeat , and assist in maintaining the pH balance of bodily fluids.

Potassium plays a substantial role in cognitive function. This critical electrolyte helps maintain the electrical activity of neurons, allowing them to communicate effectively.

Here's how potassium benefits cognitive health:. Signal Transmission: Potassium is crucial for the release and action of neurotransmitters , which are chemicals that transmit signals between nerve cells neurons.

Neurons communicate with each other by releasing neurotransmitters into synapses — the tiny gaps between neurons. These neurotransmitters carry signals from one neuron to the next, allowing for the transmission of information throughout the brain.

Memory and Learning: The effective release and action of neurotransmitters are crucial for processes like memory formation and learning.

For example, when you learn something new, neurotransmitters are involved in strengthening connections synapses between neurons, making it easier for you to recall and apply that knowledge later.

Oxygen and Nutrient Supply: Potassium helps regulate blood flow to the brain , ensuring that brain cells receive an adequate supply of oxygen and nutrients.

The brain is highly metabolically active and requires a continuous delivery of oxygen and glucose a nutrient to function optimally. Potassium contributes to the dilation widening of blood vessels in the brain , facilitating the delivery of these essential resources.

Enhanced Cognitive Performance: When brain cells receive sufficient oxygen and nutrients, they can perform at their best.

This can lead to enhanced cognitive performance, including improved focus, attention, and problem-solving abilities. Generating Action Potentials: Nerve impulses, also known as action potentials, are the electrical signals that neurons use to transmit information rapidly.

Potassium plays a crucial role in generating and maintaining these action potentials. Thinking and Movement: Nerve impulses are responsible for various cognitive functions, such as thinking, decision-making, and voluntary muscle movements. When you think or decide to move a part of your body, it's because neurons are firing action potentials, and potassium is involved in these electrical processes.

A deficiency in potassium hypokalemia can lead to symptoms like fatigue, muscle weakness, and impaired cognitive function, including difficulty concentrating and memory problems. Magnesium is another electrolyte that is vital for brain health.

It's involved in over biochemical reactions in the body, many of which are linked to cognitive function. Here are some of the ways magnesium supports your brain:.

Adaptive Learning: Magnesium is crucial for neuroplasticity , which is the brain's ability to adapt, reorganize, and form new neural connections. One study found that magnesium can influence learning and memory in rodents and improve cognitive performance in brain injury scenarios.

Stress Response Regulation: Magnesium plays a role in regulating the body's stress response. Chronic stress can have detrimental effects on the brain, leading to cognitive impairments and emotional disturbances. Magnesium helps mitigate these effects by acting as a natural relaxant. It counteracts the release of stress hormones like cortisol, promoting a sense of calm and reducing the negative impact of stress on the brain.

Gamma-Aminobutyric Acid GABA Stimulant: Magnesium helps alleviate stress by binding to and activating GABA receptors in the brain. GABA , a key neurotransmitter, helps slow down brain activity, promoting relaxation. Low GABA levels can lead to heightened stress, manifesting as overwhelming feelings, disorganization, excessive worrying, and insomnia due to racing thoughts.

This deficiency is associated with various stress-related disorders. Emotional Well-Being: Both animal and human studies suggest that magnesium has a positive impact on mood.

In one animal study , mice with low magnesium levels displayed higher aggression and restlessness, along with increased adrenaline. Human research showed that taking mg of magnesium daily for 90 days under mental pressure improved physiological regulation and mental balance, helping prevent symptoms like restlessness, irritability, and sleeplessness associated with magnesium deficiency.

A deficiency in magnesium hypomagnesemia can result in other symptoms such as brain fog, anxiety, and memory issues. Homogenization was carried out in 0. The homogenate was then centrifuged at 10, rpm 6, xg for 10 min, and the clear supernatant obtained was used for estimating biochemical parameters Hota et al.

Briefly, cell lysate for GSSG and total glutathione was prepared using a specific cell lysis buffer. Deproteination was performed using metaphosphoric acid MPA reagent. The OD was taken at nm at 0 min and again at 10 min.

Briefly, 1. After that, incubation was done for 15 min at 37°C in the dark, and fluorescence was measured at nm excitation and nm emission. The values obtained were transformed to fluorescent units per milligram of protein by calculating the protein found in 25 µL of the respective sample from a standard curve Hota et al.

The measurement of lipid peroxidation was done spectrophotometrically by calculating MDA formed as a product, as designated by Utley et al. A respective molecule of malondialdehyde reacts with two molecules of thiobarbituric acid TBA to produce a colored MDA—TBA complex that can be further measured spectrophotometrically at nm Hota et al.

LDH activity in the hippocampus tissue was estimated using the LDH assay kit from RANDOX RANDOX Laboratory Ltd. The assay was performed as per the protocol suggested by the manufacturer with minor modifications. Briefly, the assay was carried out by adding 10 μL of sample to μL of the reconstituted reagent containing pyruvate and NADH in phosphate buffer.

Phosphate buffer was added as a blank. The absorbance was then taken at nm in an enzyme-linked immunosorbent assay ELISA reader after 30 s that was considered at 0 min and then after 1, 2, and 3 min from the initial reading. The sample and standard were prepared as per the protocol, and the plate was thoroughly mixed and incubated for 10 min at room temperature.

The OD was taken at nm on a Synergy H4 hybrid reader BioTek. Briefly, the glutamate enzyme mix and glutamate developer were solubilized; standards and samples were prepared as per the instructions. The plate was incubated at 37°C for 30 min protected from light. The OD of the plate was taken at nm.

For neurodegeneration study, the brain sections were stained with Fluoro-Jade B, a polyanionic fluorescence derivative that selectively binds to degenerating neurons. Staining was carried out as per the standard protocol Kushwah et al. The samples were viewed under the fluorescence microscope Olympus, Japan by means of the FITC filter.

A positive green fluorescence signal designates degenerative neurons. Morphological alterations were observed by CV staining. CV is commonly utilized to stain Nissl material in the cytoplasm of neurons in PFA- or formalin-fixed tissue.

This was performed as per our previously published protocol Kushwah et al. MWM data were analyzed using the ANY-maze software. For the behavioral study, 7—10 rats in each group were taken. Histology data IHC, Fluoro-Jade B—positive cells, and pyknotic cells were investigated via ImageJ software.

Briefly, images were taken of the whole hippocampus from six different rats, and six random sections were taken from each rat and mounted on two slides thus, each slide comprises three sections from each experimental group.

For neutral investigation, several images of diverse regions of the hippocampus were taken from each section at ×20 magnification. Out of three sections from each slide, nine random images were occupied, and thus, a total of 18 random images were taken from two slides, i.

Densitometry analysis was performed for all immunoblots. Data for all experimentations were articulated as mean ± SEM. Statistical investigations were performed by GraphPad Prism 5 throughout the study unless specified otherwise. The study has been conducted to explore the effect of GBE on HH-induced damage at behavioral and molecular levels.

Further mechanisms of action were also explored. GBE comprises different Ginkgo flavone glycosides, and we estimated quercetin using HPTLC Supplementary Figures 1A,B. The analysis showed quercetin was found as a major constituent at Rf 0. The content of quercetin in GBE was found as HH-mediated memory impairment has been well studied in various studies Shukitt-Hale et al.

A similar result was observed in the present study which shows a significant increase in latency to reach the platform and path length after 14 days of HH exposure Figures 2A,B. FIGURE 2. GBE improves the spatial memory performance of rats in the MWM.

The effect of GBE on memory was studied using the MWM spatial memory test. Treatment of rats with GBE showed protective effect on HH-induced memory impairment as evident from the decreased latency and path length B and increased time spent and number of entries in the target quadrant D as compared to the HH group.

One-way ANOVA with the Bonferroni test was used to analyze the data. Data are represented as mean ± SEM. Oxidative stress is known to be one of the major causes of HH-induced cognitive deficit. Hence, we further explored the effect of GBE on oxidative stress markers and apoptosis.

Free radical generation during oxidative stress was measured as ROS level estimation by the DCFDA method. FIGURE 3. GBE prevents HH-induced oxidative damage. Representative bar graphs of oxidative stress markers such as ROS level A , lipid peroxidation MDA B , GSH C , GSSG D , and TAC E.

GSH was decreased after 14 days of HH exposure, and GBE treatment significantly reverted these effects. HH increased the level of GSSG, which was further reduced by GBE treatment. The results are expressed as mean ± SEM. Furthermore, we also studied lipid peroxidation via estimating the MDA level.

Antioxidant status was also studied via measuring the intracellular GSH level, which is an important component of the cellular antioxidant defense system and expressed in cells in two different states, i.

Neurodegeneration or cell death was studied at different levels and approaches, i. Initially, cell death was studied biochemically by estimating lactate dehydrogenase LDH levels in different groups.

GBE administration reduced the LDH level to a significant level and hence reduced cell death Figure 4A. FIGURE 4. GBE prevents LDH leakage and ameliorates neuronal damage and morphological alterations during HH exposure. Bar graph of LDH A that shows decreased LDH release after GBE treatment which was increased in HH exposure.

HH leads to neurodegeneration as evident from the increased number of Fluoro-Jade B—positive cells in the hippocampus. Images and bar graph show an increase in Fluoro-Jade B—positive cells in 14 days of HH exposure as compared to the GBE-treated group which significantly reduced the Fluoro-Jade B—positive cells B.

Cresyl violet staining was done to assess pyknotic cells which were higher in the HH group, whereas GBE significantly decreased the pyknotic cell count when compared to the HH group C.

Black arrows indicate pyknotic cells. The scale bar represents 25 µm. Apoptosis is one of the major processes leading to neurodegeneration during HH exposure. Further activated caspase-3 expression, an apoptotic marker, was examined. Immunohistochemistry showed increased apoptosis in the hippocampus at 14 days of HH exposure as evident from the increased number of activated caspase-3—positive cells in the HH group.

FIGURE 5. GBE reduces HH-induced apoptosis in the hippocampus. Images show the caspase-3—positive cells in different groups A. Graphical representation indicates the increased number of caspase-3—positive cells in 14 days of HH exposure, whereas GBE significantly reduced the caspase-3—positive cells when compared to the HH group B.

Black arrows indicate activated caspase-3—positive cells. Immunoblot results also indicate reduced expression of active caspase-3 in groups treated with GBE when compared to the HH group C.

Previously, we have shown increased activity and expression of SK2 channels as one of the causes for HH-induced deterioration Kushwah et al. The expression of SK2 channels significantly reduced in groups administered GBE Figures 6A,B when compared to a group exposed to HH alone for 14 days.

Also, after estimating the glutamate level in different groups, it was observed that 14 days of HH exposure caused glutamate excitotoxicity as evident from the increased glutamate level Figure 6A.

On the contrary, the glutamate level significantly reduced in the GBE-treated group in comparison with the HH alone group. FIGURE 6. GBE reduces expression of the SK2 channel. Immunoblot indicates expression of the SK2 channel A.

The optical density analysis of SK2 expression using ImageJ software showed that, after 14 days of HH exposure, SK2 expression increased, while GBE significantly reduced its expression B.

Furthermore, to validate the role of GBE in SK2 inhibition, a comparison was made with chemical inhibition of SK2 via apamin. Glutamate and apoptosis markers were studied, and it was found that the GBE-treated group reduces glutamate excitotoxicity Figure 7A and prevents apoptosis Figures 7B,C comparable to a group administered apamin during HH exposure.

This indicates GBE is working through inhibition of SK2 channels during chronic HH exposure for 14 days. FIGURE 7. GBE reduces glutamate excitotoxicity and activates caspase-3 expression via SK2 inhibition. The graphic representation shows that the glutamate level increased on the 14th day of HH exposure, and the glutamate level decreased significantly after GBE treatment.

However, compared with the HH group, the SK2 inhibitor, apamin, can also reduce the toxicity of glutamate A. Immunoblotting showed that compared with the control, the expression of activated caspase-3 increased significantly after 14 days of exposure to HH, while the GBE and SK2 inhibitor, apamin, significantly reduces its expression B.

Densitometry analysis of the activated caspase-3 expression using ImageJ software C. GBE is known to have a multifactorial response. In the present study too, it leads to SK2 inhibition which further inhibits cellular death machinery by modulating downstream signaling as well as activating the survival pathway.

BDNF is well known to promote cell survival and neuroprotection. BDNF reduces the SK2 component by phosphorylation, causes its inactivation, and strengthens synaptic transmission Figure 8A.

We also observed restoration of the depleted BDNF level after GBE treatment during 14 days of HH exposure Figure 8B. Hence, we found it interesting to explore the effect of GBE treatment on ERK activation in the hippocampus during HH exposure.

As shown in Figure 8C , 14 days of HH exposure significantly reduces the ERK activation and, hence, phosphorylation which is further ameliorated by GBE treatment. We have also studied the expression of CaMKII, and it was found that GBE treatment reduces the activity of CaMKII, which indicates that calcium homeostasis is maintained, thus promoting neuroprotection Figure 8D.

Since ERK further activates its downstream transcription factor, i. FIGURE 8. Immunoblot of CREB, p-CREB, ERK, p-ERK, CaMKII, p-CaMKII, and BDNF proteins A. Further densitometry analysis of respective immunoblots was performed through ImageJ software and represented as a bar graph of BDNF B p-ERK C , p-CaMKII D , and p-CREB E.

No significant changes were observed in ERK, CaMKII, and CREB expressions. FIGURE 9. Graphical representation of abstract: Schematic diagram representing the mechanism of action of Ginkgo biloba L.

during HH. The present study was aimed to investigate the efficacy of GBE on HH-induced memory impairment and neurodegeneration in rats. Chronic HH exposure is well documented for memory impairment and neuronal loss in different brain regions especially the hippocampus Maiti et al.

HH conditions have shown detrimental effects on spatial memory Jain et al. The hippocampus is one of the key brain regions involved in spatial memory formation.

Different durations of HH exposure at different altitudes showed significant impairment in spatial memory which got worse in chronic HH exposure conditions Shukitt et al. Similarly, our study also found spatial memory impairment in the MWM test during chronic exposure to HH for 14 days as evident from the increased latency and path length to reach the platform and decreased time spent in the targeted quadrant and number of platform crossing.

On the contrary, GBE that has different beneficial ingredients for mental health showed amelioration in detrimental effect of chronic HH exposure on spatial memory.

Similar observations were reported previously by Vaghef et al. on oxidative stress as well as memory impairments induced by transient cerebral ischemia. GBE has been reported to exhibit anti-inflammatory and neuroprotective properties in other studies Liang et al.

HH causes cellular oxidative damage with consequent damage to lipids, proteins, and DNA that could result in cognitive deficit Maiti et al. Comparable to our study, other studies also substantiate antioxidant efficacy of GBE by reducing oxidative stress and enhancing the antioxidant level in different neurological disorders Lugasi et al.

has different active ingredients that account for its antioxidant property. We further examined neurodegeneration in different groups that shows that GBE expressively reduced the HH-mediated neuronal loss as apparent from the decreased number of Fluoro-Jade B—positive neurons and activated caspase-3—positive neurons, which also augmented after 14 days of HH exposure in the hippocampal tissue.

HH exposure also altered the neuronal morphology as evident from cresyl violet staining; however, administration of GBE resulted in recovery of the neuronal morphology by reducing the number of pyknotic cells in the GBE-treated group; similar findings were observed in another study also Kumari et al.

One of the active ingredients of GBE, i. Calcium overload is known to cause glutamate excitotoxicity and further lead to apoptosis Belov et al. Prevention is reproducible and comparable to the group with apamin-sensitive SK2 inhibition that validates the role of SK2 channels in GBE-mediated neuroprotection.

We further explored the putative signaling pathway involved in GBE-mediated neuroprotection. BDNF is well known to promote cell survival and neuroprotection in control conditions as well as in different pathological conditions Sheng et al. A study by Kramár et al.

showed that BDNF decreases the SK2 component by phosphorylation, causes its inactivation, and strengthens synaptic transmission Kramar et al. Therefore, we postulated that GBE-mediated SK2 inhibition may work through the BDNF-mediated signaling mechanism, and it was observed that GBE treatment increases the BDNF level during HH exposure in comparison with groups exposed to HH alone.

Hence, in the present study, we demonstrated that BDNF further inhibits CaMKII phosphorylation and activates CREB through the ERK pathway.

GBE prevents chronic HH-induced memory impairment and oxidative stress. It further averts neurodegeneration by reducing apoptosis. The neuroprotective effect of GBE may contribute to its efficacy in facilitating BDNF overexpression that further inhibits SK2 channels and reduces cell death.

NiK conceived and coordinated the study. NiK, VJ, and NeK performed and analyzed the experiments. NiK and NeK wrote the manuscript. MK, AD, and RK contributed with hypobaric hypoxia exposures and sample collection. DP and BK supervised the study and reviewed the manuscript before submission.

All authors have read and approved the final version of the manuscript. The study was supported by a grant No. DIP NK from DRDO, Ministry of Defense, Government of India. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The authors are thankful to Usha Panjwani for generous support during various stages of this work and R. Tirpude for support with experimental animals.

leaf extract; GSH, glutathione; HA, high altitude; HH, hypobaric hypoxia; MDA, malondialdehyde; PFA, paraformaldehyde; SK channels, small conductance calcium-activated potassium channels; ROS, reactive oxygen species.

Abdel-Wahab, B. Ginkgo Biloba Protects against Intermittent Hypoxia-Induced Memory Deficits and Hippocampal DNA Damage in Rats. Phytomedicine 19 5 , — PubMed Abstract CrossRef Full Text Google Scholar. Achete de Souza, G. Effects of Ginkgo Biloba on Diseases Related to Oxidative Stress. Planta Med.

Adelman, J. Agell, N. Cell Signal. Anjum, V. Antithrombocytopenic and Immunomodulatory Potential of Metabolically Characterized Aqueous Extract of Carica Papaya Leaves. Aydin, D. Effects of Ginkgo Biloba Extract on Brain Oxidative Condition after Cisplatin Exposure.

Basnyat, B. High-altitude Illness. The Lancet , — CrossRef Full Text Google Scholar. BelovKirdajova, D. Ischemia-Triggered Glutamate Excitotoxicity from the Perspective of Glial Cells. Front Cel Neurosci 14, Belviranl, M. The Effects of Ginkgo Biloba Extract on Cognitive Functions in Aged Female Rats: the Role of Oxidative Stress and Brain-Derived Neurotrophic Factor.

Brain Res. Blecharz-Klin, K. Pharmacological and Biochemical Effects of Ginkgo Biloba Extract on Learning, Memory Consolidation and Motor Activity in Old Rats. Acta Neurobiol. Wars 69, — PubMed Abstract Google Scholar.

Chandrasekaran, K. Bilobalide, a Component of the Ginkgo Biloba Extract EGb , Protects against Neuronal Death in Global Brain Ischemia and in Glutamate-Induced Excitotoxicity. Cel Mol Biol Noisy-le-grand 48, — Google Scholar.

Chen, J. Intermittent Hypoxia Protects Cerebral Mitochondrial Function from Calcium Overload. Acta Neurol. Dröge, W. Free Radicals in the Physiological Control of Cell Function. Gaidin, S. The Selective BDNF Overexpression in Neurons Protects Neuroglial Networks against OGD and Glutamate-Induced Excitotoxicity.

Goor, F. Autocrine Regulation of Calcium Influx and Gonadotropin-Releasing Hormone Secretion in Hypothalamic Neurons. Cel Biol.