Application of hyperbaric oxygen-induced sodium channel active marker in epilepsy susceptibility evaluation

Abstract

The invention discloses an application of a hyperbaric oxygen-induced sodium channel active marker in epilepsy susceptibility evaluation. Belongs to the technical field of biomedicine. In the application, a sodium channel active marker comprises peripheral blood sodium channel protein, peripheral blood phosphorylated sodium channel protein, neuron-specific enolase, S100B protein and epilepsy discharge frequency. The high-pressure oxygen induction condition is that the pressure is increased to 2.0 absolute atmospheric pressure within 15 min, then pure oxygen is absorbed for 1 h under the stable pressure condition, and finally the pressure is reduced to normal pressure at a constant speed within 15 min. The method has the advantages of being high in evaluation accuracy and small in harm to human bodies, and the problem that in the prior art, epilepsy susceptible high-risk groups cannot be screened is effectively solved.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of a hyperbaric oxygen-induced sodium channel activity marker in the evaluation of epilepsy susceptibility. Background Technology

[0002] Epilepsy is a common chronic neurological disorder characterized by sudden abnormal electrical discharges in the brain. Its core pathophysiological mechanisms are closely related to increased neuronal excitability and abnormal sodium ion channel function. Currently, the diagnosis of epilepsy primarily relies on clinical symptoms and electroencephalogram (EEG) recordings during seizures. However, for assessing susceptibility to epilepsy—that is, predicting the future risk of developing epilepsy in an individual who has not yet experienced seizures—there is a lack of effective and objective biological indicators in clinical practice. Summary of the Invention

[0003] To address the aforementioned shortcomings in existing technologies, this invention provides an application of hyperbaric oxygen-induced sodium channel activity markers in the evaluation of epilepsy susceptibility. This method has the advantages of high evaluation accuracy and minimal harm to the human body, effectively solving the problem that existing technologies cannot screen high-risk groups susceptible to epilepsy.

[0004] To achieve the above objectives, the technical solution adopted by the present invention to solve its technical problem is as follows: Application of a hyperbaric oxygen-induced sodium channel activity marker in the evaluation of epilepsy susceptibility.

[0005] Furthermore, the sodium channel activity markers include peripheral blood sodium channel proteins, peripheral blood phosphorylated sodium channel proteins, neuron-specific enolases, and S100B proteins.

[0006] Furthermore, the specific testing method is as follows: Furthermore, the high-pressure oxygen induction conditions are as follows: pressurize to 2.0 atmospheres absolute within 15 minutes, then inhale pure oxygen for 1 hour under stable pressure, and finally depressurize to atmospheric pressure at a constant rate within 15 minutes.

[0007] Furthermore, the evaluation criteria are as follows: if the change value of the biomarker is greater than the threshold, the individual is judged to be at high risk of epilepsy.

[0008] Furthermore, the specific testing method is as follows: a. Baseline detection: Before an individual receives hyperbaric oxygen exposure, biological samples are collected to detect the levels of one or more biomarkers related to neuronal sodium channel activity, denoted as baseline value B0; b. Hyperbaric oxygen exposure: The individual receives a standard treatment regimen of hyperbaric oxygen exposure, the parameters of which are: slowly pressurize for 15 minutes to a pressure of 2.0 atmospheres absolute, inhale pure oxygen at a constant pressure for 60 minutes, and then gradually reduce the pressure to normal within 15 minutes. c. Post-exposure detection: Within a specific time window after the end of hyperbaric oxygen exposure (set as 2 hours after exposure), the level of the biomarker was detected again and recorded as the post-exposure value P1; d. Susceptibility calculation: Calculate the change value of the biomarker Δ = P1 - B0 or ​​the change rate (P1 - B0) / B0 × 100%), compare the change value with a preset threshold, and if the change value exceeds the threshold, the individual is determined to be a high-risk individual for epilepsy.

[0009] The beneficial effects of this invention are as follows: Hyperbaric oxygen therapy (HBOT) is a method of treating diseases by inhaling high concentrations of oxygen at an environment above one atmosphere. It is widely used for conditions such as cerebral ischemia, carbon monoxide poisoning, and sudden deafness, and is believed to exert a protective effect by increasing blood oxygen content and oxygen partial pressure. However, based on an animal experimental project, the inventors discovered using patch-clamp techniques that hyperbaric oxygen can increase the activity of neuronal voltage-gated sodium channels, which may be a key molecular mechanism leading to increased neuronal excitability and ultimately inducing epilepsy. Further investigation revealed that hyperbaric oxygen exposure with specific parameters does not always have a protective effect; instead, it can act as a controllable physiological stressor, specifically revealing a functional "hypersensitive" state of neuronal sodium channels, a state closely related to susceptibility to epilepsy. This finding contradicts the traditional understanding that hyperbaric oxygen is a neuroprotective agent. Therefore, the idea arose: using a standardized hyperbaric oxygen exposure commonly used in clinical treatment as a diagnostic stimulus could safely and effectively "excite" latent neuronal hyperexcitability. By quantifying the dynamic changes of specific biomarkers before and after stimulation, an active and quantitative assessment of epilepsy susceptibility was achieved, solving the problem that existing technologies can only perform passive diagnosis after an epileptic seizure. Attached Figure Description

[0010] Figure 1 A statistical graph showing the maximum current density of neurons 2 in the CA1 region of the hippocampus of rats with cerebral ischemia at different time points in three hyperbaric oxygen therapy groups. Figure 2 The IV curves for each group of sodium currents are shown. Figure 3 The graph shows the steady-state activation curves of sodium channels in each group. Detailed Implementation

[0011] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention; that is, the described embodiments are merely some embodiments of the invention, and not all embodiments.

[0012] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0013] It should be noted that relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes the element.

[0014] The features and performance of the present invention will be further described in detail below with reference to experimental examples and accompanying drawings.

[0015] This invention is based on key findings obtained by combining the following experimental examples and figures: Test case 1. Animal model establishment: A rat model of cerebral ischemia was established using the classic middle cerebral artery occlusion (MCAO) method.

[0016] 2. Grouping: Rats were randomly divided into a cerebral ischemia model group (MCAO 6H, MCAO 24H, MCAO 48H, MCAO 72H) and a hyperbaric oxygen therapy group (HBOT 1 session, HBOT 6 sessions, HBOT 12 sessions). The hyperbaric oxygen therapy group received hyperbaric oxygen therapy with specific parameters (2.0 ATA, 60 minutes / session, once daily) after model establishment.

[0017] 3. Core Detection Methods: After treatment, neurons in the CA1 region of the hippocampus of rats in each group were acutely isolated. Using whole-cell patch-clamp technique, voltage-gated sodium currents in the CA1 region of the hippocampus of each group were recorded. The activation threshold and peak current of the sodium current were observed, the current density was calculated, and current-voltage (IV) curves and steady-state activation curves were plotted.

[0018] 4. Results showed that after one HBOT treatment, compared with the MCAO 6H group [(-36.84±3.53) pA / pF], the MCD of the MCAO 48H group [(-25.49±3.29) pA / pF] and the MCAO 72H group [(-21.13±1.23) pA / pF] were significantly reduced, with statistically significant differences (P<0.01, P<0.001). After 12 HBOT treatments, compared with the MCAO 6H group [(-47.18±9.86) pA / pF], the MCD of the MCAO 72H group [(-13.96±5.57) pA / pF] was significantly reduced, with statistically significant differences (P<0.05). The sodium current MCD of hippocampal neurons in all four MCAO rat groups showed a decreasing trend.

[0019] 5. Conclusion: With prolonged ischemia-reperfusion time, some sodium channels become inactivated or close, resulting in a decrease in sodium current. After a prolonged period of ischemia-reperfusion injury, some central neurons suffer irreversible damage or even death, leading to a reduction in the number of cell membrane sodium channels or even their inability to generate normal sodium current, thus causing a decrease in sodium current (see...). Figure 1 In the four groups of cerebral ischemia rats, compared with 1 HBOT, the IV curve shifted to the right after 6 HBOTs, indicating that the cell membrane permeability to sodium was reduced, requiring a higher voltage to achieve the same current; after 12 HBOTs, the IV curve shifted to the left, which was more obvious in the MCAO 6H group, indicating that the cell membrane permeability to sodium was increased. It is speculated that the rats in the MCAO 6H group may be in the reperfusion injury stage. However, with the increase of HBOTs, the cell membrane permeability to sodium increased, and the activity of sodium channels gradually recovered. Moreover, the earlier the intervention time, the better the protective effect on sodium channels in central nerve cells (see...). Figure 2 After 12 HBOT treatments, the sodium current activation curves of rats in the MCAO 6H and MCAO 24H groups shifted towards hyperpolarization, with the shift being more pronounced in the MCAO 6H group (see...). Figure 3 This indicates that hyperbaric oxygen therapy increases sodium current in neurons of the CA1 region of the hippocampus in rats with cerebral ischemia (manifested as an increase in peak sodium current density, a leftward shift of the IV curve, and a shift of the steady-state activation curve toward hyperpolarization), which enhances the activity of sodium ion channels, making them easier to activate and putting them in a "hypersensitive" state.

[0020] Patch-clamp experiments directly confirmed that hyperbaric oxygen can enhance neuronal sodium channel activity. Changes in the levels of sodium channel proteins (possibly derived from neuronal exosomes), neuron-specific enolase (NSE), and S100B protein (a marker of astrocyte activation / damage) in peripheral blood are all direct / indirect, quantifiable reflections of this neuronal hyperexcitability at the systemic level. Therefore, detecting these markers can be used as an alternative to invasive patch-clamp techniques to achieve non-invasive / minimally invasive assessment of clinical susceptibility.

Claims

1. Application of a hyperbaric oxygen-induced sodium channel activity marker in the evaluation of epilepsy susceptibility.

2. The application as described in claim 1, characterized in that, The sodium channel activity markers include peripheral blood sodium channel proteins, peripheral blood phosphorylated sodium channel proteins, neuron-specific enolases, and S100B proteins.

3. The application as described in claim 1, characterized in that, The high-pressure oxygen induction conditions are as follows: pressurize to 2.0 atmospheres absolute within 15 minutes, then inhale pure oxygen for 1 hour under stable pressure, and finally depressurize to normal pressure at a constant rate within 15 minutes.

4. The application as described in claim 1, characterized in that, The evaluation criteria are as follows: if the change value of the biomarker is greater than the threshold, the individual is judged to be at high risk of epilepsy.

5. The application as described in claim 1, characterized in that, The specific testing method is as follows: a. Baseline detection: Before an individual receives hyperbaric oxygen exposure, biological samples and / or physiological signals are collected to detect the levels of one or more biomarkers related to neuronal sodium channel activity, denoted as baseline value B0; b. Hyperbaric oxygen exposure: The individual receives a standard treatment regimen of hyperbaric oxygen exposure, the parameters of which are: slowly pressurize for 15 minutes to a pressure of 2.0 atmospheres absolute, inhale pure oxygen at a constant pressure for 60 minutes, and then gradually reduce the pressure to normal within 15 minutes. c. Post-exposure detection: Within a specific time window after the end of hyperbaric oxygen exposure, the levels of the biomarkers are detected again and recorded as the post-exposure value P1; d. Susceptibility calculation: Calculate the change value of the biomarker Δ = P1 - B0 or ​​the change rate (P1 - B0) / B0 × 100%), compare the change value with a preset threshold, and if the change value exceeds the threshold, the individual is determined to be a high-risk individual for epilepsy.

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