RU2245673C2 - Method for detecting both functional and metabolic state of nervous tissue - Google Patents

Abstract

FIELD: medicine, neurology, psychopathology, neurosurgery, neurophysiology, experimental neurobiology.

SUBSTANCE: one should simultaneously register electroencephalogram (EEG) to detect the level of constant potential (LCP). At LCP negativization and increased EEG power one should detect depolarizational activation of neurons and enhanced metabolism. At LCP negativization and decreased EEG power - depolarized inhibition of neurons and metabolism suppression. At LCP positivation and increased EEG power - either repolarized or hyperpolarized activation of neurons and enhanced metabolism. At LCP positivation and decreased EEG power - hyperpolarized suppression of neurons and decreased metabolism of nervous tissue. The method enables to correctly detect therapeutic tactics due to simultaneous LCP and EEG registration that enables to differentiate transition from one functional and metabolic state into another.

EFFECT: higher accuracy of diagnostics.

5 dwg, 1 ex, 1 tbl

Description

The alleged invention relates to medicine, namely to neurology, psychopathology, neurosurgery, neurophysiology and experimental neurobiology, and is intended to determine the functional and metabolic state of the nervous tissue.

A known method for determining the state of nerve tissue by registering cerebral blood flow by the method of hydrogen clearance using platinum electrodes implanted in brain tissue or venous sinus / 1 /. However, the known method only indirectly allows you to judge the change in the functional and metabolic state of the brain and does not allow to differentiate many pathological and physiological conditions. The method is uninformative, difficult to implement, highly invasive and of little use in the clinic.

A known method for determining the functional and metabolic state of nerve tissue by conducting positron emission tomography of the brain / 2 /. The disadvantage of this method is the difficulty of implementation, high economic costs and the use of expensive equipment for the study, the need for preliminary preparation of patients for the study. The disadvantages of the method are the impossibility of dynamic monitoring of the state of the brain during medical manipulations in the clinic, the complexity and inconvenience for implementation in experimental studies on small laboratory animals.

A known method of recording the level of constant potential (SCP) / 3 / and electroencephalogram / 4 /. Shifts of SCP reflect polarization processes in the nervous tissue / 5 /. Positive AMP shifts accompany the development of polarization processes (repolarization or hyperpolarization of neurons and glial cells, a decrease in the extracellular concentration of K (+) ions and an increase in the concentration of Ca (2+) and Na (+) ions. Negative AMP shifts reflect the development of depolarization processes (depolarization of neurons and glial cells, an increase in the extracellular concentration of K +, a decrease in the concentration of Ca (2+) and Na (+)). However, the determination of polarization processes does not allow us to differentiate and record the transition from one functional metabolic state of the nervous tissue in the other. The method enables to determine not pathological and physiological processes, has a low accuracy of diagnosis and little information, which limits its application in clinical and experimental.

Closest to the proposed method is a method of recording the total slow electrical activity of the brain from the surface of the head (EEG) / 6 /. It is known that activation processes in the nervous system are accompanied by depression of alpha activity. The development of pathological conditions in connection with metabolic disturbances, as in brain ischemia, is associated with the appearance of slow-wave activity of the theta and delta ranges. Inhibition of the functional state with deepening hypoxia and ischemia leads to depression of the EEG / 7 /, / 8 /, / 9 /. Despite the fact that this method has a number of positive properties, it does not allow subtly differentiating many physiological and pathological FS.

Thus, at present, there are no separate methods for assessing the functional and metabolic state of nervous tissue in the entire range of physiological phenomena.

The objective of the proposed invention is to provide a method that improves the accuracy of determining the functional and metabolic state of the nervous tissue, by differentiating and recording the transition of one functional and metabolic state to another.

The problem is achieved in that in the known method for determining the functional and metabolic state of nerve tissue by recording its bioelectric activity, at the same time as the total slow electrical activity (EEG), the level of constant potential (PP) is recorded, and they are compared by the nature of the change in the measured parameters in one and the same period of time determine the functional and metabolic state of the nervous tissue.

What is new in achieving the set technical result is that in parallel with recording the level of constant potential, the total slow electrical activity of the nervous tissue is recorded and the functional and metabolic state of the nervous tissue is determined by changing the measured parameters. Simultaneous registration of the level of constant potential and total slow electrical activity allows the assessment of polarization and activation processes in the nervous tissue, which increases the accuracy of determining the functional and metabolic state due to differentiation and registration of the transition of one functional and metabolic state of the nervous tissue to another.

Negatiation of SCP and an increase in the power of EEG rhythms are considered as the development of depolarization activation of neurons and increased metabolism of nerve tissue.

Negatization of PP and a decrease in the power of EEG rhythms are considered as the development of depolarization inhibition and inhibition of the metabolism of nerve tissue.

Positivization of PP and an increase in the power of EEG rhythms are considered as the development of repolarization or hyperpolarization activation of neurons and increased metabolism of nerve tissue.

Positivization of PP and a decrease in the power of EEG rhythms are considered as the development of hyperpolarizing inhibition of neurons and weakening of the metabolism of nervous tissue.

The method is as follows.

At the object of study, non-polarizing (silver chloride) electrodes using a DC amplifier conduct simultaneous registration of the level of constant potential and the total slow electrical activity in the studied nervous tissue.

The functional and metabolic state of the brain is determined by enhancing or inhibiting the total slow electrical activity and positive or negative shifts in the level of constant potential.

With positive shifts in SCP and inhibition of EEG power, the development of cell membrane polarization (re- or hyperpolarization) and a decrease in the metabolic need of nerve tissue are determined. This functional state develops either with hyperpolarization inhibition, or with the return of the membrane potential to the level of resting potential after depolarization exaltation of excitability.

With positive shifts in SCP and an increase in EEG power, the development of cell membrane polarization (re- or hyperpolarization) and the optimal level of nervous tissue metabolism are determined. This functional state develops either with hyperpolarization exaltation of excitability, or with repolarization in connection with the release of cells from cathodic depression or parabiosis.

With negative changes in SCP and an increase in EEG power, the development of depolarization is determined, accompanied by an increase in the excitability and metabolic demand of the nervous tissue. These changes are observed, in particular, during the development of the activation reaction.

With negative changes in SCP and inhibition of EEG power, the development of depolarization inhibition (by parabiotic or cathodic type) and a decrease in metabolism are determined.

Example.

To assess the functional and metabolic state of the nervous tissue, our method was used to study AMR and EEG in modeling acute circulatory ischemia of different depths. Ischemia was simulated in two ways on the same rats sequentially during one experiment. Preliminarily, 2-3 days before the experiment, outbred white rats weighing 150-200 g of both sexes (n = 12) under nembutal anesthesia under the bones of the skull over the frontal cortex of the right and silver chloride electrodes were implanted in the left hemisphere. An indifferent silver chlorinated electrode was placed in the bones above the frontal sinuses. Electrode leads were attached to the skull with quick-hardening plastic.

After that, the rats were operated under nembutal anesthesia (40 mg / kg) for modeling ischemia. The registration of brain bioelectric activity according to a unipolar technique was started before anesthesia was introduced and continued throughout the experiment using a multichannel DC amplifier with an input impedance of 1 MΩ and a frequency bandwidth of 0-40 Hz. The data were digitized (100 Hz) and entered into a computer for further processing. The determination of the spectrum of rhythms and its power was carried out using the Fourier transform. In this case, five ranges were distinguished: delta-1- (0.2-1 Hz), delta-2- (1-4 Hz), theta (4-8 Hz), alpha (8-13 Hz) and beta (13-30 Hz) rhythm. SCP was averaged over periods corresponding to epochs of EEG analysis.

The first model of ischemia (“Ischemia-1”) consisted in bandaging both common carotid arteries. The duration of the isolated action of this model of ischemia was 20 minutes, after which an additional introduction of rat occluder into the middle cerebral artery (MCA) of the left hemisphere (Ischemia-2) / 10 / was carried out. Combined modeling of two types of circulatory ischemia was carried out for 60 minutes, after which the occluder was removed and SCP and EEG were recorded for another 16 minutes.

In a separate series of experiments (n = 10), an investigation was made of AMR and EEG power after intracerebroventricular administration of cyclopentyladenosine (CPA). CPA was administered under anesthesia (Nembutal 40 mg / kg) by injection of the drug (25 μg / kg) into the middle cerebral ventricles of the right hemisphere.

Thus, it was possible to assess the nature of changes in the complex of bioelectric indicators (EEG and SCP) during anesthesia with ethanol-sodium, intracerebroventricular administration of CPA, ligation of the common carotid arteries, MCA occlusion and reperfusion and, based on literature data and the patterns of changes in biopotentials obtained by us, determine functional and metabolic state of the brain.

The results were processed statistically using the methods of parametric and nonparametric statistics for dependent and independent samples: Student's t test, sign test, Wilcoxon test, Wilcoxon-Mann-Whitney test.

Figure 1 shows the change in the power of the EEG and SCP during the modeling of ischemia in two ways. It is seen that the ligation of the common carotid arteries (“Ischemia 1”) led to an increase in the EEG power in the frontal and parietal cortex of the right and left hemispheres. The increase in the power of the rhythm spectrum in all leads as a whole for the entire rat sample was 14.04 ± 1.75% (p <0.001). The largest increase in amplitude (20.04 ± 3.2%) was observed in the alpha range, where it increased from 23.71 ± 0.86 to 28.46 ± 1.09 μV (p <0.001). For the theta rhythm, the increase was 16.31 ± 3.15% and it changed from 46.52 ± 2.03 to 54.01 ± 2.08 μV (p <0.01). The increase in the amplitude of the delta rhythm was 10.61 ± 2.79%: from 106.85 ± 3.96 to 118.38 ± 3.74 μV (p <0.01). The beta rhythm has changed the least. Its power increased only by 9.21 ± 4.33% from 7.31 ± 0.34 to 7.98 ± 0.33 μV. Nevertheless, this increase was statistically significant (paired Student's t-test showed differences at p <0.01). Simultaneously with an increase in the power of EEG rhythms, a slight negative deviation of the SCP was observed, which by the end of the period was 1222.51 ± 290.1 μV (p <0.01). According to the available literature, the state of neural activation is accompanied by an increase in the metabolism of nervous tissue / 12 /. A negative shift in PP indicates the depolarization of neural elements / 9 /, / 7 /, and, in general, with an increase in neural activity, this indicates the development of a functional state like a cathelectron in nerve cells.

The additional introduction of the occluder in the left hemisphere SMA against the background of the development of general inhibition of EEG rhythms led to more significant changes in the SCP in the neocortex, which had both a positive and a negative orientation (Fig. 1, “Ischemia-2”). In the left hemisphere, both in the frontal and parietal cortex, a significant (several tens of millivolts) negative deviation of the SCP was observed. The average inhibition of EEG rhythms in the left hemisphere as a whole in the sample amounted to 25.62 ± 2.41 in the frontal cortex and 22.74 ± 1.94% in the parietal cortex. Most of all, the changes affected the slow frequencies (see table). In the right hemisphere, the inhibition of rhythms was significantly less than in the left hemisphere, and amounted to 14.28 ± 2.49% for the frontal cortex and 13.03 ± 2.19% for the parietal cortex. Analysis of EEG changes in the right hemisphere separately for frequencies (see table) shows that inhibition affected only the delta rhythm. In the remaining frequency ranges, the introduction of the occluder into the left hemisphere SMA did not lead to significant changes in rhythms compared with the period preceding brain ischemia. Differences in changes in biopotentials in the right hemisphere also affected SCP. In the parietal cortex of the right hemisphere, the negative deviation was almost 3 times less than in the left hemisphere (see figure 2). In the frontal cortex of the right hemisphere, the deviation of the constant potential was generally positive.

Thus, the development of circulatory ischemia according to model 1 was accompanied by a relatively small negative deviation of SCP and an increase in the power of EEG rhythms in all leads. The use of left hemisphere SMA in model 2 of occlusion led to significant additional negativity of the left hemisphere AMR and inhibition of EEG in it against this background. According to the literature / 12, 13, 14, 15 /, negative SCP deviation and EEG depression are indicators of the development of deep brain ischemia. This allows us to consider that in the left hemisphere during Ischemia-2, severe cerebral ischemia was modeled, which is also confirmed by histological analysis / 10 /. The changes in the EEG and SCP observed in the right hemisphere in Ischemia-2 are most likely associated with the redistribution of blood flow due to the activation of collateral circulation mechanisms. In the literature there is evidence of an increase in stroke blood flow in the pool of symmetrical arteries of the opposite hemisphere stroke / 16 /. The positive changes in SCP observed in the frontal cortex of the right hemisphere indicate an increase in blood flow in this part of the brain, and a decrease in the power of EEG rhythms reflects an improvement in the metabolic and functional state of the nervous tissue, worsened during the period of Ischemia-1. In other words, a condition close to preoperative was formed in the frontal cortex of the left hemisphere. Due to increased blood supply to the frontal cortex, most likely, there was a partial "robbery" of the parietal cortex of the same hemisphere. Therefore, in the parietal cortex of the right hemisphere, after the introduction of the occluder into the MCA of the left hemisphere, the SCP was also negative. A negative shift in the SCP in the parietal cortex of the right hemisphere, coupled with a slight decrease in the rhythm power in it, indicates that a relatively unfavorable functional state of the nervous tissue was also formed here during Ischemia-2.

A comparison of the results of changes in the biopotentials of the two models of ischemia allows us to consider Ischemia-1 as a model of weaker ischemia. A similar model of ischemia showed / 17 / that, despite the inactivation of the barrier-transport systems of the blood-brain barrier, neurons maintain a high level of oxidative metabolism due to the existence, in particular, of intracellular compensatory-adaptive mechanisms. According to published data, on the hypoxic effect of neurons correspond to the primary reaction of depolarization of the resting potential and activation of impulse activity, which is replaced by a parabiotic type as hypoxia deepens y / 18 /. Apparently, the activation of EEG and the negativity of SCP, observed during ligation of the common carotid arteries, and reflect the primary exaltation of the excitability of brain cells according to the catelectronic type.

Extraction of the occluder and reperfusion of the brain by SMA caused an increase in the power of EEG rhythms and a positive deviation of SCP in the frontal and parietal cortex of the left hemisphere. A similar pattern was observed in the parietal cortex of the right hemisphere. In the frontal cortex of the right hemisphere, reperfusion of the SMA of the left hemisphere led to an increase in EEG power against the background of negative SCP. In other words, the restoration of the blood flow regime corresponding to the period of “Ischemia-1” in the left hemisphere, according to electrophysiological data, reflects an improvement in the metabolic and functional state of the nervous tissue. Similar processes took place in the parietal cortex of the right hemisphere, while, in the frontal cortex, extraction of the occluder, reducing collateral blood supply, worsened its metabolic state, which activated it again.

Reperfusion data indicate that brain activation observed by EEG is possible against a background of both negative and positive abnormalities of SCP. If in the first case this indicates a deterioration of the functional and metabolic state (as well as with Ischemia-1), then in the second case it indicates an improvement in connection with the exit from the state of cathodic depression with improved blood flow.

Thus, the use of two models of ischemia showed that the development of ischemic processes associated with a deterioration in the functional and metabolic state of the neocortex is accompanied by ambiguous changes in the EEG. With relatively weak ischemia, the negativeness of SCP, which, as is known, reflects an increase in the depolarization of the neuroglial complex / 7 /, / 9 /, is accompanied by activation of the EEG. With the deepening of ischemia, there is an even greater negativity of SCP and depression of EEG rhythms, which reflects the development of parabiotic inhibition in the nervous tissue. A positive deviation of AMR and an increase in EEG power is observed in the opposite process: with repolarization of neurons and their exit, most likely, from the state of parabiosis in connection with the improvement of blood supply to the nervous tissue and its metabolic state.

Figure 3 shows the change in SCP and EEG in rats after intracerebroventricular administration of CPA in 4 leads. It is seen that the injection of CPA caused in all cases a positive deviation of AMR with primary activation of the power of most (delta-1, delta-2 and beta) EEG rhythms, which was then replaced by their depression. The greatest positivization of SCP was observed in the right hemisphere, i.e. on the side of the drug. According to published data / 19 /, / 20 /, adenosine and its analogues (which include CPA) inhibit the impulse activity of neurons and cause membrane hyperpolarization. The positivization of SCP obtained in our experiment also indicates that this deviation reflects the hyperpolarization of neurons. Moreover, hyperpolarization can be combined both with an increase in the power of rhythms and their inhibition. The primary activation of the EEG (Fig. 4) in the delta and beta ranges with a positive shift in SCP indicates, most likely, the development of a functional state similar to anodic exaltation, with an increase in metabolic demand against the background of the maintenance of ionic homeostasis, which changes rather quickly to hyperpolarization inhibition . The developed inhibition of rhythm power after the introduction of CPA was stable and took place during 60 minutes of observation.

Evaluation of the functional and metabolic state of the nervous tissue by our method was also carried out after administration of ethaminal sodium. The experimental design was as follows: after connecting the connectors with wires from the amplifier to the terminals of the electrodes mounted on the head of the rat, the animal was placed in an experimental chamber, where after calming it down for 5-10 minutes the initial recording of biopotentials was carried out, then the rat was taken into hands and using Ethamine-sodium (40 mg / kg) was intraperitoneally injected into the syringe. Figure 5 shows the change in the recorded biopotentials with intraperitoneal administration to ethinal-sodium rats. It is seen that immediately after the injection of the drug, a negative deviation of the SCP was observed (p <0.001) and an increase in the power of the EEG (p <0.001). After 1-2 minutes, a positive shift in constant potential appeared while maintaining increased rhythm power (Fig. 5. “Preson”). The loss of pain sensitivity and falling asleep of the animal occurred against the background of further positivization of SCP, while reducing the amplitude of the EEG rhythms, the inhibition of which reached a maximum with deepening sleep.

Integrated registration of AMR and EEG during anesthesia, thus, allowed to identify at least three consecutive stages of changes in the functional and metabolic state of the brain. Negative emotional arousal, which develops evidently in animals during needle piercing of the skin and the administration of the drug, was accompanied by the negation of the constant potential and an increase in EEG power, reflecting, apparently, the development of depolarization exaltation of neuronal excitability and increased metabolism of neural tissue. As ethinal sodium was absorbed into the blood, a positive deviation of the SCP appeared while maintaining increased EEG power. The nature of the change in constant potential indicates the development of re- and hyperpolarization processes. The increased amplitude of the rhythms indicates the absence of hyperpolarizing inhibition even at that time. In the literature there is evidence of hyperpolarization changes in the membrane potential of neurons under the action of Nembutal / 21 /. A comparison of SCP and EEG characteristics allows us to consider the development of neurons in the “Preson” period of a functional state corresponding to anodic exaltation and an increase in nervous tissue metabolism during this period. Finally, the onset and development of narcotic sleep was accompanied by an even stronger positive shift in constant potential and inhibition of EEG power. This functional state reflects, apparently, the deepening of cell hyperpolarization and the onset of hyperpolarizing inhibition with a decrease in the metabolism of nerve tissue.

Comparison of these data with the results of changes in biopotentials with the introduction of CPA indicates their fundamental similarity. In either case, a positive deviation of SCP was accompanied by initial activation of EEG with its subsequent inhibition. This indicates that in both cases, generally similar functional and metabolic processes developed.

The results of the presented experiments demonstrated high diagnostic capabilities of the proposed method. Separately, neither EEG nor AMP allow a fine differentiation of the functional and metabolic state of the nervous tissue. The advantage is also the relative simplicity of the technique. The proposed method allows to detect and differentiate the transition of one functional and metabolic state to another, thereby increasing the accuracy of diagnosis of pathological and physiological conditions, allows for adequate drug therapy of pathological conditions, for example, with ischemia, aimed at restoring the functional and metabolic state of the nervous tissue, and determine prognosis and correctness of treatment after a particular therapy, as well as to study the effect of extreme factors on or anizm person.

Thus, the proposed method makes it possible to accurately determine the functional and metabolic state of the nervous tissue, adequately differentiate physiological and pathological conditions, to register the transition from one functional and metabolic state to another, which increases the accuracy of diagnosis and high information content of the method and allows you to correctly determine the tactics of treatment in neurology , psychopathology, neurosurgery and neurophysiology, and also allows the development of new pathogenetic neuroses rotor drugs and study the mechanisms of pathological and physiological conditions in the experiment.

Sources of information taken into account

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Table
The change in the power of rhythms of various ranges in% during the period of "Ischemia-2" in relation to the period preceding "Ischemia-1" Left hemisphere Right hemisphere Delta 1 Delta 2 Theta Alpha Beta Delta 1 Delta 2 Theta Alpha Beta Frontal bark -50.83 ± 3.52 -38.10 ± 5.07 -34.10 ± 2.41 -11.76 ± 4.09 -8.45 ± 6.22 -51.86 ± 2.44 -23.45 ± 3.17 + 3.45 ± 2.20 + 1.39 ± 2.62 -7.18 ± 5.94 Parietal cortex -44.84 ± 4.38 -21.77 ± 3.31 -21.17 ± 3.31 -13.03 ± 2.90 -17.09 ± 5.20 -42.94 ± 1.60 -18.35 ± 3.24 -4.94 ± 3.39 -2.63 ± 3.43 -4.23 ± 1.89 "-" decrease, "+" - increase in the amplitude of rhythms

Claims (1)

A method for determining the functional and metabolic state of nerve tissue, including registration of an electroencephalogram (EEG), characterized in that at the same time as the EEG, the level of constant potential (SCP) is recorded, and with the negativeness of the SCP and increasing the power of the EEG, the depolarization activity of neurons and increased metabolism are determined; with negative SCP and a decrease in EEG power, depolarization inhibition of neurons and inhibition of metabolism; with positivization of AMR and an increase in EEG power, repolarization or hyperpolarization activation of neurons and increased metabolism; with positivities of SCP and a decrease in EEG power, hyperpolarization inhibition of neurons and a decrease in the metabolism of nerve tissue.

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