ISSN 0006-3509, Biophysics, 2021, Vol. 66, No. 6, pp. 956–964. © Pleiades Publishing, Inc., 2021.

Russian Text © The Author(s), 2021, published in Biofizika, 2021, Vol. 66, No. 6, pp. 1123–1132.

CELL BIOPHYSICS

Hypothesis on Pollution of Neuronal Membranes,

Epilepsy and Ketogenic Diet
Yu. D. Nechipurenkoa, *, R. C. Garcia Reyesb, and J. L. Hernandez Caceresb, **
a Institute of Molecular Biology “V.A. Engelhardt”, Russian Academy of Sciences, Moscow, 119991 Russia
b
Cuban Center for Neurosciences, Playa, Havana, 15202 Cuba
*e-mail: nech99@mail.ru
**e-mail: jose.caceres@cneuro.edu.cu
Received July 1, 2021; revised August 30, 2021; accepted August 31, 2021

Abstract—Taking into account recent facts, Altrup’s neuron’s membrane pollution hypothesis for epilepsy is
dealt with. This hypothesis links paroxysmal depolarization shifts observed during epileptic activity, and sin-
gle-neuron pacemaker potentials. Membrane’s physicochemical characteristics, fluidity and pollution influ-
ence on its capability to conduct impulses and polarize. Previously used means of epilepsy treatment based
on the ketogenic diet, as well as their possible mechanisms are discussed on the light of Altrup’s hypothesis.
Among possible action mechanisms for ketogenic diet, we underline ketone bodies antiepileptic action, the
role of increased synthesis of glutathione and the effect of polyunsaturated fatty acids (PUFA) and cholesterol
as components included into the ketogenic diet. These three mechanisms, among others, lead to a regulation
of fluidity and other biophysical properties of the membrane bilayer as well as to a cleansing of the membrane
from amphiphilic polluters, in accordance with Altrup’s hypothesis.

Keywords: epileptic activity, paroxysmal depolarization shifts, pacemaker potential, Altrup’s hypothesis,
membrane pollution, lateral pressure, ketogenic diet, glutathione, polyunsaturated fatty acids
DOI: 10.1134/S0006350921060129

Nearly 50 million persons worldwide suffer from to study the bases of memory mechanisms on the
epilepsy [1], and more than forty clinical types of epi- nerve system of Aplysia [8], whereas Hodgkin and
lepsy have been described [2]. In spite on this diversity, Huxley succeeded with the study of the bases of nerve
the salient electrophysiological hallmark present in impulse on the squid’s giant axon [9].
most types of epileptic activity is the paroxysmal depo-
Several researchers studied the occurrence and
larization shift (PDS). During epileptic activity, some
spread of seizure activity in the buccal ganglia of the
neurons exhibit high-amplitude and long duration
depolarizations, these are the paroxysmal depolariza-
tion shifts of the membrane potential (see reference [3]
and Fig. 1).
Our knowledge about the neurophysiological bases
20 mV

of epilepsy are far from being complete, and there is

reason for hoping that understanding the biophysical
120 ms
bases of the PDS will clarify the pathophysiology of
epilepsy and perhaps delineate rational treatment
ways. In the context of the dominant ideas that regard
20 mV

epilepsy as the result of a misbalance between exci-

tation and inhibition in the nervous tissue [4–6], dif-
ferent authors suggest that the PDS is a gigantic post-
synaptic excitatory potential [7].
In neurophysiology it is customary to study any
phenomenon on the simplest biological object where
this phenomenon is manifested. This allowed Kandel
Acronyms: PDS—paroxysmal depolarization shift, KD—keto-
genic diet, PUFA—polyunsaturated fatty acids.
2 min
Fig. 1. Comparison of paroxysmal depolarizations
recorded intracellularly in cat’s cortical neurons (top), and
in a Helix B3 neuron (bottom). Upper part is adapted from
[3] (measurements were carried out under topical applica-
tion of pentylenetetrazole—PTZ). The lower part corre-
sponds to perfusion of 40 mM PTZ (unpublished data
from the authors).

956
 HYPOTHESIS ON POLLUTION OF NEURONAL MEMBRANES
Control
4 mM PTZ
8 mM PTZ
16 mM PTZ
40 mM PTZ
20 mV
1s
Fig. 2. Gradual transformation of the pacemaker potentials
into PDSs as pentylenetetrazole (PTZ) concentration
increases. The image was compiled on the basis of data
published in [15].
land snail Helix pomatia [7, 10–12]. Helix is endowed
with a relatively simple nervous system, and this allows
conducting biophysical experiments [13, 14]. In par-
ticular, each buccal ganglion of Helix have four giant
neurons, and from these, only neuron B3 systemati-
cally develops PDSs in the presence of pro-convulsive
drugs such as pentylenetetrazole (PTZ), etomidate,
heptanol (as well as low magnesium concentration,
etc.).
There is a long-standing discussion regarding the
similarities between the behavior of this single-cell
epilepsy model and the development of human epi-
lepsy, and today there is no doubt that from the view-
point of fundamental mechanisms involved, there is a
number of common features, notwithstanding the
large phylogenetic distance between humans and
snails [10].
The idea that pacemaker potentials are the basis of
epileptiform activity in the nervous system has been
BIOPHYSICS Vol. 66 No. 6 2021
957
discussed in literature [14]. The possibility to generate
PDSs in neurons completely isolated both from inver-
tebrates [10] and in vertebrates [11], as well as the fact
that the PDS is accompanied by an increase in the
impedance of neurons [7, 12] are not consistent with
the idea that PDSs have a synaptic origin. On the other
hand, in experiments with neuron B3 it was found ,
that with increasing doses of PTZ (and also etomi-
date) pacemaker potentials are gradually transformed,
increasing both duration and amplitude, to finally
convert into PDSs (see. Fig. 2).
The basic mechanisms of pacemaker potentials
generation still remain not completely clarified (see,
e.g., references [16, 17]). A “minimal” mathematical
model for the pacemaker potential in Helix pomatia
neurons has been proposed by Kononenko et al. [18–
22]. The model incorporates, firstly, two stationary
potential-independent conductances for sodium and
potassium, which conform a linear leakage current,
secondly, a sodium stationary potential-dependent
S-shaped conductance, which corresponds to the
region of negative resistance in the I-V trace, and
thirdly, a hyperpolarization-activated conductance.
Using the Kononenko model, mathematical modeling
was carried out and it was determined the set of bio-
physical parameters, which ensures the transition
from the pacemaker potential to the PDS [23]. It was
shown that PDS-like potentials may appear when
changing in model’s parameters, as well as through the
addition of noise at the input (see Fig. 3). At a noise
level of 0.2 arbitrary units (a. u.) the pacemaker poten-
tials lengthen and show a tendency to irregularity, at a
noise level of 0.6 a. u. a large and prolonged depolar-
ization, reminiscent of a PDS, appears (Figs. 3b, 3c)
[23].
THE MEMBRANE POLLUTION HYPOTHESIS
Ulrich Altrup (1943–2007), former head of the
Institute of Experimental Epilepsy Research at the
University of Münster (Germany), studied epilepti-
form activity induced at the buccal ganglia Helix for
almost 30 years. In the last years of his research activ-
ity, Altrup put forward the membrane pollution
hypothesis. Altrup believed that if substances that dif-
fer sharply in their mechanism of action, such as hep-
tanol, pentylenetetrazole, etomidate (as well as a low
concentration of magnesium), are capable of inducing
PDS, then the main mechanism of epileptogenesis
should be nonspecific.
To this it should be added that a variety of associ-
ated ionic mechanisms (such as leakage potassium
channels, a large range of ligand-gated channels, and
others) are influenced by the same range of concen-
trations of pentylenetetrazole (as well as etomidate).
These observations suggest the existence of a nonspe-
cific mechanism in epileptogenesis. Altrup and his
958 NECHIPURENKO et al.

50 (a) 7 6.8

ΔΠ, mN/m2
Vm, mV

4
3.2
3
1.9
2 1.5
–65 1
0 20 40 60 80 100 0.15
Time, s 0
(b) 1 mM 4 mM 8 mM 16 mM 40 mM
10
Pentylenetetrazole dose
0
–10 Fig. 4. Mean values of lateral pressure recorded at a Wil-
–20 helmy film balance corresponding to varying doses of pen-
tylenetetrazole. The image was compiled on the basis of
Vm, mV

–30 data published in reference [15].

–40
–50 A large amount of experimental evidences led
–60 Altrup to the idea that epileptogenic amphiphilic sub-
stances penetrate into the lipid bilayer membrane of
–70
nerve cells and increase the lateral pressure in the
0 50 100 150 200 250 300 bilayer, originating PDSs in those neurons that are
Time, s capable generating pacemaker potentials. Increased
10 (c) lateral pressure reduces membrane fluidity, influenc-
0 ing on the mobility of membrane proteins and causing
a transition from a liquid condensed state to a solid
–10
condensed state.
–20
Protein-lipid interactions and neuronal excitability.
Vm, mV

–30 Protein-lipid interactions and their relationship with

–40 excitability have been discussed for several decades in
–50 biophysical research [24]. Thus, authors of reference
[25] found that agents that alter membrane physico-
–60
chemical properties of (e.g., simple alcohols and ace-
–70 tone) are able to modify the affinity of Na, K-ATPase
0 100 200 300 400 500 600 to monovalent cations.
Time, s The most famous model for the generation and
propagation of a nerve impulse is the model of Hod-
Fig. 3. Gradual conversion of the pacemaker potential into gkin and Huxley [9]. Its biophysical plausibility as well
PDS as noise is added. Simulations were carried out using as its ability to explain many properties of nerve exci-
the Kononenko model [20]: (a) spontaneous pacemaker
potentials, obtained by the authors of [21]; (b) and (c) the tation led to the fact that this model has received uni-
result of adding white Gaussian noise (0.2 a. u. and versal recognition in neurosciences and cardiology.
0.6 a. u., respectively). A. Hodgkin and A. Huxley experimentally found that
membrane electric capacitance remains constant
during the time of excitation. Therefore, the role of
coworkers found in a Wilhelmy film balance, that the lipids in this model is overlooked, and any modifica-
epiletogenic substances pentylenetetrazole and etomi- tion of ion channels is considered as a result of inter-
date cause, in the same range of epileptogenic concen- actions of an electrostatic nature. In the early 1970’s,
trations, a gradual increase in lateral pressure lipid I. Tasaki proposed a theory, which does not fit in the
monolayers [7]. On Fig. 4 the results of these experi- Hodgkin-Huxley model: he assumed that the propa-
ments with the addition of pentylenetetrazole are illus- gation of nerve impulse is accompanied by a wave
trated. mechanical excitation along the axon [26]. These
ideas did not find further experimental confirmation;

BIOPHYSICS Vol. 66 No. 6 2021

 HYPOTHESIS ON POLLUTION OF NEURONAL MEMBRANES
at the same time, the registration of gating currents,
single-channels recordings as well as many other
observations testified in favor of the Hodgkin and
Huxley model [27].
Recently appeared evidences suggest that action
potential propagation is accompanied by a wave of
mechanical deformation of the cell membrane, which
moves at constant speed [28]. Various authors are cur-
rently attempting to construct a model capable of the-
oretically unifying all electrochemical, mechanical
and thermodynamic phenomena associated with the
generation and conduction of the nerve impulse. The
hypothesis of a mechanical soliton seems to us the
most attractive. However, there are still required a
number of experimental evidences and theoretical
developments for the idea of nervous impulse as a soli-
ton to be confirmed. In some studies [28–31] it was
suggested, that the opening of ion channels involved in
generation of nerve stimulation requires a mechanical
change in the lipid bilayer. In favor of this hypothesis
is the argument that during the activation of ion chan-
nels occurs a deformation of the membrane [28, 29].
On the other hand, enthalpy measurements during
action potential propagation do not correspond to the
predictions of Hodgkin and Huxley, but support the
mechanical impulse model [30, 31].
The authors of reference [31] claim that the action
potential, as modeled by Hodgkin and Huxley, reflects
the sum of all the potential changes over the entire
membrane, registered at a relatively large area, and in
reality it is the result of a combination of effects from
open channels, as well as closed and inactivated chan-
nels. Direct mechanical stimulation of the axon could
elicit the opening of sodium channels, in the course of
which is generated the energy required for maintaining
the movement of the mechanical soliton [32].
If these changes in mechanical properties of the
lipid bilayer accompanying the action potential will
intrinsically be related to the physiological process of
nerve activity, then it will be understandable why any
substance able to modify the physicochemical proper-
ties of the lipid bilayer may also modify the excitability
of neurons. Such an approach could provide a plausi-
ble explanation to the effect of inhaled anesthetics,
whose effectiveness correlates with their solubility in
lipids [33], and also the well-known antiepileptic
effect of a number of food additives, such as the
omega-3 polyunsaturated fatty acid [34], vitamin D
[35], and others [36]. We can also expect that the
change of membrane mechanical properties caused by
non-specific epileptogenic amphiphilic substances
also can affect pacemaker potentials, causing abnor-
mal changes that lead to epileptogenesis, in accor-
dance with the membrane pollution hypothesis [37].
The ketogenic diet and the membrane pollution
hypothesis. About 30% of patients with epilepsy are
resistant to drug therapy (more than 15 million people
BIOPHYSICS Vol. 66 No. 6 2021
959
worldwide). One of the therapeutic interventions that
can improve the condition of more than half of
patients with severe epilepsy is the ketogenic diet. Typ-
ically, the ketogenic diet (KD) is very rich in fat and
poor in carbohydrates (usually the proportion of fat to
non-fats is equal to four to one) and it has been used
for the treatment of refractory epilepsy. The value of
such a diet in a clinical context was noted in the 1920’s
in connection with its successful application for epi-
lepsy treatment.
The first reference stating that fasting may have an
antiepileptic action, appeared in 1911. Parisian doctors
J. Guelpa and A. Maria prescribed fasting for the
treatment of twenty patients and reported about allevi-
ating their symptoms [38]. In early 1920's, J. Conklin
suggested that epilepsy appears as result of “intoxica-
tion of the brain with substances coming from the
intestines”. Conklin administered to his patients a diet
consisting only of water during 30 days for the treat-
ment of refractory epilepsy and achieved success -
improving the condition in more than half of the
patients [39].
Representation that ketone bodies are the main
effector in the anticonvulsant effect of fasting led Rus-
sell Wilder in 1921 to the idea of formulating a diet
with a high content of fat and a low content of carbo-
hydrate for the treatment of epilepsy. Wilder obtained
encouraging results and called the diet ketogenic [40].
Beneficial effects of the diet were confirmed in the
1930’s, but then it ceased to be used by doctors, which
was connected to the appearance of phenytoin in 1938
and, later on, other antiepileptic drugs. However, for
the past 30 years, epilepsy refractory to pharmacother-
apy has attracted the attention of the scientific com-
munity, and again ketogenic diets are catching the
interest of specialists. Ketogenic diet remains as the
only dietary method for treating epilepsy; it has a sci-
entific rationale and has developed the principles for
its practical application as a therapy. The physiological
basis of KD is the establishment in the body of those
conditions which are similar to those that occur during
prolonged fasting. Among different variants of the KD
there is the classical KD, the diet enriched with tri-
glycerides of medium length, modified Atkins diet,
and low glycemic index diet [41–45].
Noteworthy, increased content of ketone bodies in
the organism is not necessarily connected an about
with diabetic ketoacidosis – a life-threatening condi-
tion. The concentration of ketone bodies in the blood
also increases as a result of the physiological response
to episodes of short-term or moderate fasting. The
goal of KD is to achieve ketosis while avoiding ketoac-
idosis. Thus, in a state of ketosis induced by KD, levels
of glucose, insulin and pH are kept close to normal
values, even when ketone bodies blood concentration
is almost two orders of magnitude higher than normal
values, but nevertheless this concentration remains
960 NECHIPURENKO et al.
three times lower than values characteristic for dia-
betic ketoacidosis (where glucose levels in blood is
increased by three times, the insulin concentration
falls almost to zero, and pH is lowered to 7.3, which is
incompatible with life [46]).
As result of the application of KD in 50–60% of
those patients who are resistant to several anticonvul-
sant drugs, a reduction by at least twice the frequency
of seizures is observed, and approximately 20% of
patients achieve their complete suppression. In addi-
tion, ketogenic diet can improve patient’s well-being
and cognitive performance [47]. The evidence for the
effectiveness of KD poses serious problems for theo-
retical epileptologists. In the following sections, we
will discuss the possible mechanisms of KD, consider-
ing their possible relationship with Altrup’s membrane
pollution hypothesis.
Metabolism under KD conditions. With prolonged
fasting or during diabetes mellitus, when the cell does
not have access to glucose, the body begins to utilize
fatty acids. Typically, this occurs via β-oxidation, and
a result acetyl-CoA is synthetized, which is incorpo-
rated into the Krebs cycle for generating ATP. How-
ever, with prolonged fasting, not all of the acetyl-CoA
can be metabolized through the Krebs cycle, and a sig-
nificant part of the acetyl-CoA is converted into
ketone bodies: acetoacetate and β-hydroxybutyrate, as
well as acetone (formed by non-enzymatic decarbox-
ylation of acetoacetate). Ketone bodies can provide up
to two thirds of the brain’s energy needs.
The ketogenic diet, impacting on overall energy
exchange, can generate shifts in metabolism, leading
to a reduced production of lactate and α-ketogluta-
rate. As a result, the concentration of glutamate
decreases and the risk of excitotoxicity is lowered.
Lactate is capable of inhibiting lipolysis [48]. The
increased level of lipolysis in the result of the reduced
levels of lactate with the KD, at the same time, lipids
and fatty acids included into the diet may provide cer-
tain changes in the membranes, contributing to the
mechanisms countering lipid bilayer pollution. The
limited availability of glutamate may play here an
important role. A reduction in glutamate levels leads to
an increase in the absorption of cysteine, and this
causes an increase in glutathione levels with the corre-
sponding removal of unnecessary amphiphilic sub-
stances that can contaminate the neuronal membrane.
Thus, with increased protection against oxidative
damage provided by glutathione, cells can avoid the
effect of membrane pollution during epileptogenesis
[49].
There is mounting evidence pointing to the idea
that KD exerts its anticonvulsant action through syn-
ergies, including several mechanisms. In this way,
antiepileptic effect of KD can find an explanation
beyond the traditional approach to the action on the
synaptic level, and many of the effects can be analyzed
from the viewpoint of the membrane pollution
hypothesis. In the context of Altrup’s hypothesis, we
will focus further on the following three aspects related
to KD:
(1) The anticonvulsant effect of ketone bodies;
(2) The connection of glutathione with KD antie-
pileptic effects;
(3) The contribution of polyunsaturated fatty acids
(PUFA) and cholesterol to the action of KD.
Anticonvulsant action of ketone bodies. For quite a
long time it has been discussed the hypothesis about
ketone bodies acting directly as pharmacological
agents. Kate in 1933 reported that seizures caused by
thujone in rabbits were prevented with previous treat-
ment with acetoacetate [50]. This was later confirmed
in a murine model of audiogenic seizures [51]. There
is also an early report on the anticonvulsant action of
acetone [52]. Authors of reference [53] reported a
dose-dependent suppression of convulsive seizures by
intra-peritoneal administration of acetone, in four
separate animal models of seizures and epilepsy [53].
In support of the idea that the effect of acetone is one
of the fundamental mechanisms of action KD, can be
added the fact that patients with epilepsy who posi-
tively responded to KD therapy, showed higher levels
of acetone in the brain (above 1 mmol/L) [54]. As
result authors of reference [53] put forward the idea
that acetone is the causative factor in KD anticonvul-
sant effects.
With regard to putative molecular targets for ketone
bodies it was found that VGLUT2 (a carrier of gluta-
mate into synaptic vesicles) is inhibited by acetoace-
tate [55] in the concentrations range expected for KD
treatment. This effect can mitigate the excitotoxicity
associated with glutamate. At the same time, ketone
bodies, apparently do not alter synaptic transmission
at the level of the hippocampus, which is in full accor-
dance with Altrup’s observations [7, 15]. Thus,
authors of reference [56], using methods of cellular
electrophysiology, found that millimolar concentra-
tions of β-hydroxybutyrate and acetoacetate do not
affect: (1) excitatory postsynaptic potentials and pop-
ulation discharges in CA1 pyramidal neurons after
stimulation of Schaffer’s collaterals; (2) spontaneous
epileptiform activity in the seizure model of the hippo-
campal-entorhinal cortex; and (3) ionic currents
induced by glutamate, kainate and γ-aminobutyric
acid in hippocampal neurons.
In particular, it has been identified a β-hydroxybu-
tyrate receptor, GPR109a (HCAR2), which is acti-
vated in mutant epileptic mice (mutUNG1) fed with
KD, which implies selective effect KD on mecha-
nisms of ketone bodies receptors [57, 58].
Other studies show that ketone bodies can also act
as neuro-protective agents [59, 60]. Thus, authors of
reference [61] have shown that β-hydroxybutyrate and
BIOPHYSICS Vol. 66 No. 6 2021
 HYPOTHESIS ON POLLUTION OF NEURONAL MEMBRANES
acetoacetate (in millimolar concentrations) prevent
hyperexcitability, caused by hydrogen peroxide in
acute neocortical slices and dissociated neurons, and
that ketone bodies are able to significantly reduce the
formation of active forms oxygen in isolated mito-
chondria. This indicates the effect of KD through the
activation of antioxidant mechanisms by fatty acids
and ketones. It was also shown that acetone is able to
increase the fluidity of the lipid membrane [62], act-
ing in the opposite direction of pro-convulsive sub-
stances, and this explains its antiepileptic effect in the
context of Altrup’s hypothesis. In general, it should be
summarized that among the many possible mecha-
nisms of the anticonvulsant action of ketone bodies,
their effect on membrane viscosity and the corre-
sponding stabilization of the lipid bilayer may be of
fundamental importance.
Glutathione and the mechanism of KD. A decrease
in glutamate levels as result of reduced glucose intake
leads to an increase in cysteine absorption, and this
causes an increase in glutathione (GSH) levels with
the corresponding removal of unnecessary amphi-
philic substances that can pollute the neuronal mem-
brane. Similarly, an excess of released glutamate in
synapses during seizures obstacles the absorption of
cysteine, a necessary component for glutathione syn-
thesis. Glutathione is a tripeptide thiol found practi-
cally in all cells; it is involved in redox reactions, and
protects the cell from reactive oxygen species (ROS)
[24–26]. Glutathione removes toxins and many alien
and toxic compounds via direct conjugation. In partic-
ular, due to the elimination of free radicals with gluta-
thione, the membrane structure is stabilized [33].
The role of glutathione in epilepsy is well known.
Thus in a pilocarpine seizure model in rats it was
found that the concentration of GSH decreased by 54
and 58% in the striatum and frontal cortex, respec-
tively, during convulsions [63]. In model experiments
on animals seizures it was observed the production of
large amounts of ROS, which lead to oxidative damage
to proteins and peroxide oxidation of lipids; this may
influence the activity of the antioxidant enzymes
superoxide dismutase and glutathione peroxidase.
Consequently, GSH levels in the brain decrease.
Thus, in [64] it was found a significant decrease
approximately to 35% of the average GSH/water ratio,
even in areas of the brain that remotely located respect
to the epileptic focus [65]. Thus, it is expected that
glutathione helps to reduce the level of membrane pol-
lution, obstructing the development of epilepsy [66].
Polyunsaturated fatty acids (PUFAs) and ketogenic
diet. A ketogenic diet enhances the mobilization of
PUFAs from adipose tissue to the liver and brain [57–
67]. One of the versions of this diet, the KD enriched
with medium- length triglycerides, is the method of
choice for drug-refractory epilepsy treatment. It was
found in a rat hippocampus slices pentylenetetrazole
BIOPHYSICS Vol. 66 No. 6 2021
961
seizures model that there is a branching-point specific
effect which is comparable to the antiepileptic effect of
valproic acid. This suggests that the fatty acids present
in the KD, may directly exert anticonvulsant effects
[68].
Another likely mechanism for the antiepileptic
effect of KD could be associated to cholesterol. In
1939 the authors of reference [69] asked themselves if
the mechanism of a ketogenic diet proceeds through
raising cholesterol levels. Studying the cocaine-
induced seizure model in rats, they found that intra-
peritoneal injection of cholesterol raises seizure
threshold. It was shown that ketogenic diet elicits a sig-
nificant and monotonic increase in the level of choles-
terol along a whole year of its application [70].
It is known that cholesterol regulates membrane
fluidity, and its antiepileptic effect may be associated
to this. Cholesterol contained in plasma membranes of
eukaryotes, modulates the fluidity of membranes,
making the membrane less liquid at a high tempera-
ture (e.g., body temperature at 37°C) at the expense of
limiting the mobility of phospholipids, and more liq-
uid at a low temperature at the expense of preventing
dense phospholipids packing [71].
Note that heptanol causes a decrease in the mem-
brane fluidity of cholesterol domains in the lipid
bilayer, and this effect could explain the epileptogenic
effect of heptanol [72]. In this respect is also worthy of
attention that propofol, which has anti-epileptic
action [73] is able to increase the fluidity of mem-
branes [74]. Thus, in contrast to other prevailing theo-
ries on epileptogenesis, the membrane contamination
hypothesis may provide a rational explanation for the
antiepileptic effect of KD and other experimentally
confirmed facts [68].
DISCUSSION
Anti-epileptic mechanism of the KD is still an
enigma for many researchers. Clinical and experimen-
tal evidences show that KD is effective even against the
most severe forms of epilepsy. Nevertheless, generally
accepted theories about the mechanisms of epilepto-
genesis are unable to explain this fact. We believe that
membrane pollution hypothesis can explain this
mechanism, and this is the main thesis from our work.
The causes for pollution of nerve cells membranes
in epileptogenesis may be diverse: brain tissue necrosis
( for example, due to trauma), toxins production in the
case of epilepsy induced by neuro-infection, or
amphiphilic substances release as result of inflamma-
tion or excitotoxicity.
In connection with the possible biophysical mech-
anisms of the anticonvulsant effect of KD, three main
points can be noted:
(1) Role of ketone bodies in the stabilization of the
lipid bilayer may be connected with their influence on
962 NECHIPURENKO et al.
Ketogenic diet
Ketone bodies Glutathione PUFA
increased increased increased
Membrane
decontamination
Antiepileptic effect
Fig. 5. Schematic representation of the mechanism of
action of ketogenic diet in connection with the membrane
pollution hypothesis. PUFA: polyunsaturated fatty acids.
such biophysical properties, as fluidity and also with
their antioxidant action.
(2) Increased glutathione activity promotes mem-
brane decontamination and prevents the action of oxi-
dizing agents.
(3) PUFA intake from a KD, may play an import-
ant role in stabilizing the damaged membrane in epi-
leptic brain tissue.
It is not excluded the contribution of other effects
in action of KD: diet can change the solvation lipids,
proteins and carbohydrates, as well as a number of
intermolecular interactions at the level of the mem-
brane, which may be disrupted during epilepsy and
restored as result of following a KD.
Note that there may be glial cells membrane pollu-
tion during epileptogenesis – glial cells do not develop
PDSs, but membrane pollution can lead to conse-
quences which aggravate the epileptic process. In par-
ticular, to the pollution of neuronal membranes with
glial metabolic by-products or to the synchronization
of the epileptic focus [15].
Generally, protein-lipid interactions play a key role
in the transition from the pacemaker potential to a
PDS where a set of proteins, ion channels, ion pumps,
receptors, etc. are participating. Thus, according to
Altrup, “epileptic activity may be considered to result
from a “pollution” of neuronal membranes with
amphiphilic substances…” Incorporation of these
substances should increase membrane pressure, and
by doing this, it may increase membrane resistance,
block voltage-sensitive K+-currents, block all types of
ligand-operated channels, and may transform pace-
maker potentials into PDSs. [7].
In Figure 5 a diagram is presented explaining anti-
convulsant effect of the KD in accordance with
Altrup’s hypothesis.
CONCLUSIONS
A collection of our as well as literature data offers
new facts supporting and refining the membrane pol-
lution hypothesis proposed by Ulrich Altrup. Keto-
genic diet can favorably modify the properties of the
lipid bilayer of the neuronal membrane through vari-
ous mechanisms:
— Direct action of acetone and other ketone bodies
(as anticonvulsant compounds).
— Enhancing the synthesis of cholesterol, which
stabilizes membrane fluidity and has antiepileptic
action.
— Reductions in excitotoxicity associated with glu-
tamate.
— Reductions in the levels of lactate, needed to
activate lipolysis.
— Raising the levels of glutathione, which reduces
the level of ROS and cleanses epileptogenic amphi-
philic substances
— Stabilizing action of PUFA.
ACKNOWLEDGMENTS
The authors consider as a pleasant duty to thank Yuri
Ermakov, Denis Semyonov and Leonid Yakovenko for use-
ful discussions, and I. Lavrinenko for technical assistance.
We are also deeply grateful to the referee for careful reading
our paper, positive feedback as well as interesting questions.
FINANCIAL SUPPORT
This work was performed under the support of the
Cuban Science Foundation (FONCI) - Project on non-
pharmacological antiepileptic therapy- coordinated by the
Cuban Center for Neurosciences, and also was supported by
the Program for Basic Research of State Academies of Sci-
ences of Russia for 2013–2020 (Topics nos. 01201363818
and 01201363820).
CONFLICT OF INTERESTS
Authors declare no conflict of interests.
ETHICAL ISSUES
This work does not include any description of research
performed on humans or animals.
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