RU2348421C2 - Brain cortex motoneurons growth stimulator for growing mammal and method of application thereof - Google Patents

Abstract

FIELD: medicine; experimental neurology.

SUBSTANCE: offered method of brain cortex motoneurons growth stimulation for growing mammals in experiment involves daily introduction of adrenal cortex extract within total course 5 weeks. Herewith pig's adrenal cortex extract is introduced in single dose 0.5 ml per 100 g of body weight. Offered is application of pig's adrenal cortex extract as brain cortex motoneurons growth stimulator for growing organism.

EFFECT: extended range of products for brain cortex motoneurons growth stimulation and possibility to use available product.

2 cl, 1 tbl

Description

The proposed group of inventions relates to experimental biology and medicine, namely to experimental neurology, and can be used to stimulate the growth of motor neurons in the cerebral cortex of a growing mammalian organism.

In the future (after conducting appropriate clinical studies), a possible area of use of the proposed group of inventions may be the prevention and treatment of cerebral palsy, since (as will be described below) an important pathogenetic factor in this disease is damage to the motor neurons of the cerebral cortex.

Cerebral palsy occupies a special place in the pathology of childhood, is the most common disease of the nervous system, leading to disability of children, affecting not only the motor system, but also causing impaired speech and intelligence (Semenova K.A.).

Cerebral palsy occurs as a result of brain damage transferred to the perinatal period of development or during the neonatal period. By now it has become clear that birth trauma, cerebral hemorrhage (which were previously considered the main cause of the disease) are mostly secondary phenomena and only in a relatively small number of cases they have independent significance in the etiology of cerebral palsy, resulting from various forms of obstetric pathology. The latter include not only the narrow pelvis of the mother or its incorrect structure, but also the incorrect position of the fetus. Under the influence of labor forces, deformation of the fetal head may occur with one or another presentation with a displacement of various parts of the brain and the possibility of the consequences of mechanical trauma in childbirth. Mechanical injury to the fetal head is usually accompanied by a violation of cerebral circulation, cerebral hemorrhage. Most often, hemorrhage occurs due to rupture of the sagittal or transverse sinuses, which in most cases leads to fetal death. With subarachnoid hemorrhage, as a result of rupture of small vessels of the brain and per diapedesin, deaths occur less frequently, and various forms of cerebral palsy often develop. Cerebral hemorrhage can occur not only as a result of mechanical rupture of blood vessels due to mechanical trauma in childbirth, but also asphyxiation that began in childbirth, chronic hypoxia associated with damage to the fetus before the onset of labor. It can be a consequence of many causes that affect the brain at different stages of development (embryogenesis and fetogenesis). Asphyxia in childbirth can only be the end result of lesions and developmental disorders. However, it remains unclear why, in some cases, a significant displacement of the brain (and therefore, sufficient severe trauma to it) does not cause any disturbances in the activity of the brain, while in others, slight deformation, which would seem to entail damage to it, can be considered as the cause of cerebral pathology, developing in the future. At present, there is no doubt that the main cause of cerebral palsy and secondary microcephaly in most cases is the severe condition of the fetus that occurs under the influence of any one or a complex of harmful factors during fetal development. Such harmful factors during pregnancy, as infectious agents, toxins, etc., can penetrate the placental barrier, affect the brain tissue of the fetus and lead to its destruction, delayed brain development.

A number of authors attach great importance in the pathogenesis of cerebral palsy to autoimmune conflict between mother and fetus, especially in severe forms. The beginning of this conflict occurs at different times of embryo and fetogenesis. The consequences of an autoimmune conflict can also affect postnatal ontogenesis, manifested by the presence of specific anti-brain antibodies (Semenova K.A. et al.). There are: 1) the early stage of cerebral palsy and 2) the initial and late residual stages of cerebral palsy. Accordingly, various underlying processes, clinical manifestations of the disease and therapeutic approaches are allowed.

On the first day after birth, silence is observed in the ward, there is no bright lighting, an elevated position is given in the heated bed, it is recommended to keep the cold at a distance of 2 cm from the baby’s head, moistened oxygen is periodically or constantly given.

Drug therapy is aimed at brain dehydration (40% glucose intravenously 5-10 ml, magnesium sulfate 25% 0.2 ml / kg body weight, native or dry plasma 10-15.0 ml intravenously, vikasol 0.002-0.003 g / day, anticonvulsants means, in case of violation of swallowing, feeding through a probe).

In the future, vitamin B12, glutamic acid, cerebrolysin, dibazole are prescribed to stimulate the development of the brain, and pyrogenal, midocal, gammalon, galantamine (or oxazyl), ATP are included in the treatment of the initial and late residual stages, and physiotherapeutic procedures, massage and gymnastics are widely used, styling for limbs, exercises for the development of speech.

With any damage (trauma, hemorrhage, infection), an inflammatory process develops, including in the brain tissues. The most powerful anti-inflammatory effects are corticosteroid hormones and synthetic drugs based on them.

Semenova K.A., Mastyukova E.M., Smuglin M.Ya (1972), Semenova K.A., Makhmudova N.M. (1979. P. 118) used corticosteroid drugs to treat cerebral palsy. In the complex of therapeutic measures for this purpose, immunogenesis inhibitors were used in the late neonatal period (after 10 days) - first, ACTH from 5 to 10 units for 1-2 months (Semenova K.A. 1972, p. 91), and then another group of children - cortin (representing an extract of the adrenal cortex of slaughter cattle), 0.3-0.5 ml intramuscularly every other day in combination with dexamethasone, 1/4 tab. (in a tablet of 0.5 mg) daily for a month (Semenova K.A. et al. 1983). It was noted that in children who did not receive cortin and dexamethasone (group 1 - 10 people), positive dynamics of psychomotor development was observed in 45.0% of cases, while those treated with cortin and dexamethasone (group 2 - 20 people) in 54.0% of cases; certain positive changes were observed in the clinical picture, namely: manifestations of hypertension syndrome decreased slightly, installation reflexes appeared, muscle tone decreased, which was high before treatment, manipulative movements of the hands appeared. Positive changes in the immunological status were also noted and the representation of urinary corticosteroids improved.

Corticosteroids are widely used in the prenatal and postnatal period to reduce respiratory distress at the risk of premature birth of newborns who underwent intrauterine and childbirth hypoxia, had intracranial hemorrhage, and hemodynamic disorders (not controlled by dopamine) due to congenital weakness of the adrenal glands in preterm children. For this purpose, corticosteroids were used for the most part in a single short cycle (hydrocortisone). Repeated courses were recommended only with prolonged intubation and persistent hypotension. As an alternative strategy, steroids for inhalation, methylprednisolone, and hydrocortisone (rather than dexamethasone) are recommended.

At the same time, along with positive results (a decrease in the frequency of premature births, an increase in the survival rate of small children), attention is drawn to an increase in the incidence of ulcerative enterocolitis, sepsis in children in such cases, fears are expressed for the consequences, including the possibility of cerebral palsy and delayed neural mental development. It should be noted that corticosteroid drugs, having a suppressive effect on immunocompetent cells, can significantly affect the neurogenesis of a growing organism, especially in utero and in the neonatal period (Rajadurai V.S., Tan K.N., 2003).

Over the past decades, it has become generally accepted that structural neuroplasticity is involved in the functional adaptation of the central nervous system. Brain cell morphology is in continuous transition throughout life span as a response to environmental stimuli. The effects are to some extent associated with a changing level of gonodal steroid hormones. Today it is clear that the functions of gonadal steroids in the brain extend beyond the simple regulation of reproductive or neuroendocrine events. It is important that gonadal steroids are involved in the formation of the developing brain and throughout life are involved in higher brain functions such as mood, cognition and memory (Bovenberg S.A. et al, 2005). Gonadally induced changes in the neuron were established: the magnitude, number of synapses, as well as tree and synaptic morphology (Parducz, 2006).

In recent years, interest has increased in the proliferation of new neurons from precursors in the pancreatic structures of the brain (dentate gyrus and hippocampus), their migration to other regions, which are more pronounced at an early age, but, decreasing, persist throughout life.

Adult mammalian dentate gyros contain nerve cell precursors with multipotential development and self-renewal. The origin and maturation of nerve precursors are determined in accordance with the composition and level of trophic factors in their microenvironment (Kosaka N. et al, 2006). Trophic factors: ciliary neurotrophic factor (CNTF), glial-derived neurotrophic factor (GDNF), insulin-like growth factors 1 and 2 (IGF-1, IGF-2): the most active of them in promoting neurogenesis, ciliary factor (CMTT) causes rapid neuron enlargement (Chen H b, 2006).

The role of sex hormones of predominantly adrenal origin in neurogenesis has been shown experimentally on brain tissue cultures. Enzyme systems have been found that provide synthesis from precursors (pregnenolone, progesterone, dehydroepianrosterone) of testosterone neurohormones and estrogens in brain tissues. Estrogen alpha and beta receptors significantly increase hippocampal neurogenesis; stimulation is accompanied by a rapid increase in cells (Massucco, 2006).

As in other tissues, neurogenesis is modulated by growth factors, in which the transforming growth factor beta-1 (TGF-beta-1), which is synthesized by astropites, glia cells, plays an important role. (Galbiati M. Et al, 2005; Chen H. Et al, 2006). Hippocampal astrocytes synthesize dehydroepiandrosterone (3 times more than cortical astrocytes), as well as testosterone and estrogens from pregnenolone with the participation of cytochrome p450c17 (Zwain, 1999).

Beta growth TGF - inhibits hippocampal neurogenesis of growing mice (Buckwalter, 2006), and dehydroepiandrosterone reduces the activity of the transforming growth factor beta - 1, which inhibits the formation of neurons from precursors in cultures. Dehydroepiandrosterone stimulates the oxidative mechanism of energy production in the brain mitochondria of developing rats, significantly increases ATPase activity, dehydrogenase levels, accelerates the maturation of brain mitochondria, and thereby emphasizes the role of dehydroepiandrosterone in brain development in postpartum life (Patel, 2006).

The anti-inflammatory properties of dehydroepiandrosterone, which can lower the levels of proinflammatory cytokines - interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) Leowattana W, 2001, are noted. Glucocorticosteroids inhibit the release of gonadotropin hormone (GnRH), acting not only on the hypoph but directly at the hypothalamic level (Calogero AE et al. 1999).

The inhibitory effect of corticosterone on the release of GnRH was not modeled by androgens (T uDHT), but was eliminated by progesterone, at least when corticosterone was used in low concentrations (10 nitrometers), but not high (100 nitromethanes); in rats, the effects of corticosterone are consistent with those of hydrocortisone in humans.

At the same time, the suppressive effects of high concentrations of glucocorticosteroids on the division of neuronal precursors, their differentiation, and apaptosis were noted. By altering the metabolism of hippocampal neurons, glucocorticoids increase the vulnerability of the hippocampus to accumulating toxic products (Robert M Sapolsky, 1985; 2003). Low and medium concentrations of corticosteroids in hippocampal cell culture have anti-inflammatory effects (expression of IL-1beta and TNF-alfa decreases), while at high concentrations the effects were opposite, pro-inflammatory, concentrations of IL-1beta, TNF-alfa increased (Mac Pherson A et al., 2005).

Dehydroepiandrosterone (DHEA), in contrast, protects hippocampal neurons; antiglucocorticoid actions of DHEA are carried out through a decrease in glucocorticoid hormone receptors (Cardounel A. et al.)

Depending on the intensity, prenatal stress (stress) increases or inhibits the development of hippocampal neurons: short duration, moderate prenatal stress increased neurogenesis and differentiation of hippocampal neuron processes, while prolonged, serious stress harmed their morphology. Mineralocorticoid and glucocorticoid receptors contribute to stress-induced morphological changes in hippocampal elements; the mineralocorticoid receptor increased neurogenesis and differentiation in the culture of hippocampal neurons; in contrast, the glucocorticoid receptor has been implicated in suppressing their morphology (Fujioka A et al. 2006).

Seizures in the first weeks of life of newborns lead to a decrease in neurogenesis and later differentiation of neurons (Shy X. Y et al., 2006).

In low concentrations, hydrocortisone and progesterone enhance the effects of growth factors (such as insulin-like growth factor, etc.). In connection with the foregoing, an increase in neurogenesis can be expected when using adrenal cortex extracts containing a complex of hormone-active compounds that have hormonal and paracrine effects on metabolic processes in the brain. The possibility of using extracts of pork and fetal adrenal cortex deserves special consideration, since at the fetal age, this gland, together with the placenta, constituting a single hormonal complex, ensures rapid growth and differentiation of fetal tissues.

Thus, the methods currently used in the clinic for the prevention and treatment of cerebral palsy are not directly aimed at stimulating cerebral cortical motor neurons. The applied methods do not provide a sufficient effect and have a large number of undesirable side effects.

In experimental medicine, there are also practically no developments related to stimulating the growth of cerebral cortical motor neurons. This direction is only beginning to develop. In the well-known literature, we have found only one work devoted to a means for stimulating the growth of cerebral cortical motor neurons and the method of its application. We selected this work as a prototype - I.G. Shilenok, I.V. Mukhina, I.V.Sadovnikova, N.A. Panin. Anabolic effects of minor concentrations of corticosteroids contained in adrenal cortex extracts. // Nizhny Novgorod Medical Journal. - 2004. - No. 1, p. 92-95. In the prototype, to stimulate the growth of cerebral cortical motor neurons (in an experiment on rats), it was proposed to use an extract of the fetal adrenal cortex in a volume of 0.5 ml every other day for 5 weeks. Despite the obtained growth effect of motor neurons in the cerebral cortex, the prototype is not without drawbacks. First of all, it should be noted that fetal adrenal glands were taken at the autopsy of children who died in childbirth. Obviously, the use of the prototype in this regard is very limited, primarily ethical issues.

The objective of the invention is to expand the arsenal of tools to stimulate the growth of motor neurons in the cerebral cortex, providing the possibility of using an affordable tool.

The task is achieved by using the extract of the pork adrenal cortex as a means to stimulate the growth of motor neurons in the cerebral cortex of a growing mammalian organism.

The objective of the method of stimulating the growth of motor neurons of the cerebral cortex of a growing mammalian organism, including the administration of an adrenal cortex extract, every other day, in a general course of 5 weeks, is achieved by administering an extract of pork adrenal cortex in a single dose of 0.5 ml per 100 g of body weight .

The possibility of using extracts of pork adrenal cortex as a means to stimulate the growth of motor neurons in the cerebral cortex of a growing organism is shown for the first time in the present invention. Earlier extracts of pork adrenal cortex were used for completely different purposes, namely: to stimulate reparative processes in a pathologically altered liver (Ru 2185835 C2, 07.27.2002); for the correction of acute renal failure in toxic hepatorenal syndrome (Ru 2290940 C2, 10.01.2007).

In the present invention, the stimulation of the growth of cerebral cortical motor neurons is carried out precisely on a growing organism, since it was intended to extrapolate the results for use in the treatment of pediatric patients.

An essential feature is the quantitative indicators entered into the claims, namely, a single dose and a course of administration of extracts of pork adrenal glands. A single dose of 0.5 ml per 100 g of mass was determined by the appropriateness of testing the minimum (minor) concentrations of hormone-active substances that are usually found in the blood without the stress of the body and have a biological effect (not in milligrams and micrograms, but in nanograms). The need to use a 5-week course is explained by the gradual development of the effect and therefore the required long stimulation time, which is usually used in clinical conditions in the treatment of patients, courses for 4-6 weeks.

The need for the introduction of extracts every other day is explained by the fact that with subcutaneous administration, the drug is slowly resorbed and thereby a greater uniformity of concentrations can be achieved over a long period.

The proposed method was developed in an experiment on 25 growing outbred male rats with a body weight of 99.8 ± 3.7 g at the beginning of the experiment and a body length of 12.44 ± 0.58 cm. At the end of the experiment, the animals had a weight of 181.7 ± 12. 7 g and a body length of 14.0 ± 1.66 cm. The animals were divided into 5 groups, of which 5 animals were intact (1 control group), 5 animals received a 10% solution of ethanol in physiological sodium chloride solution (2 control group ), 5 animals received gadrocortisone (10 mg / kg) based on a 10% solution of ethyl alcohol (group 3); 5 animals received an extract of the pork adrenal cortex (group 4) and 5 animals received an extract of the bark of fetal adrenal glands taken at autopsy of human fetuses who died in childbirth (group 5).

Adrenal cortex extracts were prepared according to the principle of organ preparations; 1 ml of extract was obtained from 3.0 g of tissue. Preparations in a volume of 0.5 ml were administered every other day for 5 weeks. The method of radioimmunological analysis in extracts of the adrenal cortex determined the concentrations of hydrocortisone, dehydroepiandrosterone and progesterone. Hydrocortisone — 65 nmol, dehydroepiandrosterone — 0.83 nmol, and progesterone — 1.1 nmol, were contained in 0.5 ml of pork adrenal cortex extract. In 0.5 ml of extract of the fetal adrenal cortex contained hydrocortisone - 5.0 nmol, dehydroepiandrosterone - 1.15 nmol and progesterone - 0.45 nmol. In 0.5 ml of the hydrocortisone solution administered to the animals, 2.6 μmol of the preparation was contained. After 16 injections of the preparations, the animals were killed, the brain was paraffinized, and the sections were stained using the Nisslet method. Microscopic measurements were carried out in 4-5 layers of the cortex at a magnification of 7 × 90, and two diameters (large and small) of the body and nucleus of motoneuron cells were determined using a scale. The value of the scale division was determined using a micrometer; one scale division was 16.6 mkm. When calculating the area used the formula

Smkm2 = 3.14 * 0.12 ² * (a / 2 * 16.6) * (b / 2 * 16.6), where 0.12 ² is the correction factor for the optical medium of the microscope, a is the large diameter, b - small diameter, 16.6 - the value of one division of the scale. The diameters of the body and cell nucleus were determined by the formula:

d1, d2mkm = 3.14 * 0.12 * a (or b) * 16.6. The sizes of motor neurons thus calculated were comparable with the normative data cited by Yu.M. Zhabotinsky in the monograph Normal and Pathological Morphology of the Neuron, 1965.

Studies are presented in the table.

As studies have shown, in group 2 of the control with the introduction of 10% alcohol on an isotonic sodium chloride solution, the obtained data did not differ from the corresponding measurements of group 1 of the control in intact animals or were insignificant. Of the features of the effect of alcohol on brain neurons, a slight increase in the small diameter of the neuron body (28.77 ± 4.63 μm versus 24.41 ± 3.08 μm) draws attention, which determined the rounding of the cells and a slight increase in the neuron's body area. However, with the same core parameters in both control groups (in intact and with the introduction of ethanol solution), the s2 / s1 ratio turned out to be close (0.2 versus 0.23), which does not allow us to think about a significant increase in the metabolic activity of the neuron.

With the introduction of hydrocortisone, the parameters of the body of the neuron and its nucleus (small and large diameters, body and nucleus areas) were close to those of intact animals, although their difference in the size of the nuclei could be statistically confirmed (p = 0.002; 0.017; 0.000). The ratio of the core and body areas corresponded to the data for intact ones (1.92).

Significant and one direction were changes in brain neurons after administration of extracts of the pork and fetal adrenal cortex. All parameters of the body and nucleus of neurons were significantly greater than that of intact animals and those receiving hydrocortisone injections. Of the features, greater rounding of the body and nucleus of the cell after administration of the pig adrenal gland extract (d2 / d1 of the body = 1.54 and nucleus = 1.54) was noted, which may reflect a higher activation of neurons and, possibly, is associated with a higher concentration of progesterone in the extracts of pork adrenal glands (1.1 nmol versus 0.45 nmol in a dose). With the introduction of extracts of fetal adrenal glands, the ratio of large and small body diameters was closer to the normative data (1.63 versus 1.54; in intact 1.93). The nucleus had a similar direction of changes in large and small diameters (1.77 versus 1.54; in intact 1.65) due to a smaller increase in small diameter (with an equal increase in large diameter, which determines cell growth), which suggests a greater effect of extracts fetal adrenal glands on neuron growth in length, and is possibly associated with a high content of dehydroepiandrosterone (1.15 nmol per injection versus 0.83 nmol in pork adrenal gland extracts) and 13 times less hydrocortisone (5 nmol versus 65 nmol).

It should be noted that in previous studies in animals, it was found that ATP was highly secured in brain tissue under conditions of hypobaric hypoxia, both with the addition of extracts of the fetal adrenal cortex and extracts of the pork adrenal cortex. In comparison with the control groups and after the administration of hydrocortisone, the differences were highly significant. After administration of extracts of fetal adrenal glands, the concentration of ATP and the ratio of ATP / AMP were slightly better than after administration of extract of pork adrenal glands (2.489 ± 0.109 compared to 2.305 ± 0.134; respectively, the coefficient. ATP / AMP was 10.637 ± 0.463 versus 8.801 ± 1.777). However, in the hypoxic control group, the ATP concentration was 1.732 ± 0.216, and the ATP / AMP ratio was only 4.977 ± 1.265. All this suggests that the extracts of pork adrenal cortex are not inferior to the extracts of the fetal adrenal cortex in stimulating effects. Given the fact that raw materials for obtaining extracts of pork adrenal glands are higher than for extracts of fetal adrenal cortex, the possibility of using the former is more promising.

It should be noted that examinations of experimental animals in the dynamics of the experiment showed that the largest increase in weight over a 5-week period was observed in intact animals (90 g); animals that were injected with extracts of pork and fetal adrenal glands added 82.4 g and 84.0 g, respectively (in the group with hydrocortisone - 80.2 g).

It is noteworthy that the increase in growth in the intact control group was 1.26 cm; the highest increase was observed in animals that were injected with extracts of pork (2.0 cm) and fetal adrenal glands (2.46 cm), less - with the introduction of hydrocortisone (1.46 cm).

Considering that in animals of these groups large sizes of bodies and nuclei of neurons were also observed, we attribute the above changes in the neurons of growing animals not so much to manifestations of regulatory hypertrophy as to true growth due to all cell structures.

Literature

Semenova K.A., Mastyukova E.M. Smuglin M.Ya. Clinic and rehabilitation therapy for cerebral palsy. Medicine-Moscow 1972.

Semenova K.A., Zhukovskaya E.D., Yakovleva N.T., Semenov A.S., Dukarsky F.G. The effect of glucocorticosteroid drugs on immunological parameters and the functional state of the adrenal collar in young children with cerebral palsy. Pediatrics. 1983, 12, 25-27.

Bovenberg S.A., van Uum S.H., Hermus A.R. Dehydroepiandrosterone administration in humans: evidence based? Neth J. Med. 2005, 63 (8): 300-4.

Buckwalter M.S., Yamane M., Coleman B.S. Onnerod B.K., Chin J.T., Wyss-Coray T. Chronically increased transforming growth factor-beta 1 strongly inhibits hippocampal neurogenesis in aged mice. Am J. Pathol, 2006; 169 (1): 154-64.

Calogero A.E., Burello N., Bosboom A.M., Garofalo M.R., Weber R.F., D Agata R.

Glucocorticoids inhibt gonadotropin-releassing hormones by acting directly at the hypothalamic level. Endocrinol. Invest., 1999; 22 (9): 666-70.

Cardounel A., Regelson W., Kalimi M. Dehydroepiandrosterone protecs hippocampal neurons against neurotoxin-induced cell death: mechanism of action, Proc. Soc / Exp. Biol., 1999; 222 (2): 145-9.

Chen H., Tung Y.C., Li B., Iqbal K., Grundke-Iqbal I. Trophic factors counteract FGF-2-induced inhibition of adult neurogenesis. Neurobiol Aging., 2006.19 [Epub ahead of print] Cristi L. Watterberg M.D. Jeffrey S. Gerdes M.D. Xathleen L. Gitibrd R.N. and Hung Mo Lin Ph D Prophylaxis Againts Early Adrenal Insuffiency to Prevent Chronic Lung Disease in Premature Infants / Pediatrics vol 104 No 6, 1999, 1258-1263

Fujioka A., Fujioka T., Ishida Y., Mackawa T., Nakamura S. Differential effects jf prenatal stress on the morphological maturation of hippocampal neurons. Neuroscience, 2006, 25; 141 (2): 907-15.

Galbiati M., Saredi S., Romano N., Martim L., Motta M., Melcangi R.C. Smad proteins are targets of transforming grouth factor betal in immortalized gonadotrophin-releasing hormone releasing neurones. J. Neuroendocrinol. 2005; 17 (11): 753-60.

Gregory A. Lodigensky M.D., Karin Reademaver M.D., Sewa Zimine Ph.D., Marianne Groenendaal M.D. Rhd. Linda S de Vrics M.D. Ph D and Petra S Huppi M.D. Structural and Functional Brain Development Affer Hydrocortisone Treatment for Neonatal chronic Lung. Pediatrics vol 116, No. 1, 2005, 1-7.

Rajadurai V.S., Tan K.H. The use abuse of steroids in perinatal medicine. Ann Acad Med Singapore. 2003; 32 (3): 324-34.

Kosaka N., Kodama M., Sasaki H., Yamamoto Y., Takeshita F., Takahama Y., Sakamoto H., Kato T, Terada M., Ochiya T. FGF-4 regulates neural progenitor cell proliferation and neurosal differentiation. Faseb J., 2006; 20 (9): 1484-5.

Leowattana W. DHEA (S): the fountainof youth. J Med. Assoc. Thai 2001; 84 Suppi 2: S605-12 MacPherson A., Dinkel K., Sapolsky R. Glucocorticoids worsen excitotoxin-induced expression of pro-inflammatory cytokines in hippocampal cultures. Exp Neurol. 2005; 194 (2): 376-83.

Mazzucco C.A., Lieblich S.E., Bingham B.I., Wi] liamson M.A., Viau V., Galea L.A. Both estrogen receptor alpha and estrogen receptor beta agonists enhance cell proliferation in the dentate gyros of adult female rats. Neuroscience. 2006, 15; 141 (4): 1793-800.

Patel M.A., Katyare S.S. Dehydroepiandrosterone (DHEA) treatment stimulates oxidative energy metabolism in the cerebral mitochondria from developing rats. Int. J. Dev Neurosci. 2006; 24 (5): 327-34.

Robert M. Sapolsky. Glucocorticoid toxicity in the Hippocampus: Temporal aspects of neuronal vulnerability. Bram Research, v 359.1-2.19985: 300-305.

Shi X.Y., Wang J.W., Sun R.P. Effects and consequence of recurrent seizures of neonatal rat on the hippocampal neurogenesis. Zhonghua Er Ke Za Zhi. 2006; 44 (4): 289-93.

Zwain I H., Yen S.S. Dehydroepiandrosterone: biosynthesis and metabolism in the brain. Endocrinology, 1999; 140 (2): 880-7.

Claims (2)

1. The use of pork adrenal cortex extract as a means to stimulate the growth of motor neurons in the cerebral cortex of a growing mammalian organism. 2. A method of stimulating the growth of motor neurons of the cerebral cortex of a growing mammalian organism in an experiment, comprising administering an adrenal cortex extract every other day, in a general course of 5 weeks, characterized in that a single dose of pork adrenal cortex extract is administered in a single dose of 0.5 ml per 100 g of body weight .