RU2410107C1 - Method of preventing and correction of disturbances of main functions of central nervous system in case of lead intoxication in prenatal and early postpartum period - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: in order to prevent and correct central nervous system (CNS) disorders in case of lead intoxication in prenatal and early postpartum period preparation baliz-2 is introduced in doses 0.3 ml per 1 kg of body weight during all pregnancy for 7 days before delivery and for 7 days after delivery.

EFFECT: efficient protection of brain against lead intoxication due to reduction of free-radical oxidation intensity and increase of cerebellum neurons survival rate.

3 tbl, 3 ex

Description

The invention relates to medicine, namely to experimental pharmacology and toxicology, and for the prevention and correction of disorders of the central nervous system (CNS) during lead intoxication in the prenatal and early postpartum period in high-risk areas.

Recently, there has been an increasing pollution of the environment by xenobiotics and their increasing entry into the human body. Extremely important is the effect on the body of heavy metals. This threatens the health and even life of all living things, including humans, as it damages cells and causes mutations leading to malignant processes or hereditary diseases (Kulinsky V.I. Xenobiotic neutralization // Soros Educational Journal. 1999. No. 1. P. 8. -12).

Lead is one of the most toxic and dangerous heavy metals. The source of significant emissions of lead into the atmosphere is mainly antiknock alkyl lead additives to fuel. Combined-cycle and aerosol lead compounds pose a great danger to the body. Studies by many authors have shown that lead can have a number of toxic effects at very low levels of exposure. Acute and chronic effects on human health can have neurological, cardiovascular, renal, gastrointestinal, hematological and reproductive effects. The central nervous system is especially susceptible to the harmful effects of lead. The toxic effect of this metal is manifested in the destruction of the blood-brain barrier, which leads to edema, loss of neurons and glia (Canfield RL, Henderson CR Jr., Cory-Slechta DA, Cox C, Jusko TA, and Lanphear BP Intellectual impairment in children with blood lead concentration bewol 10 microg per deciliter // N. Engl. J. Med. - 2003 .-- Vol. 348. - 1517-1526).

At the same time, the developing brain is especially prone to the harmful effects of lead. Prenatal and postpartum action of lead affects the higher functions of the central nervous system: it inhibits brain development, distorts behavioral reactions (motor disinhibition, excessive irritability and irritability) (Ryzhavsky B.Ya., Mikhailov V.I., Feldsherov Yu.I. et al. Effect of introduction lead to pregnant rats on the brain of their offspring (long-term consequences) // Bull.exp. biol. and honey. - 2000. - T.129. - No. 1. - P.28-30; Privalova LI, Kuzmin S .V., Malykh OL and others. The role of environmental pollution by lead in delaying the psychological development of children preschool age // Bulletin of the Russian Academy of Medical Sciences. - 2002. - No. 11. - S.50-53; Chukhlovina M. L. Lead and nervous system (review) // Gig. and dignity. - 1977. - No. 5. - S. 39-42; Strazyńska L., Dabrovska-Bonta LB, Koza K. and Sulkowski G. Inflammation-Like Glial Response in Lead-Exposed Immature Rat Brain // Toxicololotical sciences. - 2007. - Vol. 95. - N1 . - P.1-26).

A number of studies have shown that disorders in learning, attention, memory appear even at a blood lead content of 10 μg / dl, and according to some reports even at 5 μg / dl (Toscano Chr.D., Guilarte TR Lead neurotoxicity: from exposure to molecular effects // Brain research reviews. - 2005. - Vol. 49. - P.529-554; USEPA, Lead: Identification of dangerous levels of lead, in: CFR. - 2001. - Vol.40. - p. Part 745). Studies of cognitive function have shown that increasing blood lead for every 10 μg / dL leads to a decrease in IQ by 0-5 points (Bellinger D. Lead and neuropsychological function in children: progress and problems in establishing brain-behavior relationships // Adv Child Neuropsychol. - 1995. - Vol. 3. - P.12-45). According to Zerbino et al, small doses of lead with prolonged action are of particular danger (Zerbino D.D., Solomenchuk T.I., Pospishil Yu.A. Lead - an etiological factor in vascular damage: the main evidence // Pathology archive. - 1997. - T.59. - No. 1. - S.9-11). Neurotoxicity of low levels of Pb with prolonged exposure is especially evident in children. Numerous data show a direct relationship between exposure to low doses of lead and behavioral and cognitive impairment in childhood (Finkelstein Y., Markowitz M.E., Rosen JF Low-level lead-induced neurotoxicity in children: an update on central nervous system effects // Brain Research Reviews. 1998. Vol. 27. P.168-176).

The occurrence of pathological conditions during lead poisoning, according to many authors, is associated with the activation of free radical oxidation processes (Kundiev Yu.I., Stegka VA, Dmitruha NN and others. Dependence of changes in the immune and biochemical mechanisms of maintaining homeostasis on material cumulation lead in the body (experimental study) // Occupational medicine and industrial ecology. - 2001. - No. 5. - P.11-17; Adonaylo VN, Oteiza PI Lead intoxication: antioxidant defenses and oxidative damage in rat brain // Toxicology. - 1999. - Vol. 135. - P.77-85; Bokara KK et al. Lead-induced increase in antioxidant enzymes and lipid peroxidation products in developing rat brain // Biometals. - 2008. Vol.21. - P.9-16).

The enhancement of lipid peroxidation has a membrane-toxic effect, which leads to cell disintegration, while various enzyme systems undergo significant damage [Mkhitaryan VG, Agadzhanov MI, Gevorkyan DM and other Enzymatic mechanisms of anti-radical protection of cells in extreme conditions // Vestnik AMN. - 1982. - No. 9. - S.15-19].

One of the important links in protecting the cell from the toxic effects of hydroperoxide is the antioxidant glutathione system [Zozulya Yu.A., Baraboy V.A., Sutkova D.A. Free radical oxidation and antioxidant protection in brain pathology. M. - 2000. - 344 p .; Kulinsky V.I. Neutralization of Xenobiotics // Soros Educational Journal. 1999. No. 1. S.8-1].

Currently known methods of preventing dysfunctions of the central nervous system during prolonged exposure of lead using exogenous antioxidants [Patrick L. // Altern. Med.Rev. 2006. Vol.11, N 2. P.114-127]. However, modern pharmacological agents intended for the treatment of acute and chronic poisoning with salts of heavy metals have a number of negative effects, which significantly limits their use [Rogan W.J. Safety efficacy of succimer toddlers with blood lead level of 22-24 µg / dL // Pediatr. Res. 2000. Vol. 48. N.5. P.593-599].

Curcumin is known to be used to protect the brain in lead intoxication. Its parallel use with lead led to an increase in the content of reduced glutathione, a decrease in lipid peroxidation, and significantly normalized the activity of superoxide dismutase and catalase in such regions of the brain as the frontal cortex, striatum, hippocampus, cerebellum [Shukla PK, Khanna VK, Khan MY Srimal RC Protective effect of curcumin against lead neurotoxicity in rat // Human & Experimental Toxicology. 2003. Vol.22. P.653-658]. Curcumin is a polyphenol. It is insoluble in water, but readily soluble in alcohol and ether.

The use of tea catechins is known for exposure to HepG2 human cells in culture. The positive effect was manifested in increased cell survival, decreased lipid peroxidation and membrane fluidity [Chen L., Yang X., Jiao h., Zhao B. Tea Catechins protect against lead-indused cytotoxocity, lipidperoxidation and membrane fluidity in HepG2 cells // Toxicological Science. 2002. Vol. 69. P. 149-156].

However, catechins in high concentrations or in the presence of Cu ²⁺ induce DNA damage and increase the oxidation of unsaturated fatty acids, manifesting themselves as prooxidants [Hayakawa F., Kimura T., Maeda T. et al. DNA cleavage reaction and linoleic acid peroxidation induced by tea catechins in the presence of cupric ion // Biochim. Biophys. Acta. 1997. Vol. 1336. P. 123-131; Shen SR, Yang XQ, Zhao BL Prooxidant effect of tea polyphenols in vitro // J. Tea Science. 1992. Vol. 12. P.145-150].

Closest to the claimed is a method for the prevention and correction of disorders of the main functions of the central nervous system during prenatal lead intoxication using the drug "Histochrome", by parenteral administration at a dose of 0.1 mg / kg to pregnant females a day before lead nitrate intoxication at a dose of 200 mg / kg through a gastric tube and the day after intoxication. An experimental study of the drug "Histochrome", as an antioxidant in lead intoxication in the early period of ontogenesis, showed that double intraperitoneal use of it at a dose of 1 mg / kg a day before and, a day after the use of lead nitrate in pregnant female rats on 17-18 days of pregnancy at a dose of 200 mg / kg in the brain of their offspring reduces the long-term effects of toxic effects of lead, affecting the development of oxidative stress, suppressing it [Ryzhavsky B.Ya., Lebedko OA, Belolyubskaya DS The effect of the drug "Histochrome" on the severity of the long-term effects of the prenatal effects of lead nitrate in the brain of rats. // Bull. an experiment. biol. and honey. 2008. Volume 146. No. 8. S.236-240]. The drug "Histochrome" is used in cardiology and ophthalmology.

The disadvantages of the method for the prevention and correction of central nervous system disorders with the Histochrome drug in case of lead intoxication include contraindications for its use during pregnancy and lactation, as well as in childhood and with hypersensitivity to the components of the drug [http://amt.allergist.ru/gistohrom_b. html, http://amt.allergist.ru/product/gistohrom.html, Directory of medicines, homeopathy, dietary supplement]. A side effect of the drug is moderate pain after the injection, as well as allergic reactions.

The technical task is to develop a method for the prevention and correction of disorders of the main functions of the central nervous system during lead intoxication in the prenatal and early postpartum period.

To solve the technical problem, it is proposed in the method for the prevention and correction of disorders in the developing central nervous system during lead intoxication in the prenatal and early postpartum period to experimentally administer the drug baliz-2 to female rats throughout pregnancy and in the early postpartum period per os, once a day, on an empty stomach at a dose of 0.3 ml / kg body weight. It is possible to use the drug Baliz-2 in female rats in the last 7 days of pregnancy and in the early postpartum period for 7 days at a dose of 0.3 ml / kg body weight.

The drug baliz-2 has anti-inflammatory, antimicrobial and regenerative properties. It is used in medicine for the treatment of wounds of various etiologies, burns, ulcers of the gastrointestinal tract, has antistress, antioxidant and antimutagenic effects. Baliz-2 is not toxic, does not have teratogenic or embryotoxic properties, does not have a mutagenic effect on bone marrow cells of experimental animals [M.D. Mashkovsky. Medicines A guide to pharmacotherapy for doctors. T.2. Ed. 10, Sr., M: "Medicine". - 1986. - 576 s].

To solve the technical problem, the following groups of female rats were formed:

1. intact (pregnant females who did not receive drugs);

2. receiving lead acetate throughout pregnancy and the early postpartum period;

3. received lead acetate + baliz-2 - 0.1 ml / kg throughout pregnancy and in the early postpartum period for 7 days;

4. receiving lead acetate + baliz-2 - 0.3 ml / kg throughout pregnancy and in the early postpartum period for 7 days;

5. receiving lead acetate + baliz-2 - 0.5 ml / kg throughout pregnancy and in the early postpartum period for 7 days;

6. received lead acetate + baliz-2 - 0.1 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days;

7. receiving lead acetate + baliz-2 - 0.3 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days;

8. receiving baliz-2 in the last 7 days of pregnancy and in the early postpartum period for 7 days at a dose of 0.5 ml / kg;

9. receiving baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.1 ml / kg;

10. receiving baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.3 ml / kg;

11. receiving baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.5 ml / kg;

12. receiving baliz-2 - 0.1 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days;

13. receiving baliz-2 - 0.3 ml / kg of the last 7 days of pregnancy and in the early postpartum period for 7 days;

14. receiving baliz-2 - 0.5 ml / kg during the last 7 days of pregnancy and in the early postpartum period for 7 days.

Lead intoxication was modeled by introducing lead acetate along with food to female Wistar rats weighing 200-250 g and 5.0-5.5 months old at a dose of 6 mg / kg body weight daily, throughout pregnancy and in the early postpartum period (in within 7 days after delivery).

The effect of chronic lead intoxication on the central nervous system of offspring was evaluated in 7-day-old rats born from female rats in the formed groups according to indicators such as free radical oxidation in the brain of offspring, the state of antioxidant glutathione protection in the brain, and the survival of cerebellar neurons in culture with glutamate neurotoxicity.

The level of free radical oxidation was determined by the chemiluminescent method (Farkhutdinov UR, Farkhutdinov PP // Bull. Expert biol. And medical. - 2000. - T.129. - No. 3. - S.260-264) on the device SmartLum 5773. The light sum of chemiluminescence was evaluated. The experimental results were determined by the severity of chemiluminescence (in cu). Statistical analysis of the results was carried out using Student's criterion [Lakin G.F. Biometrics. M., 1990. 352 pp.] In the editor of Microsoft Office Exel 2003, data are presented as M ± m.

Example 1

The experiment was conducted on the offspring (126 rats from 25 females) of rats, 9 rats in each group, and after decapitation, free radical oxidation (CPO) was determined in the brain. The results are presented in table 1.

Table 1 The effect of balise-2 on free radical oxidation in the brain of rat pups when modeling chronic lead intoxication (M ± m) No. p / p Groups of animals Light sum (c.u.) one Intact 144.07 ± 1.88 2 Lead Acetate 180.68 ± 2.53 ^∗∗∗ 3 Received lead acetate + baliz-2 - 0.1 ml / kg throughout pregnancy and in the early postpartum period for 7 days 172 ± 2.18 ^∗∗∗ four Receiving lead acetate + baliz-2 - 0.3 ml / kg throughout pregnancy and in the early postpartum period for 7 days 155.26 ± 2.89 ^∗∗∗ _∗∗∗ 5 Receiving lead acetate + baliz-2 - 0.5 ml / kg throughout pregnancy and in the early postpartum period for 7 days 159.16 ± 3.12 ^∗∗∗ _∗∗ 6 Those receiving lead acetate + baliz-2 - 0.1 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days 169 ± 2.28 ^∗∗ _∗ 7 Receiving lead acetate + baliz-2 - 0.3 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days 156.58 ± 3.92 ^∗ _∗∗∗ 8 Received lead acetate + baliz-2 in the last 7 days of pregnancy and in the early postpartum period for 7 days at a dose of 0.5 ml / kg 157 ± 3.11 ^∗∗ _∗∗ 9 Received baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.1 ml / kg 159 ± 2,13 ^∗∗ _∗∗ 10 Received baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.3 ml / kg 155.35 ± 3.24 ^∗∗ _∗∗∗ eleven Received baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.5 ml / kg 154 ± 2.58 ^∗ _∗∗ 12 Received baliz-2 - 0.1 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days 160 ± 1.48 ^∗∗∗ _∗∗ 13 Received baliz-2 - 0.3 ml / kg of the last 7 days of pregnancy and in the early postpartum period for 7 days 154.86 ± 2.48 ^∗∗ _∗∗∗ fourteen Received baliz-2 - 0.5 ml / kg of the last 7 days of pregnancy and in the early postpartum period for 7 days 154.05 ± 1.93 ^∗∗ _∗∗∗ Note: ^∗ - p≤0.05; ^∗∗ p≤0.01; ∗∗∗ p≤0.001 compared with intact control; * - p≤0.05; _∗∗ p≤0.01; _∗∗∗ p≤0.001 compared with data for lead intoxication.

The table shows that in the brain of 7-day old rat pups treated with lead acetate throughout the prenatal and early postpartum period of development (group 2), the level of free radical oxidation (CPO) was significantly higher than in rat pups born from intact females (group No. 1). The use of balise-2 in doses of 0.1; 0.3 and 0.5 ml / kg of weight during the whole pregnancy of females and the early postpartum period (groups 9, 10, 11) did not practically change the level of free radical oxidation in the brain of rat pups. At the same time, the intensity of CPO remained at the level of intact control. There were no changes in the level of CPO in the brain of rat pups in groups 12, 13, 14 (receiving baliz-2 in the same doses during the last 7 days of pregnancy and in the early postpartum period). In the brain of rat pups whose mothers received during the prenatal and early postpartum period along with lead acetate Baliz-2 throughout pregnancy and in the early postpartum period for 7 days at a dose of 0.1 ml / kg body weight (group No. 3), the level of free radical oxidation, although it was slightly lower than in rat pups poisoned with lead acetate, but significantly higher than that in the brain of intact rat pups. The most pronounced and almost identical antioxidant effect of baliza-2 was observed with the use of baliza-2 against the background of lead intoxication in doses of 0.3 and 0.5 ml / kg of body weight of a pregnant female (groups No. 4 and No. 5). Similar results were found in the brain of rat pups, whose mothers received lead acetate throughout pregnancy and the early postpartum period (7 days after childbirth) and received baliz-2 in doses in the last 7 days of pregnancy and the early postpartum period (7 days after childbirth) 0.1; 0.3 and 0.5 ml / kg of mass (groups No. 6, No. 7, No. 8).

Thus, it was found that the use of balise-2 at doses of 0.3 and 0.5 ml / kg against the background of prenatal and early postpartum lead intoxication has an enhanced and almost identical antioxidant effect. In this regard, further studies of the antioxidant properties of baliza-2 (its effect on antioxidant glutathione protection) were carried out on rats born to females who received balis-2 at a dose of 0.3 ml / kg of the body against the background of lead intoxication.

Example 2

To study the effect of baliza-2 on antioxidant glutathione protection and the level of MDA in the brain of weekly rat pups who received lead acetate in the prenatal and early postpartum period, the activity of glutathione peroxidase enzymes was studied [Moin V.M. A simple and specific method for determining the activity of glutathione peroxidase in red blood cells // Laboratory. - 1986. - No. 12. - S.724-727] and glutathione reductase [Yusupova LB On improving the accuracy of determining the activity of erythrocyte glutathione reductase. // Laboratory work. - 1989. - No. 4. - S.100-101], the content of oxidized and reduced glutathione [Thannhauser T.W., Konishi Y., Scheraga H.A. Sensitive quantitative analysis of disulfide Bonds in polypeptides and proteins // Analytical biochemistry. - 1984. - Vol. 138. - P.181-188], as well as the accumulation of one of the secondary products of lipid peroxidation of malondialdehyde (MDA) [Gavrilov VB, Gavrilova AR, Mazhul L.M. Determination of the content of lipid peroxidation products in the test with thiobarbituric acid. // Questions honey. chemistry. - 1987.-№1. - S.19-21].

Statistical analysis of the results was carried out using Student's criterion [Lakin G.F. Biometrics. M., 1990. 352 pp.] In the editor of Microsoft Office Exel 2003, data are presented as M ± m.

The experiment was carried out on 90 (from 18 females) 7-day-old Wistar rats whose mothers throughout pregnancy and in the early postpartum period (7 days after birth) received a dose of 0.3 ml / kg of balis-2 in the presence of lead intoxication body weight, as well as on rats whose mothers throughout the pregnancy received daily lead acetate at a dose of 6 mg / kg body weight daily and 7 days before birth and within 7 days after birth, the drug baliz-2 at a dose 0.3 ml / kg body weight. The following groups of female rats were formed:

1. intact (pregnant females who did not receive drugs);

2. receiving lead acetate throughout pregnancy and the early postpartum period;

3. receiving lead acetate + baliz-2 - 0.3 ml / kg throughout pregnancy and in the early postpartum period for 7 days;

4. receiving lead acetate + baliz-2 - 0.3 ml / kg in the last 7 days of pregnancy and in the early postpartum period for 7 days;

5. receiving baliz-2 throughout pregnancy and in the early postpartum period at a dose of 0.3 ml / kg;

6. receiving baliz-2 - 0.3 ml / kg for the last 7 days of pregnancy and in the early postpartum period for 7 days;

The research results are shown in table 2.

table 2 The effect of baliza-2 on antioxidant glutathione protection and the level of MDA in the brain of weekly rat pups receiving lead acetate in the prenatal and early postpartum period. No. p / p Groups of animals Glutathione reduced (μM / 1 g protein) Glutathione oxidized (μM / 1 g protein) Glutathione peroxidase (μM reduced glut. / 1 min 1 g protein) Glutathione reductase (μM NADP / 1c 1 g protein) MDA (nM / 1 g protein) one Intact 101.28 ± 3.12 11.75 ± 0.40 480.69 ± 17.82 0.87 ± 0.02 5.19 ± 0.18 2 Lead Acetate 95.83 ± 2.16 17.51 ± 0.58 ^∗∗∗ 590.20 ± 12.72 ^∗∗∗ 0.78 ± 0.01 ^∗∗∗ 7.14 ± 0.21 ^∗∗∗ 3 Lead Acetate + Baliz-2 throughout pregnancy and the early postpartum period 104.49 ± 2.23 _∗∗ 12.48 ± 0.41 _∗∗∗ 493.77 ± 13.96 _∗∗∗ 0.83 ± 0.02 _∗ 4.97 ± 0.25 _∗∗∗ four Lead Acetate + Baliz-2 in the last 7 days of pregnancy and in the early postpartum period 109.56 ± 2.64 _∗∗∗ 12.50 ± 1.06 _∗∗∗ 514.5 ± 22.94 _∗∗∗ 0.78 ± 0.01 _∗∗∗ 5.67 ± 0.13 ^∗ _∗∗∗ 5 Baliz-2 in
flow
throughout
pregnancy and the early postpartum period 109.57 ± 2.51 ^∗ _∗∗∗ 12.31 ± 0.68 _∗∗∗ 409.00 ± 25.02 ^∗ _∗∗∗ 0.91 ± 0.04 ^∗∗ 4.7 ± 0.17 ^∗ _∗∗∗ 6 Baliz-2 in the last 7 days of pregnancy and in the early postpartum period 103.27 ± 2.17 ^∗ _∗∗∗ 12.56 ± 1.17 ^∗ _∗∗∗ 442 ± 13.19 ^∗ _∗∗∗ 0.81 ± 0.07 ^∗ _∗∗∗ 4.92 ± 0.09 ^∗ _∗∗∗ Note: ^∗ - p≤0.05; ^∗∗ p≤0.01; ^∗∗∗ p≤0.001 compared with intact control;
_∗ p <0.05; _∗∗ p≤0.01; _∗∗∗ p≤0.001 compared with data of lead intoxication.

Analysis of the data presented in the table shows that in group No. 2 there is a significant decrease in comparison with group No. 1 - intact rats, antioxidant glutathione protection in the brain of 7-day old rats. In this case, a decrease in the content of reduced glutathione, an increase in the content of oxidized glutathione, an increase in the activity of glutathione peroxidase and a decrease in the activity of glutathione reductase are observed. In the brain of these animals, a significant increase in the content of one of the secondary products of lipid peroxidation, malondialdehyde, was observed in comparison with group No. 1.

In the brain of rat pups of groups No. 5 and No. 6, on the contrary, a small but significant increase in the activity of antioxidant glutathione protection was detected. So, in comparison with group No. 1, the content of reduced glutathione increased, the activity of glutathione reductase slightly increased and the activity of glutathione peroxidase decreased, the content of MDA significantly decreased.

The use of baliza-2 in group No. 3 contributed to the restoration of the activity of antioxidant glutathione protection to its level in group No. 1. Moreover, all the indicators of glutathione metabolism that we studied in the brain of these rat pups did not practically differ from those in the intact control (group No. 1). The restoration of antioxidant glutathione protection was also noted with the use of baliza-2 in group No. 4. In groups No. 3 and No. 4, there was also a decrease to the level of intact control of the content of the secondary product of MDA lipid peroxidation (Table 2).

Thus, chronic lead intoxication of rat pups in the prenatal and early postpartum period at a dose of 6 mg / kg body weight of a pregnant female is accompanied by a pronounced increase in free radical oxidation in the brain of 7-day-old rat pups, in a decrease in antioxidant glutathione protection, and contributes to the increased accumulation of one of the lipid peroxidation products MDA. The use of baliza-2 at a dose of 0.3 ml / kg body weight of a pregnant female against the background of lead intoxication during the entire period of pregnancy and in the early postpartum period, and in the last 7 days of pregnancy and 7 days after birth significantly reduces the intensity of free radical oxidation, helps to restore the activity of the antioxidant glutathione system.

Example 3

The effect of baliza-2 on the survival of brain neurons during lead intoxication in the prenatal and early postpartum period was studied in a culture of cereal granular cells under conditions of glutamate neurotoxicity [Viktorov IV, Khaspekov LG, Shashkova N.A. // Guide to the cultivation of nerve tissue: methods, techniques, problems. - M .: Science. - 1986. - S.141-166] ..

The studies were performed on a 7-8-day-old cerebellum granule cell culture obtained from the brain of 7-9-day-old Wistar rat pups, intact and born to females throughout pregnancy and in the postpartum period of 7-8 days, treated with 6 mg lead acetate / kg body weight.

Cerebellar neuron cultures obtained from intact rat pups and rat pups prenatally and in the early postpartum period of lead acetate etched were divided into the following groups, respectively:

1.1., 2.1 control - cultivated in incubation saline solution (ISR);

1.2., 2.2. ISR + baliz-2 - 50 μl / ml;

3.1., 3.2. ISR with glutamate (Glu) 100 μm;

4.1, 4.2. ISR with glutamate (Glu) 100 μM + 50 μl / ml baliza-2.

Statistical analysis of the results was carried out in the editor of Microsoft Office Excel 2003 using t-student criterion. The results of the study are presented in table 3

Table 3 The effect of baliza-2 on the survival of cerebellar neurons after exposure to glutamate on the background of prenatal and early postpartum intoxication No. p / p cultural groups % surviving neurons in culture groups after exposure: 1. control 2. Baliz-2 3. (Glu) 100 μM 4. (Glu) 100 μM + 50 μl / ml balise-2. one. cultures from intact rat pups 86.76 ± 1.32 85.82 ± 3.30 15.19 ± 3.06 35.22 ± 5.32 ^∗∗ 2. cultures from rat pups prenatally and in the early postpartum period of lead poisoned 84.79 ± 1.32 86.57 ± 0.99 7.41 ± 1.50 ^∗ 30.17 ± 5.02 ^∗ _∗∗ Note: ^∗ - p <0.05; ^∗∗ - p <0.001 - in comparison with the glutamate group for cultures from intact rat pups; _∗∗ - p <0.001 - in comparison with the group of glutamate for cultures of rats subjected to lead intoxication.

The analysis of the presented data shows that in the culture of granular cells of the cerebellum of rat pups (group 2.1) prenatally inoculated with lead acetate, the survival of neurons is practically the same as the intact culture (group 1.1). Ten minutes of exposure to glutamate causes the death of most of the cells in the culture of neurons obtained from the brain of intact rat pups (group No. 1.3) and even more pronounced death of neurons in the culture obtained from rat pups prenatally inoculated with lead acetate (group No. 2.3). The introduction of baliza-2 into the culture of neurons of these rat pups after exposure to glutamate significantly increases the survival of neurons. So when using Baliza-2, the survival of cerebellar neurons after exposure to glutamate in group No. 1.4 (cultures from intact rat pups) was 20% more than in group No. 1.3. Higher (by 22.8%) cerebellum granule cell survival after exposure to glutamate and balize-2 was also observed in the culture of neurons from the cerebellum of rat pups prenatally inoculated with lead acetate (group 2.4.).

An analysis of the above data indicates that prenatal chronic lead intoxication significantly enhances the neurotoxicity of glutamate. The use of baliza-2 in the culture of cerebellar neurons against the background of lead intoxication contributes to a marked decrease in the toxicity of glutamate.

Thus, we found that chronic lead intoxication of rat pups in the prenatal and early postpartum period at a dose of 6 mg of lead acetate per 1 kg of body weight of a pregnant female is characterized by pronounced oxidative stress in the brain of 7-day-old offspring, which is manifested by a significant increase in free radical oxidation in the brain 7 -day rat pups (Table 1, group. No. 2), in reducing the antioxidant glutathione protection, contributes to the enhanced accumulation of one of the products of lipid peroxidation - MDA (Table 2, group No. 2), which significantly enhances glutamate neurotoxicity (table 3, group No. 2.1). The use of baliza-2 at a dose of 0.3 ml / kg body weight of a pregnant female against lead intoxication during the entire period of pregnancy and in the early postpartum period, and in the last 7 days before childbirth and 7 days after birth significantly reduces the intensity of free radical oxidation (Table 1, groups No. 4, No. 7), helps to restore the activity of the antioxidant glutathione system (Table 2, groups No. 3, No. 4). It was also found that the use of the drug baliz-2 at a dose of 50 μl after the toxic effects of glutamate against the background of lead intoxication contributes to a significant increase in the survival of cerebellar neurons in culture (Table 3, group 2.4.).

Claims (1)

A method of protecting the brain of experimental animals from lead intoxication in the prenatal and early postpartum period, including dosing the drug, characterized in that the drug used is baliz-2 in doses of 0.3 ml per 1 kg of body weight throughout pregnancy or within 7 days before delivery and 7 days after delivery.