RU2590976C2 - Method of preparing antimutagenic product - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine and toxicology, in particular, therapeutic nutrition of population in zones of ecological trouble. Disclosed is method of preparing antimutagenic product in form of Marmalade mixture of gooseberry juice ordinary (L.Ribes uva-crispa) and girasol tubers (L.Helianthus tuberosus), including production by means of pressing diffusion of gooseberry ordinary and girasol juices, concentration of juice by evaporation to reduce volume in 2 times, mixing of concentrated juices in volume ratio of 1:1 and evaporation during 3-4 hours in a water bath until gelation or dry substances content is 68-70 %.

EFFECT: invention provides anti-mutagenic Marmalade product based on shared vegetal raw materials allows protection of genome of wide range of population, including children.

1 cl, 3 ex, 1 tbl

Description

The invention relates to the field of medicine and toxicology, in particular, therapeutic and prophylactic nutrition of the population in areas of ecological distress, and can be used to reduce the genotoxic load on the body cells caused by cadmium.

A known method of antimutagenic protection of the genome with an aqueous extract of meadow clover in a therapeutic dose of 0.002 g (dry matter) per 100 g of body weight.

The disadvantage is the use of wild plant raw materials of the local flora of Alpine meadows (Chopikashvili L.V. The cytogenetic effect of lead acetate and cadmium iodide in the Dr. melanogaster test system against therapeutic doses of dioxidine and the cytostatic preparation of cyclophosphamide and its correction with the infusion of clover meadow Trifolium pratense (local flora alpine meadows) / L.V. Chopikashvili, E.G. Pukhaeva, F.G. Tedeeva and others // Vestnik MANEB. - 2011. - V. 16, No. 2 (additional issue) 34-39 s) .

A known method of antimutagenic effects on the body of the drug "Melaxen" in doses of 5.25 · 10 ^-7 mg to 21.0 · 10 ^-7 mg per 100 g of live weight, daily for 3-7 days before bedtime against a background of cadmium Cd · 10 ^-7 M (patent No. 2440135 from 01.20.2012).

The disadvantage is that melaxen is a chemical analogue of the biogenic amine melatonin, which has side effects, according to the instructions, such as morning drowsiness, headache, etc.

Closest to the claimed solution is a method for producing an antimutagenic phytococktail, including the root system of eleutherococcus plants, licorice and Rhodiola rosea in 70% alcohol solution and the additional introduction of an 8-10% alcohol solution of green mass of meadow clover, while the phytococktail is taken according to 40 drops for 15-16 days daily (patent No. 2381809 of 02.20.2010).

The disadvantage of this method is the multicomponent use of expensive plant materials, the use of ethyl alcohol.

The objective of the invention is to obtain an antimutagenic gel-like product in the form of marmalade based on commonly available plant materials, which allows the protection of the genome of a wide range of people, including children.

The technical solution is achieved by the fact that in the method for producing an antimutagenic product, which includes administering antimutagenic substances of plant origin in predetermined doses, in contrast to the prototype, a jelly-like product is used in the form of marmalade from a mixture of juices of gooseberry berries (Ribes uva-crispa L.) and Jerusalem artichoke tubers ( Helianthus tuberosus L.) in a 1: 1 ratio, in doses of 4 to 6 g per 1 kg of live weight, daily for 7 days against a background of Cd · 10 ^-7 Μ.

A rich set of essential amino acids, a high content of inulin, dietary fiber, minerals, vitamins (B ₁ , B ₂ , B ₃ , B ₅ , B ₆ , B ₇ , C, PP, carotene, choline), organic acids (citric, malic , fumaric, amber, hinny) make Jerusalem artichoke a valuable raw material for the production of food products for medical purposes and extremely useful for children, vegetarians and the elderly

Gooseberries among berry shrubs in terms of vitamins C, P, B ₁ , Br, A, PP and trace elements K, Ca, Mg, Ρ, Fe are not inferior even to currants. Gooseberries contain hypotensive and capillary-strengthening, anti-sclerotic P-active compounds (bioflavonoids) with antimutagenic activity. Gooseberry berries contain a large amount of ascorbic and folic acid, which help strengthen blood vessels and enhance hematopoietic function. Therefore, the juice of gooseberry berries is widely used for vitamin deficiency, anemia, hemorrhage and skin diseases.

Due to the high content of pectins, which belong to the class of food antimutagens, Jerusalem artichoke and gooseberries have the ability to bind heavy metals and radionuclides in the human intestines, further removing them from the body. The main properties of pectin substances, which determine their application in the food industry, are gel-forming and complex-forming ability. Pectins in a jelly-like state are much more stable and are not subject to destruction. Therefore, the juices of gooseberries and Jerusalem artichoke tubers in a modified gelatinous state can enhance their phytotherapeutic effect.

The method is as follows.

The berries of gooseberry ordinary (Ribes uva-crispa L.) and Jerusalem artichoke tubers (Helianthus tuberosus L.) are subjected to preliminary cleaning and washing, then they are crushed and separately obtained by pressing the diffusion juice of berries and vegetables. Before blending, the juices are concentrated, evaporated to reduce the volume by 2 times, then mixed in a 1: 1 ratio, followed by evaporation for 3-4 hours in a water bath until gelation or solids content of 68-70%. The semi-finished product is poured into molds and cooled at room temperature for 10-12 hours. Thus obtained antimutagenic product in the form of marmalade is freed from molds and Packed in plastic bags.

The effectiveness of this antimutagenic product was confirmed by a series of experiments lasting from 3 to 10 days against a background of Cd · 10 ⁷ M.

In the experiments, 55 animals (rats (Rattus) of the Wistar line) at 11 points of the experiment were used:

1. Negative control - dry briquetted feed (7 days).

2. Positive control - CdI ₂ 10 ml / kg, based on 1 kg of animal body weight, at a concentration of 1 · 10 ^-7 Μ (3 days).

3. Antimutagenic product 2-4 g / kg, based on 1 kg of animal body weight (7 days).

4. Antimutagenic product 2-4 g / kg, (7 days) + CdI ₂ 10 ^-7 M 10 ml / kg (3 days).

5. CdI ₂ 10 ^-7 M 10 ml / kg (3 days) + Antimutagenic product 2-4 g / kg (7 days).

6. Antimutagenic product 4-6 g / kg, based on 1 kg of animal body weight (7 days).

7. Antimutagenic product 4-6 g / kg, (7 days) + CdI ₂ 10 ^-7 M 10 ml / kg (3 days).

8. CdI ₂ 10 ^-7 M 10 ml / kg (3 days) + Antimutagenic product 4-6 g / kg (7 days).

9. Antimutagenic product 6-8 g / kg, based on 1 kg of animal body weight (7 days).

10. Antimutagenic product 6-8 g / kg, (7 days) + CdI ₂ 10 ^-7 M 10 ml / kg (3 days).

11. CdI ₂ 10 ^-7 M 10 ml / kg (3 days) + Antimutagenic product 6-8 g / kg (7 days).

To evaluate the effectiveness of the proposed method for producing an antimutagenic product using genetic research methods, we studied the cytogenetic parameters in the cells of the bone marrow, lungs, bladder and large intestine in mammals (Wistar rats). We used such methods of cytogenetic analysis as:

- metaphase analysis in mammalian bone marrow cells (rats (Rattus) of the Wistar line);

- multiorgan micronuclear test on the organs of the gastrointestinal tract, lungs, urogenital system (rats (Rattus) of the Wistar line).

The characteristic antimutagenic activity of the obtained product (marmalade) containing juices of Jerusalem artichoke tubers (Helianthus tuberosus L.) and common gooseberries (Ribes uva-crispa L.) was characterized by mutagenesis induced by cadmium iodide according to the following indicators:

- the level of chromosomal aberrations (HA) in bone marrow cells;

- the level of abnormalities of the nucleus (AL) in the lung cells (pneumocytes of type I, type II and alveolar macrophages in total were taken into account), and epithelium of the bladder and large intestine.

Example 1

The berries of gooseberry ordinary (Ribes uva-crispa L.) and Jerusalem artichoke tubers (Helianthus tuberosus L.) are subjected to preliminary cleaning and washing, then they are crushed and separately obtained by pressing the diffusion juice of berries and vegetables. Before blending, the juices are concentrated, evaporated to reduce the volume by 2 times, then mixed in a 1: 1 ratio, followed by evaporation for 3-4 hours in a water bath until gelation or solids content of 68-70%. The semi-finished product is poured into molds and cooled at room temperature for 10-12 hours. Thus obtained antimutagenic product in the form of marmalade is freed from molds and Packed in plastic bags.

In this series of experiments, we studied an antimutagenic product based on the juice of Jerusalem artichoke tubers and gooseberries 2-4 g / kg (based on 1 kg of the animal’s body weight), against the background of negative and positive controls, in variants of pre- and post-treatment with model mutagen - cadmium iodide . An analysis of the experimental results presented in Table 1 showed that cadmium exhibits mutagenic properties both in bone marrow cells and in cells of the lungs, bladder, and colon. The number of chromosomal aberrations induced by cadmium iodide (option 2) compared with the negative control (option 1) increased 5 times, and the level of nuclear abnormalities in the cells of the lungs and bladder increased almost 2 times, the colon in 2.5 times.

Indicators of the level of chromosomal aberrations when using an antimutagenic product at a dose of 2-4 g / kg (option 3) are statistically close to the negative control (option 1), and against the background of the positive control (option 2), they decreased by 6 times. In the pre- and post-treatment with an antimutagenic product at a dose of 2-4 g / kg against cadmium (options 3 and 4), the level of chromosomal aberrations decreased to 2.8 and 3.4%, respectively, compared with the positive control (option 2).

The level of abnormalities of the nucleus of the lung cells in the positive control (option 2) was 7.19%, in the negative control (option 1) 4.05 ppm, and the use of the antimutagenic product reduced the rate to 3.02%. In the variants of pre- and post-treatment with an antimutagenic product against a background of CdI ₂ 10 ^-7 M, the level of core anomalies decreased to 5.26 and 4.75 ppm.

The level of abnormalities of the nucleus of the bladder cells in the positive control (option 2) was 8.62 ‰, in the negative control (option 1) was 4.05 ppm. The use of an antimutagenic product (option 3) reduced the level of nuclear abnormalities by half against the background of spontaneous mutagenesis and four times against the background of cadmium induced. In the variants of pre- and post-treatment with an antimutagenic product against a background of CdI ₂ 10 ^-7 M, the level of core anomalies decreased, respectively, to 2.97 and 3.58 ppm.

The level of abnormalities of the nucleus of the colon cells in the positive control (option 2) was 4.75%, in the negative control (option 1) 1.86 ppm. The level of abnormalities of the nucleus when using an antimutagenic product (option 3) is statistically close to the negative control (option 1) and four times less than the positive control (option 2). In the variants of pre- and post-treatment with an antimutagenic product against a background of CdI ₂ 10 ^-7 M, the level of core anomalies decreased to 1.77 and 2.97 ppm, respectively.

Thus, the antimutagenic product based on the juice of Jerusalem artichoke tubers and gooseberries has a modifying effect, reducing the clastogenic effect of cadmium in the cells of the bone marrow, lungs, bladder and colon.

Example 2

The berries of gooseberry ordinary (Ribes uva-crispa L.) and Jerusalem artichoke tubers (Helianthus tuberosus L.) are subjected to preliminary cleaning and washing, then they are crushed and separately obtained by pressing the diffusion juice of berries and vegetables. Before blending, the juices are concentrated, evaporated to reduce the volume by 2 times, then mixed in a 1: 1 ratio, followed by evaporation for 3-4 hours in a water bath until gelation or solids content of 68-70%. The semi-finished product is poured into molds and cooled at room temperature for 10-12 hours. Thus obtained antimutagenic product in the form of marmalade is freed from molds and Packed in plastic bags.

In this series of experiments, we studied an antimutagenic product based on the juice of Jerusalem artichoke tubers and gooseberry berries 4-6 g / kg (based on 1 kg of the animal’s body weight), against the background of negative and positive controls, in the variants of pre- and post-treatment with model mutagen - cadmium iodide .

Analysis of the experimental results presented in table 1 showed that the antimutagenic effect in options 6, 7, 8 is higher than in options 3. 4, 5. Thus, the antimutagenic product has a positive modifying effect at a dose of 4-6 g / kg (options 6 ) against the background of negative control (option 1), reducing the level of chromosomal aberrations of spontaneous mutagenesis by 3 times. The level of abnormalities of the nucleus of the bladder and colon is statistically close to the indicator of negative control (option 1). The antimutagenic product (options 3) exhibits a modifying effect against the background of negative control (option 1), reducing the level of nuclear abnormalities in lung cells by 2 times.

In the variants of pre- and post-treatment with an antimutagenic product against the background of cadmium iodide (options 7 and 8), the antimutagenic effect in bone marrow cells is pronounced - the level of chromosomal aberrations is reduced by 4 times, and the level of nuclear abnormalities in the cells of the lungs and bladder is reduced by 3 colon in 2.6 times compared with the positive control (option 2).

Thus, the antimutagenic product showed the highest modifying effect at a dose of 4-6 g / kg (based on 1 kg of the animal’s body weight) not only against the background of positive control in the pre- and post-treatment options, but also against the background of spontaneous mutagenesis

Example 3

The berries of gooseberry ordinary (Ribes uva-crispa Ζ.) And Jerusalem artichoke tubers (Helianthus tuberosus L.) are subjected to preliminary cleaning and washing, then they are chopped and, using compression, separate diffusion juice of berries and vegetables is obtained. Before blending, the juices are concentrated, evaporated to reduce the volume by 2 times, then mixed in a 1: 1 ratio, followed by evaporation for 3-4 hours in a water bath until gelation or solids content of 68-70%. The semi-finished product is poured into molds and cooled at room temperature for 10-12 hours. Thus obtained antimutagenic product in the form of marmalade is freed from molds and Packed in plastic bags.

In this series of experiments, we studied an antimutagenic product based on the juice of Jerusalem artichoke tubers and gooseberry berries of 6-8 g / kg (based on 1 kg of the animal’s body weight), against the background of negative and positive controls, in variants of pre- and post-treatment with a model mutagen - cadmium iodide An analysis of the experimental results presented in Table 1 showed that the level indicators of both chromosomal aberrations and abnormalities of the nucleus of variants 9, 10, 11 are statistically close to the indicators of options 6, 7, 8.

Thus, the use of an antimutagenic product in a dose of 6-8 g / kg (based on 1 kg of the animal’s body weight) is physiologically not feasible and economically disadvantageous.

An analysis of the experiment indicates that

1. The antimutagenic product based on juices from Jerusalem artichoke tubers and gooseberries has a positive modifying effect, reducing the clastogenic effect of cadmium in the cells of the bone marrow, lungs, bladder and colon.

2. Juices from Jerusalem artichoke tubers and gooseberries in a jelly-like state (marmalade) exhibit high antimutagenic activity at a dose of 4-6 g / kg (based on 1 kg of animal body weight).

3. It is advisable and cost-effective to recommend an antimutagenic product in a dose of 4-6 g / kg (based on 1 kg of the animal’s body weight) for recreation of the organism genotype when contaminated with heavy metals using cadmium as an example.

Claims (1)

A method of preparing an antimutagenic product in the form of marmalade from a mixture of common gooseberry juices ( Ribes uva-crispa L. ) and Jerusalem artichoke tubers ( Helianthus tuberosus L. ), which involves the preparation of diffusion juices of common gooseberry and Jerusalem artichoke by pressing, concentrating the juices by evaporation to reduce the volume by 2 times, mixing concentrated juices in a volume ratio of 1: 1 and evaporating for 3-4 hours in a water bath until gelation or solids content of 68-70%.