RU2630576C1 - Means with hypoglycemic, hypolipidemic and energy protective effects - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: drug with antidiabetic properties on the alloxan diabetes model, which is a dry extract from plant material containing grass of bur beggar-ticks; waterwort grass; flowers of pot marigold; rhizome of elfwort; shoots of bushy marsh cinquefoil; grass of St. John's wort; shoots of blueberry.

EFFECT: drug has effective antidiabetic properties on the model of alloxan diabetes.

4 tbl

Description

The invention relates to the field of pharmacy and can be used as a medicine in the complex treatment and prevention of diabetes mellitus based on a known drug.

As part of the implementation of the state program on import substitution and expanding the list of domestic medicines obtained from plant materials, for the treatment and prevention of diabetes mellitus, the use of the created product based on the extract of the dry seven-component composition of medicinal plants with antioxidant and pancreoprotective activity was found and technically justified (patent No. 2410113 "A drug with antioxidant and pancreatic protection"). The initial plant composition includes: a three-part series (grass); marshmallow cinnamon (grass); Calendula officinalis (flowers); elecampane high (rhizomes); five-leaved shrubbery (shoots); St. John's wort perforated (grass); common blueberries (shoots).

The hypoglycemic, hypolipidemic, and energy protective properties of the claimed drug were investigated in a model of diabetes mellitus caused by a single intraperitoneal injection of alloxan monohydrate into white rats.

The studies were performed on 120 sexually mature white Wistar male rats with an initial weight of 180-200 g. The animals were kept in accordance with the Laboratory Practice Rules (GLP) and the Ministry of Health of the Russian Federation Order No. 708Н dated 08.23.2010 “On Approval of Laboratory Practice Rules”. The experimental work was carried out in accordance with the "Rules for the use of experimental animals" (Appendix to the order of the Ministry of Health of the USSR No. 755 of 08/12/07), "the Rules adopted in the European Convention for the Protection of Vertebrate Animals (Strasbourg, 1986). The research protocol was agreed with the ethics committee of the IOEB SB RAS (protocol No. 3 of February 3, 2015).

Alloxan diabetes in white rats was caused by a single intraperitoneal injection of alloxan monohydrate (Sigma - Aldrich, USA) at a dose of 200 mg / kg against a background of 24-hour food deprivation [6, 7]. One day after alloxan injection, blood glucose levels were determined in all animals using a One Touch Select meter (Johnson & Johnson, USA). Animals whose blood glucose level exceeded 10 mmol / L were divided into 3 groups taking into account age, weight and the principle of randomization.

After determining the blood sugar level, the animals of the first experimental group were intragastrically administered a complex herbal remedy at a dose of 300 mg / kg and subsequently 1 time per day for 20 days. As a reference drug, we used the official phytosport “Arfazetin” [3], prepared in the form of a decoction according to the General Pharmacopoeia Monograph 1.4.1.0018.15 “Infusions and decoctions” of the GF of the XIII edition (2015), which was administered to animals of the second experimental group in a volume of 10 ml / kg per the above pattern. Animals of the control group received purified water in an equivalent volume according to a similar scheme. Studies were performed on days 7, 14, and 21 from the start of alloxan monohydrate administration.

The state of carbohydrate metabolism was assessed by the level of glucose and the content of glycated hemoglobin (Hb _Alc ) in the blood [2]. To assess the lipid-lowering effect of the studied drugs in the blood serum of animals, cholesterol (cholesterol), triacylglycerides (TG), low density lipoproteins (LDL), high density lipoproteins (HDL) were determined by enzymatic photometric method [5]. The study of blood biochemical parameters was carried out on a Sapphire-400 analyzer (Japan) using the Human diagnostic kits (Germany).

The energy state of liver tissue in alloxan diabetes was evaluated by the content of adenosine triphosphate (ATP), lactic (MK) and pyruvic acid (PVA) acids [4].

In the statistical processing of the obtained data, the arithmetic mean (M), the arithmetic mean error (m), the Student criterion (t) and the significance of the differences (P) were calculated. Calculations were performed using the Biostatistics software package. Differences between the compared data were considered significant at a probability of 95% (P≤0.05).

The research data presented in table 1 indicate that the injection of alloxan monohydrate into animals causes the development of severe hyperglycemia. Thus, the blood glucose level in control animals on the 7th day of the experiment increased 4.9 times in relation to the rate of intact animals and remained raised 2.4 and 1.9 times on the 14th and 21st days of observation, respectively.

The studied agent had a pronounced hypoglycemic effect: the blood glucose in animals of the first experimental group during all periods of observation was on average 26% lower than in the control animals. In animals treated with Arfazetin, the blood glucose level on the 7th, 14th and 21st days of observation was 21%, 20% and 24%, respectively, lower than that in animals of the control group.

Against the background of allox diabetes, there was an increase in the content of Hb _Alc in the blood of animals of the control group (table 2). The highest concentration of Hb _Alc , exceeding that in intact animals by 1.7 times, was observed on the 14th day of the experiment.

The course introduction of the studied drugs had a normalizing effect on the carbohydrate metabolism of white rats with alloxan diabetes. So, in animals treated with the claimed drug and Arfazetin, the concentration of Hb _Alc in the blood was lower than that of the animals of the control group on day 7 - by 22% and 20%, on day 14 - by 32% and 29% and on day 21 of the experiment - by 22% and 20%, respectively.

As follows from the data presented in table 3, a single administration to animals of alloxan monohydrate caused a violation of lipid metabolism. So, in animals of the control group on the 7th day of the experiment, the level of cholesterol in the blood serum was 1.6 times higher than that in animals of the intact group, LDL - 2.2 times, TG - 3.9 times, while the content of HDL was 1.4 times lower. Lipid metabolism in animals of the control group returned to normal only by 21 days of the experiment, while the content of TG in the blood serum remained increased by 17% compared with the data of animals in the intact group.

The introduction of the declared herbal remedy into the animals contributed to an earlier normalization of their lipid metabolism in comparison with the data in the control group of animals (table 3). So, on the 7th day of the experiment in animals of the 1st and 2nd experimental groups, the cholesterol content was lower than that in the control animals by 24% and 22%, respectively, the concentration of TG was on average 41%, and the LDL content was 27% and 18%, respectively. Against the background of the introduction of the studied drugs, the HDL concentration was higher by 25% and 20%, respectively, of the data in control animals. In animals of the experimental groups, on the 14th day of observation, a number of indicators of the lipid spectrum (cholesterol, LDL, HDL) almost corresponded to that in intact animals, and on the 21st day normalization of the level of TG in the blood was noted.

Due to significant changes in the metabolism of both carbohydrates and fats, alloxan diabetes causes a violation in the provision of cells with the main substrates of plastic and energy metabolism. From the data presented in table 4, it can be seen that this pathology at all periods of observation is accompanied by a significant decrease in the energy supply of liver tissue. So, on the 7th day of the experiment, a five-fold decrease in ATP content was observed in the control group of animals compared to intact. The content of MK and PVC in the liver homogenate of control animals increased by 2.8 and 1.8 times, respectively, relative to those in intact animals. The increase in the concentration of PVA is probably due to the inhibition of the pyruvate dehydrogenase complex, the activity of which, as is known, decreases with hyperglycemia [8]. As a result, the MK / PVK ratio in the control group was 19/1, against 12/1 in the intact group, which indicates the intensification of anaerobic breakdown of glucose and increased gluconeogenesis. On the 14th day of the experiment, in animals of the control group, energy deficiency in the liver tissue persisted, and its severity was comparable to that on the 7th day of observations. When assessing energy metabolism on the 21st day of the experiment, attention was drawn to an increase in the content of intracellular ATP, as well as a decrease in the content of lactate and pyruvate in the liver tissue of control animals with respect to the data on the 7th and 14th days of observation. However, this increase in the energy status of the organ did not reach the values in animals in the intact group. Moreover, the MK / PVC ratio in the control by 21 days of the experiment was 16/1, which indicates the ongoing suppression of aerobic and strengthening of the anaerobic mechanism of energy synthesis.

In the experiment, it was shown that the claimed agent in the course of administration to animals of the experimental group exerts an energy protective effect in case of alloxan diabetes. As follows from the data presented in table 4, on the 7th day of the experiment, the ATP content increased by 31%, and the concentration of MK decreased 1.7 times compared to the control. In this case, the MK / PVC ratio with the introduction of the new agent was 12/1 against 19/1 in the control group of animals. At a later date of the experiment (14 and 21 days), against the background of the administration of the claimed drug in animals, a significant increase in ATP concentration by 2.3 and 1.2 times, respectively, and a decrease in MK content by an average of 1.5 times compared with the control animals . The MK / PVK ratio reached the physiological norm by the end of the experiment and was 11/1. The indicated energy protective effect is apparently due to a partial restoration of oxidative phosphorylation as a result of a decrease in the production of reactive oxygen species by the liver mitochondria under the action of the claimed agent.

The effect of the comparison drug "Arfazetina" on the energy status of the liver in alloxan diabetes did not exceed the effect of the claimed agent.

Thus, the dry extract from plant materials containing a three-part series (grass); marshmallow cinnamon (grass); Calendula officinalis (flowers); elecampane high (rhizome); five-leaved shrubbery (shoots); St. John's wort perforated (grass); common blueberries (shoots), has a pronounced hypoglycemic, lipid-lowering and energy protective effect in alloxan diabetes. The claimed drug reduces glucose and glycated hemoglobin in the blood, reduces the severity of dyslipidemic disorders, reducing the content of cholesterol, triglycerides and LDL and increasing the level of HDL in the blood serum, as well as stimulating the production of intracellular ATP with a decrease in the severity of acidosis, thereby eliminating the cause of development diabetes mellitus.

Literature

1. The State Pharmacopoeia of the Russian Federation of the XIII edition (2015). http://193.232.7.107/feml.

2. Korolev, V.A. A new method for determining glycated hemoglobin / V.A. Korolev // Journal of Grodno State Medical University. - 2011. - No. 1. - S. 36-41.

3. Mashkovsky, M.D. Medicines / M.D. Mashkovsky. - M: New Wave, 2012 .-- S. 1216.

4. Methods of biochemical studies: lipid and energy metabolism / ed. M.I. Prokhorova. - L., 1982. - 272 p.

5. Methods of clinical laboratory research / ed. B.C. Kamyshnikov. - M .: MEDpress-inform, 2013 .-- 736 p.

6. Guidelines for preclinical studies of drugs. Part one. - M .: Grif and K, 2012 .-- 944 p.

7. Experimental diabetes mellitus / Ed. - V.G. Baranova. - L .: Nauka, 1983 .-- 240 p.

8. Reddi, AS Selenium-deficient diet induces renal oxidative stress and injury via TGF-β ₁ in normal and diabetic rats / AS Reddi, JS Bollineni // Kidney International. - 2001. - Vol. 59, Issue 4. - pp. 1342-1353.

Claims (1)

A drug with antidiabetic properties on the model of alloxan diabetes, which is a dry extract from plant materials containing grass of a tripartite series; marsh grass sushitsy; calendula officinalis flowers; Elecampane rhizome high; shoots of a five-leaved shrub; Hypericum perforatum grass; shoots of common blueberries.