RU2427325C1 - Method of estimating efficiency of treatment of chronic placental insufficiency in experiment - Google Patents

Abstract

FIELD: medicine. ^ SUBSTANCE: invention relates to field of medicine, namely pathophysiology, obstetrics, perinatalogy, and can be used for estimation of efficiency of treatment of chronic placental insufficiency. For this purpose in experiment in pregnant rats, starting with 10-th day of gestation, daily for 2 hours chronic placental insufficiency is modeled in pressure chamber of flow type in conditions corresponding to height of 6000 metres. Half of the rats are given a subcutaneous injection of medication "Traumel C" every second day in dose 0.2-0.3 ml for 12-13 days. After that, on 23-24 day of pregnancy rats are subjected to euthanasia and morphometric study of fetuses and placentas and histological study of placentas are carried out. If 25 % retarded weight gain in females, 25 % and 17.5 % reduction of respectively fetus and placenta weight with comparison with norm, 2-fold reduction of placenta thickness are determined, development of chronic placental insufficiency is stated. Absence of mentioned above changes at the background of introduction of "Traumel C" medication, testifies to efficiency of performed therapy. ^ EFFECT: method provides possibility to study chronic placental insufficiency under certain conditions of hypoxia modelling, as well as treatment optimisation by using such preparation of natural origin as "Traumel C". ^ 2 tbl

Description

The invention relates to medicine, namely to pathophysiology, obstetrics, perinatology, and can be used to evaluate the effectiveness of the treatment of chronic placental insufficiency.

An adverse course of pregnancy, regardless of etiological factors, most often leads to the development of chronic fetal hypoxia and, as a result, to chronic placental insufficiency (FPF). Given the frequency of perinatal complications, among which chronic FPI and fetal hypoxia, occupy one of the leading places, the development of new methodological approaches for studying pathogenesis and the use of effective pharmacological correctors are of undoubted interest.

The mechanisms of action of hypoxia on body cells have been studied quite well. As a result of hypoxic effects, the normal balance of neurotransmitters (glutamate, dopamine, serotonin, acetylcholine, etc.) and their metabolic products in especially sensitive structures changes or the structural and functional properties of cell membranes are violated, which in itself can lead to cell death. In addition, hypoxia changes the functioning of the genetic apparatus of the cell and can initiate the transcription of specific genes responsible for programmed cell death. Moreover, pathological changes in the central nervous system depend on the duration of the hypoxic effect and on the stage of ontogenetic development of the central nervous system, at which this effect was applied.

Prenatal hypoxia, like maternal stress, leads to impaired formation of behavioral reactions, the development of motor activity, a weakening ability to learn, and a decrease in body weight of offspring. Oxygen deficiency during pregnancy reduces adaptive capacity in the postnatal period, causing the death and birth of physiologically immature and lagging behind offspring with severe and long-term neurological complications.

From the practice of medicine, there are known methods of treating placental insufficiency: the use of Litovit (RU 2228749 C1, 05.20.2004), intravenous administration of nimotol and nimodipine (RU 2270011 C2, 07/10/2005), the use of enterosorbent against the background of drug therapy (alginic acid sodium-calcium) in addition to the traditional treatment (RU 2149592 C1, 05.27.2000), the use of a phytopharmacological preparation (alcohol extract of phytoecdysteroids) according to a 3-stage scheme (RU 2188661 C1, 09/10/2002).

The disadvantages of these methods is the high drug load on the pregnant woman, the presence of allergic reactions and side effects during treatment, there are contraindications for use.

The problem of introducing methods of influence on the mother and the fetus, which reduce the drug load, remains very urgent. Among them, regulatory methods of treatment, such as antihomotoxic therapy (homotoxicology), which are a synthesis of modern medical advances and a homeopathic approach to treating patients, should be noted. This method made it possible to logically and practically justify and implement the principles of holistic medicine, which provides an integrated approach to treating a person as a single biological system. Treatment with antihomotoxic drugs combines the best traditions represented by both homeopathy and allopathic medicine. The high effectiveness of these drugs, safety (the absence of adverse and allergic reactions), the absence of contraindications and age restrictions make it possible to widely use them in pregnant women and newborns.

The basic principles of homeopathy are the law of similarity, the principle of "small doses" and the principle of an individual approach.

Among the most widely used drugs is the complex antihomotoxic drug Traumeel S, which is created on the basis of natural components, with a pronounced anti-inflammatory, antiexudative effect. It improves microcirculation in any organ and system, gently affects the immune system. This drug is well combined with other antihomotoxic drugs, which increases the overall therapeutic effectiveness of the clinical use of the drug "Traumeel C".

The antihomotoxic drug "Traumeel S" is available in the form of tablets, drops, in ampoules for intramuscular and intravenous administration, as well as in the form of an ointment for external use. The constituent components of the preparation: Arnica D2, Calendula D2, Hamamelis D2, Millefolium D3, Atropa belladonna D4, Aconitum D3, Mercurius solubilis Hahnemanni D8, Hepar sulphuris D8, Chamomilla recutita D3, Symphytum D8, Bellis perechtius Dereniforidae , Hypericum D2.

The mechanism of action of the drug Traumeel C is formed from the complex effect of its components and is characterized by:

- anti-inflammatory and antiviral effects;

- increased vascular tone, elimination of venous stasis and antithrombotic effect, stop bleeding, normalization of prothrombin in the blood serum;

- activation of cellular respiration and oxidation processes;

- stimulation of the body's defense systems;

- activation of wound healing and overcoming the effects of shock;

- analgesic effect;

- restoration of neuromuscular conduction.

The aim of the proposed invention is to optimize the treatment of chronic placental insufficiency through the use of drugs of natural origin.

The goal of the invention is achieved in that in pregnant rats, starting from the 10th day of gestation, chronic placental insufficiency in a flow chamber is simulated daily for 2 hours, under conditions corresponding to an altitude of 6000 meters, half of them are injected subcutaneously after 1 day 0, 2-0.3 ml of the complex homeopathic preparation "Traumeel C" for 12-13 days, then on 23-24 days of pregnancy all females are euthanized and a morphometric study of the fetus and placenta, histological examination of the placenta and on the basis of a delayed increase in body weight of females by 25%, a decrease in the weight of fetuses and placenta by 25% and 17.5%, respectively, compared with the norm, a decrease in the thickness of the placenta by 2 times indicate the development of chronic placental insufficiency, and in the absence of the above changes against the use of the drug of natural origin "Traumeel S" indicates the effectiveness of the therapy.

The proposed method has been successfully tested on 40 laboratory outbred white rats, females, 10 animals made up the control group (group 1), 20 animals that underwent hypoxia from 10 to 22 days of gestation were divided into 2 equal groups: animals of the 2nd group treated with the drug "Traumeel C" 0.2-0.3 ml / m after 1 day; 3rd animals - groups that did not receive the Traumeel S preparation. The fourth group included 10 animals that received the drug "Traumeel C", but not exposed to hypoxia. Hypoxia was modeled in a 10-liter flow chamber. The rats of the main observation group were raised to a height of 6000 m at a speed of 50 m / s. Ascent rate was estimated using an airplane altimeter and an altimeter. The exposure at a height of 2 hours. A 40% KOH is used as a CO ₂ absorber.

Before the start of the experiment, the animals were kept in quarantine (1 month) with obligatory clinical examination and culling of suspicious individuals. The experimental protocol in the sections on the selection, keeping of animals, modeling of pathological processes and their derivation from experience was compiled in accordance with the principles of bioethics, laboratory practice rules (GLP); it complies with the ethical standards set forth in the Geneva Convention (1971), “International Recommendations for Biomedical Research Using Laboratory Animals” (1985) and in accordance with the order of the Ministry of Health of the Russian Federation No. 267 of 06/19/2003 “ On the approval of the rules of laboratory practice ”(Appendix to the order of the Ministry of Health of the USSR of 08.08.1977, No. 755).

To avoid premature birth, the females were subjected to ephtanasia by intraperitoneal administration of 40 mg / kg of sodium ethamine at 22-23 days of gestation, a morphomeric study of the fetus and placenta was performed, as well as a histological examination of the placenta. The material was preserved in 10% formalin. After standard histological treatment, the tissue was embedded in paraffin. Preparations made from paraffin blocks were stained with hematoxylin and eosin. The study was carried out by examining a serial section under an electron microscope.

Statistical processing of the results was carried out using Microsoft Exel 7.0 for Windows 98 and Sigma Stat 3.0. The data were analyzed using descriptive statistics that evaluated the arithmetic mean and the arithmetic mean error. The hypothesis of equality of means in the cases with normal distribution was tested using the Student criterion. In other cases, the nonparametric Mann-Whitney-Wilcoxon test was used. Qualitative traits were evaluated according to the Fisher criterion. Differences were considered statistically significant at a 5% significance level.

Indicators of the prenatal period under the influence of hypoxia (M ± m) are presented in table 1.

With a histological examination in the placental tissue, you can detect all the layers that are normal in the rat chorioallantoic placenta (see diagram).

The main layer of placenta in thickness is the labyrinth layer, in which a large number of sharply full-blooded anastomosing vessels were detected.

The next most pronounced placental tissue was the layer of spongiotrophoblast, in which compactly located clusters of basophilic cells of spongiotrophoblast were determined. Spongiotrophoblast cells surrounded by a round and irregular shape fused foci, represented by light cells with a pycnotic nucleus, and containing dilated full-blood vessels with a thin wall.

The decidual layer was the most peripheral layer of placental tissue and was represented by a thin layer of predominantly flattened decidual cells. The layer of the chorionic plate was not always clearly distinguishable. It was represented by edematous fibrous tissue with large full-blooded vessels (see Table 2).

The calculation and discussion of the obtained measurement results of the layers was carried out based on the thickness of the main three layers of the placenta: the labyrinth layer, the layer of spongiotrophoblast and decidual layer, the ratio of the thickness of the three main layers of the rat placenta in%.

Studies conducted on this model showed that against the background of prenatal hypoxia, there was a delayed increase in body weight in females by the 22nd day of pregnancy in the 2nd group by 12.5% (p> 0.05), and in the 3rd - by 25, 2% compared to normal (p <0.05). When registering somatic indicators, it was found that the weight of the fetus was significantly reduced compared with the control in animals of the 3rd group by 25.8% (p <0.05), and the placenta weight in the 3rd group of observation was 17.5% less. than in the control (p <0.05). The data obtained are consistent with literature data indicating that the reduction in placenta mass depends on the severity and duration of the pathological process (see table. No. 1). The increase in placenta mass in the 4th observation group, mainly due to the labyrinth layer (0.53 ± 0.02 versus 0.51 ± 0.05 in the control), is due to the stimulating effect of the Traumeel C preparation (p> 0.05 )

At the same time, a significant decrease in the mass and volume of the placenta is more often observed during pregnancy, complicated by intrauterine growth retardation, and the degree of decrease clearly correlates with the severity of fetal malnutrition. Such changes indicate a violation of the trophic function of the placenta and the development of secondary placental insufficiency. Therefore, of particular importance is the study of the morphological structure of the rat placenta under hypoxia, as well as the effect of drugs of natural origin on the structure and function of the placenta.

The maximum thickness of the placenta was observed in 1 and 4 of the studied groups (2546.91 ± 177.51 and 2787.76 ± 199.79 μm, respectively). A significant decrease in placental thickness was revealed in the 3rd group of observation by 34.1% (p <0.001), in the second group by 17.8% (p> 0.05).

The maximum thickness of the labyrinth layer was observed in groups 1 and 4, while in groups 2 and 3 the thickness of the labyrinth layer was 1.5 and 2 times less (p <0.05) and (p <0.001), respectively. The labyrinth layer is the main and extremely important for supplying the embryo with nutrients and gas exchange.

The layer of spongiotrophoblast to a lesser extent affects the metabolic processes. Its thickness was in the range of 400-600 µm. The minimum values were obtained in group 3 - 442.45 μm (p> 0.05), and the maximum in 4 studied groups - 562.18 μm (p> 0.05).

Decidual tissue thickness ranged from 150 to 300 pm. Its maximum thickness was found in the 3 studied groups - 262.48 µm (p <0.05) compared with the control. In the other three groups, the thickness of the decidual layer was approximately the same (151.72 μm in group 1, 152.34 μm in group 4 and 172.32 μm in group 2).

The measurement results, presented in relative values as the ratio of the layers and expressed in%, confirmed the trends that were observed when comparing the absolute values obtained by measuring the layers in the studied placentas.

The maximum thickness of the chorial plate with the possibility of measurements was 235.76 μm, the minimum 132.51 μm. It was not possible to carry out measurements in all cases.

A similar situation was repeated with respect to the layer of giant cells. This was due to the fact that this layer had no clear boundaries and consisted of 2-3 separately located cells with abundant cytoplasm and a single large nucleus with a well-defined nucleolus.

Therefore, we can conclude that chronic hypoxia in the second half of pregnancy leads to a delay in fetal growth and development of the fetus, a decrease in placental indices, which indicates the development of placental insufficiency.

Thus, the experimental studies conducted on this model showed that prenatal hypoxia has a dramatic negative effect on the course of pregnancy, placenta formation, fetal development, which results in intrauterine growth retardation and fetal development, and a decrease in placental parameters indicating the development of placental insufficiency. At the same time, the use of the complex antihomotoxic drug Traumeel C during pregnancy, which occurs against the background of chronic hypoxia, improves the course of pregnancy, optimizes the physical development of the fetus, stimulates the development of compensatory-adaptive mechanisms in the placenta, leading to an increase in its mass, and has a positive effect on the formation of the placenta. The complex antihomotoxic drug Traumeel S, which contains herbal and natural products used in small doses, which makes this drug completely safe and non-toxic, can be used in pregnant women for the prevention and treatment of chronic placental insufficiency.

Table number 1. Indicator The control 2nd group 3rd group 4th group Weight gain by day 22 of pregnancy (g) 66.3 ± 4.7 58.0 ± 6.3 49.6 ± 4.9 * 64.8 ± 5.8 The mass of fruits (g) 3.1 ± 0.07 2.9 ± 0.09 2.3 ± 0.02 * 3.3 ± 0.1 The mass of the placenta (g) 0.51 ± 0.05 0.48 ± 0.03 0.42 ± 0.01 * 0.53 ± 0.02 Note. Here and in Table 2: * - the data are statistically significant in relation to the control (p <0.05); ** - (p <0.001).

Claims (1)

A method for evaluating the effectiveness of treatment of chronic placental insufficiency in an experiment, characterized in that in pregnant rats, starting from the 10th day of gestation, chronic placental insufficiency in a flow chamber is simulated daily for 2 hours under conditions corresponding to an altitude of 6000 m, half of them subcutaneously through 1 day injected 0.2-0.3 ml of the complex homeopathic preparation "Traumeel C" for 12-13 days, then on 23-24 days of pregnancy all females are euthanized and morphometric examination fetal and placenta, histological examination of the placenta, and on the basis of a delayed increase in female body weight by 25%, a decrease in the weight of the fetus and placenta by 25% and 17.5%, respectively, compared with the norm, a decrease in the placenta thickness by 2 times indicate the development of chronic placental deficiency, and the absence of the above changes on the background of the use of the drug of natural origin "Traumeel C" indicates the effectiveness of the therapy.