RU2249212C1 - Method for predicting gestosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves measuring carbonyl protein derivatives concentration in pregnant woman blood serum at early stage (beginning from 5 weeks). The value being equal to 51.7 optic density units/g of protein or higher, gestosis development is predicted.

EFFECT: high prognosis accuracy in preclinical period.

Description

The invention relates to medicine, namely to obstetrics, and will find wide application for objective prediction of the development of gestosis.

Despite the undoubted success in the treatment of gestosis, it still remains one of the most common complications of pregnancy. The pathochemical and pathophysiological mechanisms of gestosis are already established in the initial stages of pregnancy and are caused by a violation of trophoblast invasion and gestational transformation of the spiral uterine arteries with subsequent damage to the vascular endothelial function. The main problem when deciding on the presence of gestosis is the lack of criteria by which its development is predicted in the very early stages of pregnancy.

An analysis of the patent and medical literature has revealed that for the diagnosis of the most serious pregnancy complication - gestosis, tests related to physiological, morphological, immunological and biochemical methods of research are currently proposed.

1. Physiological tests.

So in the work of A.N. Strizhakova, O.R.Baeva, I.V. Ignatko (Prediction of the development of gestosis and placental insufficiency. Russian Bulletin of the obstetrician-gynecologist. T.1. No. 1. 2001, p. 39-43) is emphasized the possibility of predicting gestosis with Doppler studies in 20-21 weeks., When there is an increase in Doppler indices of vascular resistance in the uterine and umbilical arteries. In the spiral arteries, these changes were noted at 16-19 weeks. A known method for the diagnosis of late toxicosis of pregnancy (copyright certificate No. 1217349, 1986), according to which the differentiation of the severity of toxicosis is determined by the degree of filling of the pulmonary circulation by measuring the amplitude of the systolic wave reopulmonogram in six areas of the lungs. There is a method of preclinical stage of gestosis with chronic prielonephritis (application No. 98111316, 2000), in which the method of ultrasound dopplerometry based on the increase in C / D ratio in the arteries of the kidney parenchyma in 17-21 weeks diagnose the preclinical stage of gestosis.

2. Morphological tests.

Known RF patent No. 2138805, 1999 - a method for the diagnosis of gestosis by evaluating the red blood cell adrenoreactivity of pregnant women.

Known RF patent No. 2143694, 1999 - a method for diagnosing gestosis by determining HCT-positive forms and neutrophil activation index.

3. Immunological tests.

Known methods for the diagnosis of late gestosis:

- according to the author's certificate of the USSR No. 1398832, 1988, the essence of which is to determine in the blood the number of lymphocyte placenta sensitized to antigens and the level of cortisol in a period of 20 weeks;

- according to the author's certificate of the USSR No. 1683672, 1991, according to which the determination of the titer of homoreactants and the titer of natural antibodies to sheep erythrocytes from 24 weeks of pregnancy allows predicting the development of gestosis;

- according to the patent of the Russian Federation No. 2120636, 1998 makes it possible in II-III trimesters by determining immunocompetent proteins in the mother's blood serum to diagnose late gestosis;

- according to the patent of the Russian Federation No. 2135999, 1999 by definition in the blood serum in the period of 13-14 weeks of antiphospholipid antibodies.

4. Biochemical tests.

Known methods for the diagnosis of late gestosis of pregnancy:

- according to the author's certificate of the USSR No. 572534, 1990 to determine the levels of lipids and phospholipids in critical periods for pregnancy, the rates of which increase with pregnancy toxicosis;

- according to USSR author's certificate No. 1358936, 1987, which indicates that toxicosis is diagnosed by protein-free plasma photometry at a wavelength of 254 nm and an increase in optical density by more than 13%;

- according to the author’s certificate CCCP No. 1337778, 1987, according to which the time of erythrocyte hemolysis, the percentage of cell death and the level of free plasma hemoglobin in the peripheral blood are determined to diagnose gestosis and its various degrees;

- according to the patent of the Russian Federation No. 1612263, 1990, which determines the activity of superoxide dismutase (SOD) in the blood serum of a pregnant woman and diagnoses the presence of gestosis;

- in the patent of the Russian Federation No. 2133037, 1999, the determination in plasma of the activity of cathepsin G is proposed, which makes it possible to diagnose gestosis;

- according to the patent of the Russian Federation No. 2138817, 1999, the definition of salivary-sperm globulin in the saliva of pregnant women in the period from 28 weeks, allows you to diagnose late gestosis;

- according to the patent of the Russian Federation No. 2190848, 2002, which indicates a periodic blood test by spectral fluorescence analysis of the extract of coproporphyrins and protoporphyrins of erythrocytes and with a ratio of less than 10, gestosis is diagnosed;

- according to the patent of the Russian Federation No. 2021607, 1994. In the present invention, the diagnosis of late gestosis is based on the calculation of an intoxication index, the composite indicators of which are: the urea content in the blood serum, the concentration of medium-weight molecules and the platelet content. Diagnostic periods are possible from 27 weeks of pregnancy;

- according to the patent of the Russian Federation No. 2040794, 1995 - determination of the resistance of red blood cells to a number of hemolytics. Terms of a possible diagnosis from 23 weeks of pregnancy.

The main disadvantage of the above methods is the impossibility of preclinical diagnosis of gestosis.

The “Method for the diagnosis of gestosis, described in the article“ A modern view of the concept of the development of multiple organ failure in gestosis (I.V. Medvedinsky, I.D. Yurchenko, L.N. Pestryaeva et al. // Collection of scientific works. Perinatal anesthesiology and intensive care of mother, fetus and newborn. Ekaterinburg. - 1999. P.25-32).

The essence of the prototype method is that the determination in the blood of an elevated level of endotheliocytes, indicating damage to the vascular endothelium, confirms the presence of gestosis.

This method, like the previous ones, has a number of imperfections:

1. Impossible early preclinical prognosis of gestosis.

2. The information content of the method is quite high only with increasing severity of gestosis.

These disadvantages are eliminated in the claimed invention.

The objective of the invention: the development of a method that allows with high accuracy in the early stages (at the preclinical stage) of pregnancy to predict the development of gestosis.

The problem is solved in that in the early stages (from 5 weeks) in the blood plasma of a pregnant woman determine the level of carbonyl derivatives of proteins and with a value equal to 51.7 units / g and above, the development of gestosis is predicted.

According to the most common theory, gestosis forms very early, even during placentation, when generalized damage to the endothelium of the decidual membrane and chorionic villi occurs. One of the important causes of damage to the endothelium is a modification of its protein component, in particular, oxidative degradation of proteins, leading to an increase in their carbonyl derivatives. The result of this destruction, in the end, is alteration of the endothelium, accompanied by a violation of vascular permeability and a change in their vasoactivity.

Example 1. C-ba, 26 years old. Case history No. 31. It was examined on an outpatient basis on January 11, 2001. The first pregnant. Dz: pregnancy 5 weeks., Suspicion of an ectopic pregnancy.

The content of chorionic gonadotropin (CH) in the blood serum was determined for the diagnosis of ectopic pregnancy (negative test). The degree of oxidative degradation of proteins in blood plasma was also determined by the level of carbonyl derivatives on a PV 125 1C Solar spectrophotometer, Belarus (Dubinina E.E., Burmistrov S.O., Khodov D.A., Porotov I.S. Oxidative modification of serum proteins human blood.Methods of its determination // Issues of Medical Chemistry. - 1995. - T.41, No. 1. - P.24-26).

0.75 ml of 1/15 M phosphate buffer (pH 7.4), 0.1 ml of a mixture of 4 mM FeSO ₄ solution with 1 mM EDTA solution (1: 1) are added to 0.05 ml of plasma (control and experimental samples) and 0.1 ml of a 0.3 mm solution of H ₂ O ₂ . After 15 min of incubation at a temperature of 37 ° C in a water bath, 1.0 ml of a 0.01 M solution of 2,4-DNPH (2,4-dinitrophenylhydrazine) dissolved in a 2 M Hcl solution are poured into the experimental samples. Instead of 2,4-DNPH, an equal volume of a 2 M HCl solution is added to the control samples. To precipitate proteins, 1 ml of a 20% TCA solution is added to each sample. Samples are incubated at room temperature for one hour and then centrifuged at 3000 g for 15 minutes. Next, the precipitate is washed 2 times with a mixture of ethyl alcohol and ethyl acetate in a ratio of 1: 1. The resulting precipitate was dried in air and then dissolved in 3 ml of an 8 M urea solution. The optical density is recorded at a wavelength of 363 nm. The level of oxidative modification of proteins is expressed in units of optical density per 1 g of protein.

The level of oxidative degradation was 51.7 u / g protein. This made it possible to predict the development of late pregnancy toxicosis in a pregnant woman.

06/18/2001, a pregnant woman was admitted to the department of pathology of pregnant women. D-z: pregnancy 21 weeks., Nephropathy II degree (gestosis EN).

This clinical example convincingly shows that an increase in oxidative degradation of proteins above the boundary indicates the possibility of predicting gestosis.

Example 2. I-ko, 24 years old. Case history No. 33. It was observed in the clinic RNIIAP. Dz: pregnancy 5-6 weeks. On January 6, 2000, blood was taken from a vein, in the plasma of which the oxidative degradation of proteins was determined by the content of carbonyl groups. The level of oxidative modification of proteins corresponded to 51.6 units / g of protein, confirming its normal value. Such indicators of oxidative destruction of plasma proteins indicate the normal formation of chorion.

Further monitoring of the woman made it possible to establish that her pregnancy and childbirth proceeded without complications.

This clinical example indicates the absence of gestosis with oxidative destruction values below the boundary.

According to the claimed method examined 341 women, among whom 23 in the period of 4-5 weeks. In pregnancy, the degree of oxidative degradation of proteins ranged from 51.7 units / g protein to 83.0 units / g protein. In 318, the value of oxidative degradation of proteins was from 45.1 units / g of protein to 51.6 units / g of protein. All pregnant women with protein modification rates of 51.6 units / g protein or lower did not develop gestosis. In women with an oxidative degradation level of 51.7 units / g of protein and higher, late gestosis occurred.

The above confirms the prognostic value of the proposed method.

Thus, the claimed method is more efficient compared to the known and has several advantages:

1. The method is highly specific (determining the level of destruction of the protein component of the cell membrane of the vascular endothelium, including the vessels of the decidual membrane and chorionic villi, reflects the degree of violation of trophoblast invasion, which underlies the development of gestosis).

2. It has high accuracy. False negative and false positive indicators - 0.

3. Detection of pathology in the early stages of pregnancy, starting from 5 weeks of gestation.

4. Clinical application of the method will allow timely implementation of adequate obstetric tactics.

Claims (1)

A method for predicting preeclampsia by examining a pregnant woman’s blood plasma, characterized in that in the early stages (from 5 weeks of gestation) the level of carbonyl derivatives of proteins is determined and the development of preeclampsia is predicted at a value of 51.7 units of optical density / g protein.