RU2309412C1 - Method for evaluating the degree of gestosis severity - Google Patents

Abstract

FIELD: medicine, obstetrics.

SUBSTANCE: starting from the 5^th wk of pregnancy it is necessary to detect the content of endothelin-1 (ET) and sodium oxide metabolites (nitrites and nitrates - NOx) in blood serum. At NOx content ranged 20.9-24.8 mcM/l, and ET-1 ranged 0.7-3.9 pkg/ml one should conclude upon the development of light-degree gestosis, and at NOx value ranged 24.8-49.6 mcM/l, and ET-1 ranged 4.0-8.3 pkg/ml one should diagnose average-degree gestosis. The innovation suggested enables to decrease pharmacological impact upon a mother and a fetus due to prescribing up-to-date adequate therapy of gestosis that corresponds to its severity degree.

EFFECT: higher accuracy of evaluation.

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Description

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

To date, there is a problem in predicting the severity of gestosis, which is one of the serious complications of pregnancy, which takes a leading place among the causes of maternal and perinatal mortality (S.D. Motsar, V.V. Malyuk. Status of the homeostasis system in pregnant women with gestosis on the background the use of fraxiparin (http://www.placenta.mk.ua/spek.next.html).

The treatment of preeclampsia is aimed at eliminating generalized vasospasm, lowering blood pressure, normalizing general hemodynamics and microcirculation in vital organs, correcting metabolic disorders and restoring homeostasis, thereby achieving the necessary level of fetal vital activity.

Timely diagnosis of the severity of disorders in preeclampsia (the severity of this complication of pregnancy) underlies the choice of therapy necessary for this type of preeclampsia, which allows you to adequately influence the clinical manifestations without harming the fetus.

An analysis of the patent and scientific and medical literature has allowed us to establish that tests related to physiological, morphological, immunological and biochemical methods of research are currently proposed for the diagnosis of gestosis.

The work of A.N. Strizhakova, O.R.Baeva, I.V. Ignatko (Predicting the development of gestosis and placental insufficiency. Russian Bulletin of the obstetrician-gynecologist. T.1. N1. 2001. S.39-43) emphasizes the possibility of predicting preeclampsia with Doppler studies at 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.

The following analogues can be attributed to biochemical:

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

- USSR author's certificate No. 1337778, 1987, according to which the peripheral blood determines the time of erythrocyte hemolysis, the percentage of cell death and the level of free plasma hemoglobin, allowing to diagnose gestosis and its various degrees,

- USSR author's certificate No. 1572534, 1990 for determining lipid and phospholipid levels in pregnancy-critical periods, the rates of which increase with pregnancy toxicosis,

- RF patent No. 1612263, 1990 - determine the activity of superoxide dismutase (SOD) in the blood serum of a pregnant woman and diagnose the presence of gestosis,

- RF patent No. 2021607, 1994 - 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,

- RF patent No. 2040794, 1995 - determination of the resistance of red blood cells to a number of hemolytics; terms of possible diagnosis from 23 weeks of pregnancy,

- RF patent No. 2133037, 1999 proposes the determination of plasma activity of cathepsin G, which makes it possible to diagnose gestosis,

- RF patent No. 2138817, 1999 - the determination of salivary-sperm globulin in the saliva of pregnant women in the period from 28 weeks allows you to diagnose late gestosis,

- RF patent No. 2190848, 2002, which indicates a periodic blood test by spectral-fluorescence analysis of the extract of coproporphyrins and protoporphyrins of red blood cells, and with a ratio of less than 10, gestosis is diagnosed,

- there is evidence of a decrease in the amount of α-fetoprotein in the blood serum of a pregnant woman adequate to the severity of gestosis (V.G. Levchenko et al. Alpha-2-fetoprotein in uncomplicated pregnancy and gestosis: www.soramn.ru/Journal/2004/№4),

- RF patent No. 2263913, 2005 - to determine the activity of cathepsin D in venous blood from 6-13 weeks and with its increase, the development of gestosis in the third trimester is predicted.

These methods for the diagnosis of gestosis have significant disadvantages:

1. The diagnosis of gestosis starting only from 16-19 weeks of pregnancy, without taking into account the severity.

2. The use of biochemical markers reflecting metabolic changes resulting from gestosis.

3. Evaluation of membrane disorders available for cell research, a change which can occur not only with gestosis, but also with a deficiency of glucose-6-phosphate dehydroginase.

4. In a number of methods there is a low accuracy of the method.

5. The use of lysosomal enzymes that reflect the activity of free radical processes for predicting gestosis is not justified, because an increase in their activity is not specific for gestosis and occurs in various diseases.

The prototype of this invention is selected "Method for predicting gestosis" (RF patent No. 2292212, 2005).

The essence of the prototype method is that in the plasma of a pregnant woman in the early stages (from 5 weeks of gestation) determine the level of carbonyl derivatives of proteins and at a value equal to 51.7 units optical density per 1 g of protein (51.7 units of opt. pl. per g / protein), the development of gestosis is predicted.

This method, like the previous ones, has the same number of imperfections.

These disadvantages are eliminated in the claimed method.

The objective of the invention: the development of a method that allows in the early stages of pregnancy to assess the degree of gestosis in order to prescribe a differentiated treatment.

The problem is solved by the fact that from 5 weeks of pregnancy women determine serum biochemical compounds endothelin-1 (ET-1) and nitric oxide metabolites (nitrites and nitrates), with their values denoted by NOx from 20.9 to 24.8 μmol / l, and ET-1 from 0.7 to 3.9 pg / ml evaluate the development of mild gestosis, and with NOx from 24.8 to 49.6 μmol / l, and ET-1 from 4.0 to 8 , 3 PCG / ml diagnosed with moderate gestosis.

The technical result achieved by the invention is that the accuracy of the method is increased, its operability is improved, the method is simplified, the pharmacological effect on the mother and the fetus is reduced due to the appointment of a timely, appropriate severity of gestosis therapy.

The leading link in the pathogenesis of gestosis is endothelial dysfunction. The latter is characterized by a violation of the production of factors that ensure the physiological state of blood vessels. These include the most important vasodilator - nitric oxide (NO) and a vasoconstrictor - endothelin-1 (ET-1).

Markers of endothelial dysfunction can be vasoactive compounds that regulate vascular tone and hemostasis. Such compounds include endothelin-1 and nitric oxide, which have the opposite effect on these processes: endothelin, as is known, has a pronounced vasoconstrictor effect, and nitric oxide has a vasodilator effect (A.F. Vanin, 1998; M. - K. Stokle et al. ., 1998). Inadequate formation of vasoactive substances, reflecting the degree of metabolic modification in the endothelium, is an objective indicator of the severity of the possible development of gestosis, since a violation of the physiological ratio of nitric oxide and endothelin leads to a decrease in the blood flow velocity in the uterine arteries, which may well be an initiating factor in the development of gestosis. The performance of the proposed method is confirmed by the following clinical examples:

Example 1. Pregnant St.-va, 26 years old, medical history No. 31 dated 2.08.05

It was examined on an outpatient basis. The first pregnant. Dz: pregnancy 5 weeks.

The patient underwent ultrasound, the content of chorionic gonadotropin in the blood serum was determined, which confirmed the presence of uterine pregnancy. Nitric oxide (NO) was also determined by the number of its metabolites (NO ₂ ^- and NO ₃ ^- ) using the Griss reaction. To do this, 0.8 ml of blood serum is poured 0.8 ml of a 10% solution of ZnSO ₄ and 0.8 ml of a 0.5 N NaOH solution. Then centrifuged for 10 minutes in a refrigerator centrifuge at 3000 rpm (t + 6 ° C). We select the centrifuge and add the commercial Griss reagent 1: 1 to it. After 10 minutes, the optical density of this sample was measured at 530 nm in a cuvette with a 1 cm layer thickness on a spectrophotometer (SF-530, Russia) against the control, which was a sample treated in a similar way, but containing distilled water instead of serum. The NOx content is calculated from a pre-built calibration schedule. The results are expressed in μmoles of NOx per liter of serum. The NOx level was 24.8 μmol / L, and the endothelin-1 level was 8.3 pg / ml. Our data indicate that this pregnant woman developed moderate gestosis. In physiological pregnancy, the level of NOx varies from 17.5 to 20.8 μmol / L; endothelin - from 0.4 to 0.69 pkg / ml.

Determination of the content of endothelin (ET-1) in blood serum was carried out by enzyme immunoassay using kits from DRG International, Inc. (USA). The results obtained allow us to conclude that a pregnant woman has moderate gestosis.

It was decided to start generally accepted therapeutic measures aimed at eliminating preeclampsia, for which a woman was hospitalized in the Department of Pregnancy Pathology of FGU RNIIAP.

Inpatient treatment lasted 21 days, after which on August 23, 05, a woman's blood serum was determined for the metabolites of NO (NO ₂ ^- and NO ₃ ^- ), as well as endothelin. The value of NOx reached 20.8 mmol / L, and the concentration of endothelin was equal to 0.4 pcg / ml, which indicated that these indicators corresponded to the figures characteristic of physiological pregnancy.

09/30/05, the woman was admitted to an outpatient appointment. Diagnosis: Pregnancy 14 weeks.

The determination of nitric oxide (NO ₂ ^- and NO ₂ ^- ) and endothelin in the blood serum showed that the level of NO is 17.5 μmol / L, and that of endothelin is 0.69 pcg / ml, which indicates the absence of gestosis.

This clinical example shows the efficiency of the method for assessing the severity of gestosis. Regular monitoring of the condition of the woman during the entire gestation period contributed to the normal course of pregnancy and the birth of a healthy baby.

Example 2. Patient M-kova. Case history No. 1002. July 4, 2005 was examined on an outpatient basis. Diagnosis: Pregnancy 5 weeks and 3 days.

Along with general clinical examination methods, the content of NOx and ET-1 in the blood serum was determined. The level of NOx corresponded to 20.9 μmol / L, and endothelin - 3.9 pg / ml.

The obtained determination data for a woman in serum NOx and ET-1 suggest that a mild gestosis is observed in this pregnant woman.

07/19/05, a re-examination. Serum NOx and ET-1 levels reached 24.8 μmol / L and 0.7 pcg / ml, respectively, indicating mild gestosis. These indicators were confirmed by clinical signs: vomiting, drowsiness, periodically appearing headache.

A decision was made to conduct therapy corresponding to a mild degree of gestosis, which makes it possible to minimize the medicinal effect on the fetus. This clinical example shows the efficiency of the proposed method for assessing the severity of gestosis and differentiated therapy. Delivery occurred on time, spontaneously and ended with the birth of a healthy child.

Example 3. Patient Mr. Yang, 22 years old. Case history No. 846. 12.01.05, the Diagnosis: pregnancy of 5 weeks.

After a routine examination, serum NOx and endothelin were additionally determined. The value of NOx was equal to 17.5 μmol / L, and endothelin - 0.4 PCG / ml.

The results obtained indicate a lack of predisposition to the development of gestosis.

Subsequent monitoring of the woman before the onset of labor, which occurred on 10/30/05, confirmed the absence of gestosis.

Thus, the clinical observations presented indicate that the determination of NOx and endothelin in the blood serum allows predicting the development of gestosis of both mild and moderate severity.

According to the claimed method examined 63 pregnant women; 14 of them had mild gestosis, 26 had moderate severity, and 23 had no complications.

In the examined contingent of pregnant women, the content of NOx in the blood serum was from 24.9 μmol / L to 49.6 μmol / L, and endothelin from 4.0 pg / ml to 8.3 pg / ml was found in 26 patients (group 1), who developed gestosis of moderate severity. In 14 examined patients (group 2), the content of NOx in the blood serum was in the range of 20.9-24.8 μmol / L with an endothelin level in the range of 0.7-3.9 pkg / ml. All these pregnant women developed mild gestosis.

23 patients with physiological pregnancy were characterized by lower levels of NOx and endothelin in blood serum and varied in the range: NOx - 17.5-20.8 μmol / L, and endothelin - 0.4-0.69 pkg / ml ( 3 group).

The results of the study, shown in the table, showed that there is a statistically significant difference in the level of NOx and ET-1 in blood serum between the three groups of pregnant women examined. Moreover, these indicators are in non-overlapping diagnostic intervals. The accuracy of the method is 98.7%.

The content of metabolites of nitric oxide and endothelin-1 in blood serum

Indicators Group 1 (moderate severity of gestosis) Group 2 (mild art. Severity of gestosis) 3 group (physiologist. Take) NOx (μmol / L) 24.9-49.6 20.9-24.8 17.5-20.8 ET-1 (pkg / ml) 4.0-8.3 0.7-3.9 0.4-0.69

Thus, the claimed method is compared with the known more informative and has several advantages:

- provides an assessment of the varying degree (severity) of complications of gestosis with high accuracy;

- specific;

- provides an objective differentiated approach to therapy;

- simple to execute;

- has no contraindications.

The method has been tested on a significant volume of material and can be widely used in departments of pathology of pregnant women.

Claims (1)

A method for assessing the severity of preeclampsia by examining a pregnant woman’s blood from 5 weeks of pregnancy, characterized in that serum biochemical compounds of endothelium-1 (ET-1) and nitric oxide metabolites (nitrites and nitrates — NOx) are determined and with a NOx value of 20, 9 to 24.8 μmol / L, and ET-1 from 0.7 to 3.9 pg / ml evaluate the development of mild gestosis, and with NOx from 24.8 to 49.6 μmol / L, and ET-1 from 4.0 to 8.3 pcg / ml diagnosed with moderate gestosis.