SU1462196A1 - Method of quantitative analysis of aggregation and disaggregation ability of erythrocytes - Google Patents

Abstract

This invention relates to medicine. The purpose of the invention is to improve the accuracy of the method in the study of plasma with a high content of lipids. Plasma viscosity and plasma viscosity are recorded after the addition of erythrocytes, fibrinogen solution and gammaglrbulin. Then rassgotigoat. erythrocyte aggregation capacity and determine the shear rate characterizing the hydrodynamic disaggregation of erythrocytes. The method allows the study of the rheological properties of blood taking into account the behavior of the cells in the stream.

Description

The invention relates to medicine and can be used to quantify the aggregation and disaggregation of erythrocytes, which are important for the diagnosis of disturbances in the rheological properties of blood in r - d pathological conditions, including coronary heart disease (CHD), as well as for evaluation of the effectiveness of the treatment.

The goal is to improve the accuracy of the method in the study of plasma with a high content of lipids.

The method is carried out as follows.

Blood sampling by puncture of the elbows oh vein in the amount of 9 ml. 9 ml of blood is stabilized by adding 1 ml of 3.8% sodium citrate solution.

Stabilized blood is centrifuged at 1000 rpm for 10 minutes. Then the platelet-rich plasma is separated and centrifuged again at 3000 rpm for 15 minutes. With help . pipettes suck off plasma that does not contain blood cells.

The erythrocyte suspension is prepared in a ratio of 1: 400 in plasma, not containing corpuscles of blood, previously erythrocytes three times in a physiological solution. The amount of erythrocytes in the suspension obtained is monitored on an electronic counter for counting the uniform elements of blood Picoscale PS-4 (the number of erythrocytes) (4300000 + 200000 per 1 ml).

 Preparation of a 5% fibrinogen solution: 90 mg of dry fibrinoreI prepared from human blood is diluted with 2.5 ml of distilled water.

102-u solution of globulin is produced by the Kiev plant of bacterial preparations. The change in plasma viscosity in the amount of 2.7 ml on a micro-: VIR-79 ME electric rheometer in the range of the shear rate of 0.7-121 s.

After measuring the plasma viscosity; 0.1 ml of erythrocyte suspension, 0.2 ml of 5% fibrinogen solution and 0.2 ml of 10% solution, y-globulin is added to it, and the viscosity of the resulting suspension is measured in range of shear rates 0.7-121.0. one

The calculation of indicators is carried out according to the formula

(a-c)

at

AE

100%,

where is AE - aggregative ability

red blood cell percentage; a - - viscosity of erythrocyte suspension in plasma with the addition of fibrinogen and V -glob; :. bulin;

c - plasma viscosity. Determination of shear rate characterizing the hydrodynamic degassing of red blood cells.

The shear rate at which erythrocyte aggregates are destroyed can be determined by calculation or graphical method.

The computational method consists in calculating the percentage increase in viscosity of a suspension of aggregated red blood cells at all fixed shear rates in the range of 0.7-121.0 (i.e., at 0.7, 1.8, 3.8, 6.3; 8; 13; 17; 28; 35; 58; 121). The shear rate at which the difference in magnitude is 6% indicates that the disaggregation has been depleted.

The graphical method consists in constructing a graph of the dependence of viscosity on the shear rate (on the abscissa axis, the values in the body are deposited, on the ordinate axis - on the values of the shear rate). The shear rate at which the difference in viscosity of a plasma lacking uniform elements and the viscosity of a plasma with aggregated

5 red blood cells will be insignificant

(about 6%) indicates disaggregation -. .

Example 1. Patient A. Plasma viscosity at shear rate 0.7

0 2.07 spz.

Plasma viscosity with aggregated erythrocytes at a shear rate of 0, 2.48 spz.

AE

 i2Z2llI222.) x 100%

Thus, the aggregation ability of red blood cells is equal to 20%.

When plotting the dependence of viscosity of pure plasma and plasma viscosity with aggregated erythrocytes on shear rate, it is noted that at 58. With a difference in viscosity, two samples is 6%. Thus, at 58 s, hydro / danamic disaggregation of erythrocytes occurred.

Example 2. Free L.

The plasma viscosity at a shear rate of 0.7 is 1.68 spz.

Plasma viscosity c. aggregated red blood cells at a shear rate of 0.7 s is equal to 1.9 spz.

AE

From here

in (spz) 100%

. T,

 20%.

Erythrocyte aggregation ability is 24%.

When plotting the dependence of the viscosity of the pure plasma and the viscosity of the plasma with the aggregated erythrocytes on the shear rate, it is noted that at 13 the difference in viscosity of two samples is 6%. This indicates that at 13 hydrodynamic disintegration of erythrocytes occurred,

PRI me R 3. Patient H.

The plasma viscosity at a shear rate of 0.7 s is 1.45 sps.

Plasma viscosity with aggregated erythrocytes at a shear rate of 0.7 s is 2, 1 sps.

Uh

From here. (a), - 100%

  - spz) 100% ./ „, 51

Erythrocyte aggregation ability is 44%.

When plotting the sasiiCHMost viscosity of bestrombotditarny plasma and plasma viscosity with aggregated erythrocytes on the shear rate, it is noted that at a shear rate of 58 s, the difference in viscosity of the two media is 6%, i.e. shear rate 58 characterizes the hydrodynamic disaggregation of red blood cells.

The proposed method allows to quantitatively determine the aggregation capacity and disaggregation properties of red blood cells, taking into account the behavior of cells in the stream. The method is simple, not only does not require additional material costs and expensive equipment, but also allows using a rotary meter in a zizimeter with great economic efficiency, expanding the scope of its application in studying the rheological properties of blood.

62196

Formula

the invention

The method of quantitative determination of the aggregation and deaggregation capacity of erythrocytes, including plasma separation and adding erythrocyte suspension to it, processing the resulting mixture with fibrinogen solutions i) r -globulin, I distinguish it by the fact that, the content of lipids, the plasma viscosity is recorded, and the resulting mixture and the aggregation capacity of erythrocytes are determined by the percentage increase in viscosity of the aggregate suspension relative to the quickest viscosity botsitarnoy plasma

at shear rates of 0.7–13 s, and the disaggregation ability of erythrocytes is the magnitude of the shear rate, which causes hydrodynamic disaggregation of cells.

Claims (1)

Claim A method for quantitative determination of the 5 aggregation and deaggregation ability of erythrocytes, including ¹ plasma separation and adding erythrocyte suspension to it, processing the resulting mixture with solutions of fibrinogen-10 per and J-globulin, characterized in that, in order to increase the accuracy of the method when studying plasma with a high content lipids recorded platelet plasma 2 ° at shear rates of 0.7-13 s ~ ', and erythrocyte disaggregation ability - according to the shear rate, which causes hydrodynamic disaggregation of cells.