SU1467516A1 - Method of analyzing disturbance of lipoprotein spectrum of blood plasma - Google Patents

Abstract

The invention relates to the field of medicine, namely, clinical and biochemical laboratory diagnostics and cardiology. The aim of the invention is to improve the accuracy of the method by increasing its sensitivity. About 2 ml of blood is taken from the patient on an empty stomach, plasma is separated by centrifugation. The total cholesterol and cholesterol content of α-lipoproteins is determined in plasma. In addition, electrophoretic separation of lipoproteins by polyacrylamide gel electrophoresis is carried out, followed by recording of a densitogram and determining the content of pre-α-lipoproteins, / b-lipoproteins, slowly and rapidly migrating subfractions of α-lipoproteins. After that, the atherogenic index is calculated, and if it is 2.8 or more, an atherogenic violation of the plasma lipoprotein spectrum is diagnosed. In atherosclerosis, the cholesterol content in the slowly migrating subfractions of o (α-lipoproteins) is also determined and the cholesterol atherogenic index is calculated. With a value of 19.5 or more, atherosclerosis is diagnosed. The sensitivity of the method increases by more than five times as compared with the prototype. . f-ly. (L 4 and SP

Description

 one

The invention relates to medicine, namely to clinical and biochemical laboratory diagnostics and cardiology.

The purpose of the invention is to increase the accuracy of the diagnostic method by increasing its sensitivity.

The method is carried out as follows.

After a 12-hour fasting in the morning on an empty stomach, into a dry clean tube containing crystalline EDTA as an anticoagulant, from the vein of the elbow bend of the test is taken

lo 2 ml of blood. Plasma cholesterol content of high-density lipoproteins () and total cholesterol was determined by the methods of Producing also electrophoretic fractionation of a lipid-stained plasma using the disc electrophoresis of lipoproteins in a multi-layer polyacrylamide gel (D), after exchanging in a buffer solution For one day, the densitometric recording of the disk electrophoregram of lipoproteins (LP) is performed in the glass tubes of the columns of a polyacrylamide gel, for which densities can be used itometer

The densitogram of the disk electrophore- gogram has the following form (from the cathode to the anode): I - albumin, coupled. with non-tertiary fatty acids (NEFA); 2 - rapidly migrating podfra.ktsi with / -lipoproteins; 3 — slowly migrating sub-fraction of poproteins; 4- / 3-lipoproteins; 5 - pre-p lipoproteins; 6 chilomicrons Albumin-NEFA and chylomicrons are not directly related to the formation of atherosclerotic changes in the body, therefore they are not taken into account. In order to quantify diagnostically important peaks, their height and width are measured at mid-height. The values obtained multiply each other and a constant factor of 1.064. The sum of the areas of the peaks is added and taken as 100%. Then the percentage of each peak is determined. In addition, the peak area of the rapidly and slowly migrating subfraction of of lipoproteins is added and taken as the sum of them for 100%, then their percentage and the ratio between the rapidly and slowly migrating subfractions are calculated. After that, the index of ateiogenicity is calculated by the formula ". (

d

IA

g 5

0 5 0 5

five

0

d is the percentage of slowly migrating lipoprotein subfraction.

With an AI of 2.8 and Baud, an atherogenic disorder of the plasma lipoprotein spectrum is diagnosed.

The cholesteric atherogenic index is found as follows. The sum of the two peaks in (-lipoproteins (quickly and slowly migrating subfraction) is taken as the content of al- KS in mmol / l. The ratio of these subfractions and the cholesterol content in them are found. Next, cholesterol level is calculated atherogenic formula

 xs - (xs)

. ™ - oG-xs;

where CHIA is cholesterol, atheroheic index;

CS - total blood plasma cholesterol; o (-XC is the cholesterol content of eLipoproteins; tx -XC is the cholesterol content

slowly migrating subfraction (α-lipoproteins. If the value of a HIA is 19.5 or more, atherosclerosis is diagnosed.

, PRI me R 1. Sick Sh., 43 years old, diagnosed with residual changes after experiencing tuberculosis of triumphal lymph nodes. Segmental pneumosclerosis of the latch segment on the left.

The laboratory was examined prior to treatment. Of the standardized uniform tests, tests for atherogenic disorders in the lipoprotein plasma spectrum were used. Tests for the content of cholesterol, total plasma cholesterol (cholesterol) and cholesterol coefficient of atherogenicity of the lipoprotein spectrum cholesterol of 14.1ХС1 о (-С

Id - atherogenic index;

but. - percentage of pre

 / i-lipoproteins; b is the percentage of p-lipoproteins;

c is the ratio between the rapidly and slowly migrating podraktsi o-li1 opprotenov;

The content with (-XC was 1.36 mmol / l at a rate of 1.235-1.435 with XtSxl values, 335 ± 0.051 mmol / l. Chl content was 5.10 mmol / l at a rate of 3.909-4.382 with X ± Sx4 values, 145 ±

± 0,121 mmol / l. The cholesterol coefficient of atherogenicity of the lipoprotein spectrum is 2.75 at a rate of 1.986 -, 2.406 with Xt values of S.x 2.19610.107.

51

The determination of the atherogenicity of the lipoprotein spectrum by a known method yields a value of 0.59, which differs very little from the normal value (0.645-0.745 with XiS values 0.69510.026), revealing some prevalence of anti-atherogenic lipoproteins - HDL in a patient with pulmonary pathology ,

The determination of the atherogenicity of the lipoprotein spectrum by the proposed method gives the following result: IA 1.055 (at a rate of 1.240-1.816 with values of X ± S 1, 528 ± 0, 147).

Assessing the data obtained, it is impossible not to pay attention to the fact that the indicator found is far from the criteria for the detection of atherosclerosis, which was not revealed even with a careful clinical study of the patient, if the atherogenic index of the lipoprotein spectrum found by a known method is 1.77 times less than the average the atherogenic index of the lipoprotein spectrum determined by the proposed method is 1, 448 times less than the average norm of the norm, the proposed method more correctly reflects the the pathochemical changes in the patient's body than cholesterol CA, since with careful clinical and experimental research, the patient did not reveal atherosclerosis, pulmonary pathologists usually determine the total fraction of (α-lipoproteins.

Example 2: Patient P, 57 years old, diagnosed with primary: cirrhotic tuberculosis in the left lobe of the left lung with cavities, BC and concomitant: coronary artery disease: angina pectoris. Atherosclerotic aorta, coronary and cerebral vessels, pen 1-11 stage.

Laboratory role was investigated prior to treatment. Of the standardized unified tests, tests on the content of atherogenic disorders in the lipoprotein spectrum of blood plasma were used for tests on o (-C, total plasma cholesterol (C) and the cholesterol coefficient of atherogenicity (CA) of the lipoprotein spectrum (-XC),. Of- XC

The content of cf-XC was 0.526 mmold (at a rate of 1.235-1.435 with

 xtS l, 335tO, 051 mmol / l), containing 67516 6

cholesterol was 2, mmol / l (at a rate of 3.908-4.382 with values of 15tO, 121 mmol / l). Choleste, the hemoglobin atherogenic coefficient of the lipoprotein spectrum is 2.865 (at a rate of 986t2.406, with the values of XtS Г2.196 iO, P7).

The determination of the atherogenicity of the lipo10 protein spectrum by a known method yields a value of 0.62, which is practically the same as the normal value (0.645-0.745 with XtS values of 0, 695 0.026).

15 The determination of the atherogenicity of the lipoprotein spectrum by the proposed method yields the following results:, 8 at a rate of 1.240-1.816 with values of X ± S x 1, 528 + 0, 147. The resulting 20 indicator is 1.83 times the average norm, differing by 1.0 units from the upper limit of the norm.

Estimating the data obtained, it is not necessary to note that if the traditional indicator of cholesterol coefficients is C (o (-С))

atherogenicity-j ---- -

O - LS

reflecting the ratio of the content of atherogenic lipoproteins to that of atherogenic drugs, is increased by 1.3 times in this patient (differing by 0.46 units from the upper limit of normal), then the index determined by

25, the proposed method is 1.83 times the average norm, differing by 1.0 units. from the upper limit of the norm. Therefore, the proposed method correctly reflects

The 4D content of biochemical changes in concomitant atherosclerosis is more sensitive and accurate than the known one associated with the determination of the cholesterol coefficient of atherorability of the plasma lipoprotein spectrum.

A known method for assessing atherogenic disorders in the lipoprotein spectrum of plasma in this case turns out to be ineffective.

PRI me R 3. Sick E., 70 years old, diagnosed with the main one: infiltrative pulmonary tuberculosis in the decay phase, BC +, acute // lobar right-sided pneumonia, and related: atherosclerosis of the aorta, coronary and cerebral vessels, atherosclerotic cardiosclerosis, LSN 1-P Art.

Of the standardized unified tests, tests for the levels of cholesterol, total plasma cholesterol (cholesterol) and cholesterol coefficient of atherogenity of the lipoprotein spectrum j were used to judge atherogenic disorders in the lipoproteinogogram blood plasma spectrum.

cx; -hs. . . .

The content was 0.800 mmol / l (at a rate of 1.235-1.435 with values of X ± SV 1, 551 mmol / l), the content of cholesterol was 3.80 mmol / l (at a rate of 3.908-4.382 with values of X ± 8x4, 1 21 mmol / 1 Cholesterol atherogenic coefficient of the lipoprotein spectrum 3.75, (at a rate of 1.986-2.406 with values of Xi S Y 2, 196 t of O, 107).

The determination of the atherogenicity of the lipoprotein spectrum by a known method gives a value of 0.59, which differs very little from the norm (0.645-0.745 with Xt ± S values to 0.695 + 0.026), revealing some prevalence of anti-atherogenic lipoproteins - HDL cholesterol in a patient with combined pathology.

The determination of the atherogenicity of the lnpoprotein spectrum by the proposed method yields the following result: IA 40.44 (at a rate of 1.240-1.816 with values of X ± SY1.52810.147).

Estimating the data obtained, on: b — pay attention to the following: the indicator of cholesterol athephogenicity is 1.7 times (3.75: 2.196 exceeds the average norm, confirming atherogenic changes in the body; the indicator found by the proposed method is 25.5 times (40 , 44: 1, 528) mean the average rate of the norm, which confirms its much higher sensitivity compared to the previous (traditional) one.

The known method of assessing atherogenic disorders in the plasma lipoprotein spectrum in this patient was ineffective.

PRI me R 4. A patient, 67 years old, was admitted with a diagnosis of sypholitis, angina pectoris, rest and stress. Aeroelerotic cardiosclerosis. Atherosclerosis of the aorta, coronary arteries, H ,.

When biochemical ksledovaniya blood plasma by generally accepted methods, it turned out that the content of total

0

Q

Q with

0

five

cholesterol is 5.224 mmol / l cholesterol-HDL cholesterol 0.864 mmol / l. Indicator cholesterol coefficient of atherogenic lipoprotein spectrum:

XC- (l-XC), - -, .- .- equal to 5.046 (at the norm

1.8-3.3 with XtS values of f2.196 ± 0.107).

Additionally, a disc electrophoretic study of the spectrum of lipoproteins in the columns of a polyacrylamide gel was carried out.

Then cholesterol is determined.

v "A XC- ()

atherogenity index of CHIA -7)

lsm

equal to 34.88 (at a rate of 3.253-5.520 with values of X ± S x4.227tO, 107).

Since the resulting index turned out to be more than 19.5, ateroslerosis is diagnosed in the patient.

PRI me R 5. A 70-year-old patient was admitted with a diagnosis of coronary artery disease, angina pectoris diopathy (oterosclerotic) and post-infarction cardiosclerosis, heart attack, atherosclerosis of the coronary arteries, aorta.

Yrie biochemical study of blood plasma by conventional methods turned out that the total cholesterol content is 5.983 mmol / l, o (-XC, 0.7481 mmol / l. Indicator of cholesterol coefficient of atherogenic plasma lipoprotein spectrum, found by a known method and determined by the formula (5 - XC)

equal to 6.997 (at a rate of 1.8-3.3 with values of Xt Sx 2.196t 0.107). I Additionally, a disc electrophoretic study of the spectrum of lipoproteins in the columns of a polyacrylamide gel was carried out.

Next, cholesterol inHC- (e (-XC)) is calculated.

atherogenic deck jvr

"M

 19,533 (19,5).

Since the resulting figure is 19.5, atherosclerosis is diagnosed in this patient.

Thus, the proposed method is more than 5 times more sensitive than the known method.

Claims (2)

1. The method of determining atherogenic disorders of the lipoprotein spectrum 9U67 pa blood plasma by electrophoretic separation of lipoproteins, determining the percentage of npe- / J-lipoproteins, / J-lipoproteins and s (-lipoproteins, followed by calculating the ratio of the sum of pre-; 3-lipoproteins and -lipoproteins with o (-lipoprotein Wherein, in order to increase the accuracy of the method by increasing its sensitivity, C-lipoproteins are further divided into quickly and slowly migrating subfractions, their percentage is determined, and the atherogenic index is calculated by the formulas e YA i2ib) c. - where IA is atherogenic index, a is 20 percent of pre- / y-lipoproteins, b is the percentage of / 5-lipoproteins, c is the ratio between the rapidly and slowly migrating subfractions of lipoproteins, d is 25 percent of the slowly migrating subfractile a -lipoproteigum, and with an index value of 2.8 or more, atherogenic disorder of the plasma-protein-spectrum spectrum is determined 2. The method according to claim 1, in which, in order to improve the accuracy of determination in atherosclerosis, the level of total blood cholesterol, cholesterol "/ -lipoproteins and cholesterol slowly migrating is additionally determined subformations of α-lipoproteins with the subsequent calculation of cholesterol atherogenic index by the formula -..., cholesterol - o (-HS HIL -j-np where HIL - choled-XL „ sterol atherogenic index, plasma cholesterol total cholesterol, of-XC - cholesterol content with α-lipoproteins and atherosclerosis.