Different density and are indicators of lipid metabolism. There are various methods for the quantitative determination of total lipids: colorimetric, nephelometric.

The principle of the method. The hydrolysis products of unsaturated lipids form a red compound with the phosphovaniline reagent, the color intensity of which is directly proportional to the content of total lipids.

Most lipids are found in the blood not in a free state, but as part of protein-lipid complexes: chylomicrons, α-lipoproteins, β-lipoproteins. Lipoproteins can be separated by various methods: centrifugation in saline solutions of various densities, electrophoresis, thin layer chromatography. During ultracentrifugation, chylomicrons and lipoproteins of different density are isolated: high (HDL - α-lipoproteins), low (LDL - β-lipoproteins), very low (VLDL - pre-β-lipoproteins), etc.

Fractions of lipoproteins differ in the amount of protein, the relative molecular weight of lipoproteins, and the percentage of individual lipid components. Thus, α-lipoproteins containing a large amount of protein (50-60%) have a higher relative density (1.063-1.21), while β-lipoproteins and pre-β-lipoproteins contain less protein and a significant amount of lipids - up to 95% of the total relative molecular weight and low relative density (1.01-1.063).

Method principle. When LDL of blood serum interacts with a heparin reagent, turbidity appears, the intensity of which is determined photometrically. The heparin reagent is a mixture of heparin and calcium chloride.

Material under study: blood serum.

Reagents: 0.27% CaCl 2 solution, 1% heparin solution.

Equipment: micropipette, FEK, cuvette with an optical path length of 5 mm, test tubes.

PROGRESS. 2 ml of a 0.27% solution of CaCl 2 and 0.2 ml of blood serum are added to the test tube, mixed. Determine the optical density of the solution (E 1) against a 0.27% CaCl 2 solution in cuvettes with a red light filter (630 nm). The solution from the cuvette is poured into a test tube, 0.04 ml of a 1% heparin solution is added with a micropipette, mixed, and exactly after 4 minutes the optical density of the solution (E 2) is again determined under the same conditions.

The difference in optical density is calculated and multiplied by 1000 - the empirical coefficient proposed by Ledvina, since the construction of a calibration curve is associated with a number of difficulties. The answer is expressed in g/l.

x (g / l) \u003d (E 2 - E 1) 1000.

. The content of LDL (b-lipoproteins) in the blood varies depending on age, gender and is normally 3.0-4.5 g / l. An increase in the concentration of LDL is observed in atherosclerosis, obstructive jaundice, acute hepatitis, chronic liver diseases, diabetes, glycogenosis, xanthomatosis and obesity, a decrease in b-plasmocytoma. The average cholesterol content in LDL is about 47%.

Determination of total cholesterol in blood serum based on the Liebermann-Burchard reaction (Ilk method)

Exogenous cholesterol in the amount of 0.3-0.5 g comes with food, and endogenous cholesterol is synthesized in the body in the amount of 0.8-2 g per day. Especially a lot of cholesterol is synthesized in the liver, kidneys, adrenal glands, arterial wall. Cholesterol is synthesized from 18 molecules of acetyl-CoA, 14 molecules of NADPH, 18 molecules of ATP.

When acetic anhydride and concentrated sulfuric acid are added to the blood serum, the liquid turns red, blue, and finally green. The reaction is due to the formation of green sulfonic acid cholesterylene.

Reagents: Liebermann-Burchard reagent (a mixture of glacial acetic acid, acetic anhydride and concentrated sulfuric acid in a ratio of 1:5:1), standard (1.8 g / l) cholesterol solution.

Equipment: dry test tubes, dry pipettes, FEK, cuvettes with an optical path length of 5 mm, a thermostat.

PROGRESS. All test tubes, pipettes, cuvettes must be dry. It is necessary to work with the Liebermann-Burchard reagent very carefully. 2.1 ml of the Liebermann-Burchard reagent is placed in a dry tube, 0.1 ml of non-hemolyzed blood serum is added very slowly along the wall of the tube, the tube is vigorously shaken, and then thermostated for 20 minutes at 37ºС. An emerald green color develops, which is colorimetric on FEC with a red light filter (630-690 nm) against the Liebermann-Burchard reagent. The optical density obtained on the FEC is used to determine the concentration of cholesterol according to the calibration curve. The found concentration of cholesterol is multiplied by 1000, since 0.1 ml of serum is taken in the experiment. The conversion factor to SI units (mmol/l) is 0.0258. The normal content of total cholesterol (free and esterified) in the blood serum is 2.97-8.79 mmol / l (115-340 mg%).

Construction of a calibration graph. From a standard solution of cholesterol, where 1 ml contains 1.8 mg of cholesterol, take 0.05; 0.1; 0.15; 0.2; 0.25 ml and adjusted to a volume of 2.2 ml with the Liebermann-Burchard reagent (respectively 2.15; 2.1; 2.05; 2.0; 1.95 ml). The amount of cholesterol in the sample is 0.09; 0.18; 0.27; 0.36; 0.45 mg. The obtained standard solutions of cholesterol, as well as experimental test tubes, are vigorously shaken and placed in a thermostat for 20 minutes, after which they are photometered. The calibration graph is built according to the extinction values ​​obtained as a result of photometry of standard solutions.

Clinical and diagnostic value. In violation of fat metabolism, cholesterol can accumulate in the blood. An increase in blood cholesterol (hypercholesterolemia) is observed in atherosclerosis, diabetes mellitus, obstructive jaundice, nephritis, nephrosis (especially lipoid nephrosis), and hypothyroidism. A decrease in blood cholesterol (hypocholesterolemia) is observed with anemia, starvation, tuberculosis, hyperthyroidism, cancer cachexia, parenchymal jaundice, CNS damage, febrile conditions, with the introduction

lipids called fats that enter the body with food and are formed in the liver. Blood (plasma or serum) contains 3 main classes of lipids: triglycerides (TG), cholesterol (CS) and its esters, phospholipids (PL).
Lipids are able to attract water, but most of them do not dissolve in the blood. They are transported in a protein-bound state (in the form of lipoproteins or, in other words, lipoproteins). Lipoproteins differ not only in composition, but also in size and density, but their structure is almost the same. The central part (core) is represented by cholesterol and its esters, fatty acids, triglycerides. The shell of the molecule consists of proteins (apoproteins) and water-soluble lipids (phospholipids and non-esterified cholesterol). The outer part of apoproteins is able to form hydrogen bonds with water molecules. Thus, lipoproteins can partially dissolve in fats, partially in water.
Chylomicrons after entering the blood break down into glycerol and fatty acids, resulting in the formation of lipoproteins. Cholesterol-containing residues of chylomicrons are processed in the liver.
From cholesterol and triglycerides in the liver, very low density lipoproteins (VLDL) are formed, which donate part of the triglycerides to peripheral tissues, while their remains return to the liver and are converted into low density lipoproteins (LDL).
LPN II are transporters of cholesterol for peripheral tissues, which is used to build cell membranes and metabolic reactions. In this case, non-esterified cholesterol enters the blood plasma and binds to high-density lipoproteins (HDL). Esterified cholesterol (associated with esters) is converted to VLDL. Then the cycle repeats.
The blood also contains intermediate density lipoproteins (LDL), which are the remains of chylomicrons and VLDL and contain large amounts of cholesterol. LDL in liver cells with the participation of lipase are converted into LDL.
Blood plasma contains 3.5-8 g/l of lipids. An increase in the level of lipids in the blood is called hyperlipidemia, and a decrease is called hypolipidemia. The indicator of total blood lipids does not give a detailed idea of ​​the state of fat metabolism in the body.
Diagnostic value is the quantitative determination of specific lipids. The lipid composition of blood plasma is presented in the table.

Lipid composition of blood plasma

Fraction of lipids		Norm indicator
General lipids		4.6-10.4 mmol/l
Phospholipids		1.95-4.9 mmol/l
Lipid phosphorus		1.97-4.68 mmol/l
Neutral Fats		0-200 mg%
Triglycerides		0.565-1.695 mmol/l (serum)
Non-esterified fatty acids		400-800 mmol/l
Free fatty acids		0.3-0.8 µmol/l
Total cholesterol (there are age norms)		3.9-6.5 mmol/l (unified method)
free cholesterol		1.04-2.33 mmol/l
Cholesterol esters		2.33-3.49 mmol/l
HDL	M	1.25-4.25 g/l
	F	2.5-6.5 g/l
LDL		3-4.5 g/l

A change in the lipid composition of the blood - dyslipidemia - is an important sign of atherosclerosis or a condition preceding it. Atherosclerosis, in turn, is the main cause of coronary heart disease and its acute forms (angina pectoris and myocardial infarction).
Dyslipidemias are divided into primary, associated with congenital metabolic disorders, and secondary. The causes of secondary dyslipidemia are physical inactivity and overnutrition, alcoholism, diabetes mellitus, hyperthyroidism, cirrhosis of the liver, and chronic renal failure. In addition, they can develop during treatment with glucocorticosteroids, B-blockers, progestins and estrogens. The classification of dyslipidemia is presented in the table.

Classification of dyslipidemias

A type	An increase in blood levels
	Lipoprotein	lipids
I	Chylomicrons	Cholesterol, triglycerides
On the	LDL	Cholesterol (not always)
A type	An increase in blood levels
	Lipoprotein	lipids
Nb	LDL, VLDL	Cholesterol, triglycerides
III	VLDL, LPPP	Cholesterol, triglycerides
IV	VLDL	Cholesterol (not always), triglycerides
V	Chylomicrons, VLDL	Cholesterol, triglycerides

Lipids are a group of low molecular weight substances characterized by different solubility in organic solvents and insoluble in water. Lipids in the blood are mainly in the form of chylomicrons and lipoproteins. There are three main classes of lipids in blood plasma: cholesterol and its esters, triglycerides (neutral fats), and phospholipids.

An increase in total lipids in the blood serum is called hyperlidemia. It is observed after eating - this is a physiological phenomenon (alimentary hyperlipidemia). Physiological hyperlipidemia occurs 1-4 hours after a meal. The increase in blood lipids after eating is the higher, the lower the level of lipids in the blood on an empty stomach.

The study of total lipids gives an approximate idea of ​​the state of lipid metabolism in the subject.

An increase in blood lipids may be accompanied by the following diseases:

Acute and chronic hepatitis, obstructive jaundice. However, with the most severe
lesions of the liver parenchyma, the content of lipids in the blood decreases (mechanical
jaundices are also accompanied by hyperlipidemia);

Diabetes mellitus is accompanied by severe hyperlipemia, which, as a rule,
develops in parallel with acidosis. Hyperlipemia in diabetes is caused by increased
mobilization of fat from fat depots and delivery of lipids to the liver. Such is the nature
hyperlipidemia and pancreatitis;

Some kidney diseases. In acute and chronic nephritis without edema, the number of
blood lipid levels are normal, with edema - increased. With lipoid nephrosis
the amount of lipids increases by 2-6 times [Pokrovsky A.A., 1969];

The so-called spontaneous hyperlipemia is a rare hereditary disease, on
observed mainly in men. The basis of the disease is a violation of the transition
yes lipids from blood to tissues due to lack of tissue lipases. In persons suffering from this
pathology, there is a pronounced tendency to the development of atherosclerosis.

Currently, the study of total lipids is practically not used in clinical practice due to the low information content of this indicator.

Serum triglycerides

Triglycerides (TG), or neutral fats, are esters of the triatomic alcohol glycerol and higher fatty acids. TG enter the body with food (exogenous TG) and are synthesized in the body (endogenous TG). The latter are formed in the liver mainly from carbohydrates. TG are the main form of accumulation of fatty acids in the body and the main source of energy in humans. Normal concentrations of TG in serum are presented in table. 4.22.

In clinical practice, the content of TG in the blood is determined mainly for the detection and typing of dyslipoproteinemia.

tables	a "	1.22. The content of TG in serum is normal	[Titz W., 1986]
			Content	serum TG
Age, years			mg/dl		mmol/l
		men		women		men	women
0-5		30-86		32-99		0,34-0,97	0,36-1,12
6-11		31-108		35-114		0,35-1,22	0,40-1,29
12-15		36-138		41-138		0,41-1,56	0,46-1,56
16-19		40-163		40-128		0,45-1,84	0,45-1,45
20-29		44-185		40-128		0,50-2,09	0,45-1,45
30-39		49-284		38-160		0,55-3,21	0,43-1,81
40-49		56-298		44-186		0,63-3,37	0,50-2,10
50-59		62-288		55-247		0,70-3,25	0,62-2,79
	In persons older	60 years of meaning	decrease slightly

com pancreatitis, chronic renal failure, hypertension, acute myocardial infarction, pregnancy, chronic ischemic heart disease, cerebral vascular thrombosis, hypothyroidism, diabetes mellitus, gout, glycogenosis I, III and VI types, respiratory distress syndrome, thalassemia major, Down's syndrome, Werner's syndrome, anorexia nervosa, idiopathic hypercalcemia, acute intermittent porphyria.

Elevated levels of TG in the blood is a risk factor for the development of coronary artery disease. At the same time, an increase in the level of triglycerides in the blood up to 200-500 mg / dl, or 2.3-5.6 mmol / l, is regarded as severe hypertriglyceridemia, and more than 500 mg / dl, or more than 5.6 mmol / l, as severe hypertriglyceridemia [Dolgov V. et al., 1995].

Pyruvic acid in the blood

Clinical and diagnostic significance of the study

Norm: 0.05-0.10 mmol / l in the blood serum of adults.

PVC content increases in hypoxic conditions caused by severe cardiovascular, pulmonary, cardiorespiratory insufficiency, anemia, malignant neoplasms, acute hepatitis and other liver diseases (most pronounced in the terminal stages of liver cirrhosis), toxicosis, insulin-dependent diabetes mellitus, diabetic ketoacidosis, respiratory alkalosis, uremia , hepatocerebral dystrophy, hyperfunction of the pituitary-adrenal and sympathetic-adrenal systems, as well as the introduction of camphor, strychnine, adrenaline and during heavy physical exertion, tetany, convulsions (with epilepsy).

Clinical and diagnostic significance of determining the content of lactic acid in the blood

Lactic acid(MK) is the end product of glycolysis and glycogenolysis. A significant amount is formed in muscles. From the muscle tissue, MK with the blood flow enters the liver, where it is used for the synthesis of glycogen. At the same time, part of the lactic acid from the blood is absorbed by the heart muscle, which utilizes it as an energy material.

Blood UA level increases with hypoxic conditions, acute purulent inflammatory tissue damage, acute hepatitis, cirrhosis of the liver, renal failure, malignant neoplasms, diabetes mellitus (approximately 50% of patients), mild uremia, infections (especially pyelonephritis), acute septic endocarditis, poliomyelitis, severe diseases blood vessels, leukemia, intense and prolonged muscle exertion, epilepsy, tetany, tetanus, convulsive conditions, hyperventilation, pregnancy (in the third trimester).

Lipids are chemically diverse substances that have a number of common physical, physicochemical and biological properties. Οʜᴎ are characterized by the ability to dissolve in ether, chloroform, other fatty solvents and only slightly (and not always) in water, and also form the main structural component of living cells together with proteins and carbohydrates. The inherent properties of lipids are determined by the characteristic features of the structure of their molecules.

The role of lipids in the body is very diverse. Some of them serve as a form of deposition (triacylglycerols, TG) and transport (free fatty acids - FFA) of substances, during the decay of which a large amount of energy is released, others are the most important structural components of cell membranes (free cholesterol and phospholipids). Lipids take part in the processes of thermoregulation, protection of vital organs (for example, kidneys) from mechanical influences (injuries), protein loss, in creating elasticity of the skin, protecting them from excessive moisture removal.

Some of the lipids are biologically active substances that have the properties of modulators of hormonal influence (prostaglandins) and vitamins (fatty polyunsaturated acids). Moreover, lipids promote the absorption of fat-soluble vitamins A, D, E, K; act as antioxidants (vitamins A, E), largely regulating the process of free radical oxidation of physiologically important compounds; determine the permeability of cell membranes in relation to ions and organic compounds.

Lipids serve as precursors for a number of steroids with a pronounced biological effect - bile acids, vitamins of group D, sex hormones, hormones of the adrenal cortex.

The concept of "total lipids" of plasma includes neutral fats (triacylglycerols), their phosphorylated derivatives (phospholipids), free and ester-bound cholesterol, glycolipids, non-esterified (free) fatty acids.

Clinical and diagnostic value determination of the level of total lipids in plasma (serum) of blood

The norm is 4.0-8.0 g / l.

Hyperlipidemia (hyperlipemia) - an increase in the concentration of total plasma lipids as a physiological phenomenon can be observed 1.5 hours after a meal. Alimentary hyperlipemia is more pronounced, the lower the level of lipids in the patient's blood on an empty stomach.

The concentration of lipids in the blood changes in a number of pathological conditions. So, in patients with diabetes, along with hyperglycemia, there is a pronounced hyperlipemia (often up to 10.0-20.0 g / l). With nephrotic syndrome, especially lipoid nephrosis, the content of lipids in the blood can reach even higher figures - 10.0-50.0 g / l.

Hyperlipemia is a constant phenomenon in patients with biliary cirrhosis of the liver and in patients with acute hepatitis (especially in the icteric period). Elevated blood lipids are usually found in individuals suffering from acute or chronic nephritis, especially if the disease is accompanied by edema (due to the accumulation of plasma LDL and VLDL).

The pathophysiological mechanisms that cause shifts in the content of all fractions of total lipids determine, to a greater or lesser extent, a pronounced change in the concentration of its constituent subfractions: cholesterol, total phospholipids and triacylglycerols.

Clinical and diagnostic significance of the study of cholesterol (CS) in serum (plasma) of blood

The study of the level of cholesterol in the serum (plasma) of the blood does not provide accurate diagnostic information about a specific disease, but only reflects the pathology of lipid metabolism in the body.

According to epidemiological studies, the upper level of cholesterol in the blood plasma of practically healthy people aged 20-29 years is 5.17 mmol/l.

In blood plasma, cholesterol is found mainly in the composition of LDL and VLDL, and 60-70% of it is in the form of esters (bound cholesterol), and 30-40% is in the form of free, non-esterified cholesterol. Bound and free cholesterol make up the amount of total cholesterol.

A high risk of developing coronary atherosclerosis in people aged 30-39 and older than 40 years occurs at cholesterol levels exceeding 5.20 and 5.70 mmol / l, respectively.

Hypercholesterolemia is the most proven risk factor for coronary atherosclerosis. This has been confirmed by numerous epidemiological and clinical studies that have established a link between hypercholesterolemia and coronary atherosclerosis, the incidence of coronary artery disease and myocardial infarction.

The highest level of cholesterol is observed in genetic disorders in the metabolism of LP: familial homo-heterozygous hypercholesterolemia, familial combined hyperlipidemia, polygenic hypercholesterolemia.

In a number of pathological conditions, secondary hypercholesterolemia develops. . It is observed in liver diseases, kidney damage, malignant tumors of the pancreas and prostate, gout, coronary artery disease, acute myocardial infarction, hypertension, endocrine disorders, chronic alcoholism, type I glycogenosis, obesity (in 50-80% of cases).

A decrease in plasma cholesterol levels is observed in patients with malnutrition, with damage to the central nervous system, mental retardation, chronic insufficiency of the cardiovascular system, cachexia, hyperthyroidism, acute infectious diseases, acute pancreatitis, acute purulent-inflammatory processes in soft tissues, febrile conditions, pulmonary tuberculosis, pneumonia, respiratory sarcoidosis, bronchitis, anemia, hemolytic jaundice, acute hepatitis, malignant liver tumors, rheumatism.

Of great diagnostic importance is the determination of the fractional composition of blood plasma cholesterol and its individual LP (primarily HDL) for judging the functional state of the liver. According to the modern concept, the esterification of free cholesterol in HDL is carried out in the blood plasma due to the enzyme lecithin-cholesterol-acyltransferase, which is formed in the liver (this is an organ-specific liver enzyme). The activator of this enzyme is one of the basic components of HDL - apo - Al, which is constantly synthesized in the liver.

Albumin, also produced by hepatocytes, serves as a nonspecific activator of the plasma cholesterol esterification system. This process primarily reflects the functional state of the liver. If the normal coefficient of cholesterol esterification (ᴛ.ᴇ. the ratio of the content of ester-bound cholesterol to total) is 0.6-0.8 (or 60-80%), then in acute hepatitis, exacerbation of chronic hepatitis, cirrhosis of the liver, obstructive jaundice , as well as chronic alcoholism, it decreases. A sharp decrease in the severity of the process of cholesterol esterification indicates a lack of liver function.

Clinical and diagnostic significance of the study of the concentration of total phospholipids in blood serum.

Phospholipids (PL) are a group of lipids containing, in addition to phosphoric acid (as an essential component), an alcohol (usually glycerol), fatty acid residues, and nitrogenous bases. Given the dependence on the nature of alcohol, PL is divided into phosphoglycerides, phosphosphingosines, and phosphoinositides.

The level of total PL (lipid phosphorus) in the blood serum (plasma) is increased in patients with primary and secondary hyperlipoproteinemia types IIa and IIb. This increase is most pronounced in type I glycogenosis, cholestasis, obstructive jaundice, alcoholic and biliary cirrhosis, viral hepatitis (mild course), renal coma, posthemorrhagic anemia, chronic pancreatitis, severe diabetes mellitus, nephrotic syndrome.

For the diagnosis of a number of diseases, it is more informative to study the fractional composition of blood serum phospholipids. For this purpose, thin-layer lipid chromatography methods have been widely used in recent years.

Composition and properties of blood plasma lipoproteins

Almost all plasma lipids are associated with proteins, which gives them good solubility in water. These lipid-protein complexes are commonly referred to as lipoproteins.

According to the modern concept, lipoproteins are high-molecular water-soluble particles, which are complexes of proteins (apoproteins) and lipids formed by weak, non-covalent bonds, in which polar lipids (PL, CXC) and proteins (“apo”) make up the surface hydrophilic monomolecular layer surrounding and protecting the internal phase (consisting mainly of ECS, TG) from water.

In other words, LP are peculiar globules, inside of which there is a fat drop, a core (formed mainly by non-polar compounds, mainly triacylglycerols and cholesterol esters), delimited from water by a surface layer of protein, phospholipids and free cholesterol.

The physical features of lipoproteins (their size, molecular weight, density), as well as the manifestations of physicochemical, chemical and biological properties, largely depend, on the one hand, on the ratio between the protein and lipid components of these particles, on the other hand, on the composition of the protein and lipid components, ᴛ.ᴇ. their nature.

The largest particles, consisting of 98% lipids and a very small (about 2%) proportion of protein, are chylomicrons (XM). Οʜᴎ are formed in the cells of the mucous membrane of the small intestine and are a transport form for neutral dietary fats, ᴛ.ᴇ. exogenous TG.

Table 7.3 Composition and some properties of blood serum lipoproteins (Komarov F.I., Korovkin B.F., 2000)

Criteria for evaluating individual classes of lipoproteins	HDL (alpha-LP)	LDL (beta-LP)	VLDL (pre-beta-LP)	HM
Density, kg/l	1,063-1,21	1,01-1,063	1,01-0,93	0,93
Molecular weight of LP, kD	180-380		3000- 128 000	-
Particle size, nm	7,0-13,0	15,0-28,0	30,0-70,0	500,0 - 800,0
Total proteins, %	50-57	21-22	5-12
Total lipids, %	43-50	78-79	88-95
Free cholesterol, %	2-3	8-10	3-5
Esterified cholesterol, %	19-20	36-37	10-13	4-5
Phospholipids, %	22-24	20-22	13-20	4-7
Triacylglycerols, %
4-8	11-12	50-60	84-87

If exogenous TG are transferred into the blood by chylomicrons, then the transport form endogenous TG are VLDL. Their formation is a protective reaction of the body, aimed at preventing fatty infiltration, and subsequently liver dystrophy.

The dimensions of VLDL are on average 10 times smaller than the size of CM (individual particles of VLDL are 30-40 times smaller than CM particles). They contain 90% of lipids, among which more than half of the content is TG. 10% of total plasma cholesterol is carried by VLDL. Due to the content of a large amount of TG VLDL, an insignificant density is detected (less than 1.0). Determined that LDL and VLDL contain 2/3 (60%) of the total cholesterol plasma, while 1/3 is accounted for by HDL.

HDL- the most dense lipid-protein complexes, since the protein content in them is about 50% of the particle mass. Their lipid component consists half of phospholipids, half of cholesterol, mainly ester-bound. HDL is also constantly formed in the liver and partly in the intestine, as well as in the blood plasma as a result of the “degradation” of VLDL.

If LDL and VLDL deliver cholesterol from the liver to other tissues(peripheral), including vascular wall, then HDL transport cholesterol from cell membranes (primarily the vascular wall) to the liver. In the liver, it goes to the formation of bile acids. In accordance with such participation in cholesterol metabolism, VLDL and themselves LDL are called atherogenic, a HDL– antiatherogenic drugs. Under atherogenicity, it is customary to understand the ability of lipid-protein complexes to contribute (transfer) free cholesterol contained in LP into tissues.

HDL compete for cell membrane receptors with LDL, thereby counteracting the utilization of atherogenic lipoproteins. Since the surface monolayer of HDL contains a large amount of phospholipids, favorable conditions are created at the point of contact of the particle with the outer membrane of endothelial, smooth muscle and any other cells for the transfer of excess free cholesterol to HDL.

At the same time, the latter lingers in the surface monolayer of HDL only for a very short time, since it undergoes esterification with the participation of the LCAT enzyme. The formed ECS, being a non-polar substance, moves into the internal lipid phase, freeing vacancies for repeating the act of capturing a new CXC molecule from the cell membrane. From here: the higher the activity of LCAT, the more effective the anti-atherogenic effect of HDL, which are considered as LCAT activators.

If the balance between the influx of lipids (cholesterol) into the vascular wall and their outflow from it is disturbed, conditions are created for the formation of lipoidosis, the most famous manifestation of which is atherosclerosis.

In accordance with the ABC nomenclature of lipoproteins, primary and secondary lipoproteins are distinguished. Primary LPs are formed by any one apoprotein by chemical nature. They are conventionally classified as LDL, which contain about 95% of apoprotein-B. All the rest are secondary lipoproteins, which are associated complexes of apoproteins.

Normally, approximately 70% of plasma cholesterol is in the composition of "atherogenic" LDL and VLDL, while about 30% circulates in the composition of "anti-atherogenic" HDL. With this ratio in the vascular wall (and other tissues), the balance of the rates of inflow and outflow of cholesterol is maintained. This determines the numerical value cholesterol coefficient atherogenicity, which, with the indicated lipoprotein distribution of total cholesterol 2,33 (70/30).

According to the results of mass, epidemiological observations, at a concentration of total cholesterol in plasma of 5.2 mmol/l, a zero balance of cholesterol in the vascular wall is maintained. An increase in the level of total cholesterol in the blood plasma of more than 5.2 mmol / l leads to its gradual deposition in the vessels, and at a concentration of 4.16-4.68 mmol / l, a negative balance of cholesterol in the vascular wall is observed. The level of total plasma (serum) cholesterol in excess of 5.2 mmol / l is considered pathological.

Table 7.4 Scale for assessing the likelihood of developing coronary artery disease and other manifestations of atherosclerosis

(Komarov F.I., Korovkin B.F., 2000)

Hyperlipidemia (hyperlipemia) - an increase in the concentration of total plasma lipids as a physiological phenomenon can be observed 1-4 hours after a meal. Alimentary hyperlipemia is more pronounced, the lower the level of lipids in the patient's blood on an empty stomach.

The concentration of lipids in the blood changes in a number of pathological conditions:

Nephrotic syndrome, lipoid nephrosis, acute and chronic nephritis;

Biliary cirrhosis of the liver, acute hepatitis;

Obesity - atherosclerosis;

Hypothyroidism;

Pancreatitis, etc.

The study of the level of cholesterol (CS) reflects only the pathology of lipid metabolism in the body. Hypercholesterolemia is a documented risk factor for coronary atherosclerosis. CS is an essential component of the membrane of all cells, the special physicochemical properties of CS crystals and the conformation of its molecules contribute to the orderliness and mobility of phospholipids in membranes with temperature changes, which allows the membrane to be in an intermediate phase state (“gel-liquid crystal”) and maintain physiological functions . CS is used as a precursor in the biosynthesis of steroid hormones (gluco- and mineralocorticoids, sex hormones), vitamin D 3 , and bile acids. It is conditionally possible to distinguish 3 pools of CS:

A - rapidly exchanging (30 g);

B - slowly exchanging (50 g);

B - very slowly exchanging (60 g).

Endogenous cholesterol is synthesized in a significant amount in the liver (80%). Exogenous cholesterol enters the body in the composition of animal products. Transport of cholesterol from the liver to extrahepatic tissues is carried out

LDL. Excretion of cholesterol from the liver from extrahepatic tissues to the liver is produced by mature forms of HDL (50% LDL, 25% HDL, 17% VLDL, 5% HM).

Hyperlipoproteinemia and hypercholesterolemia (Fredrickson classification):

type 1 - hyperchylomicronemia;

type 2 - a - hyper-β-lipoproteinemia, b - hyper-β and hyperpre-β-lipoproteinemia;

type 3 - dis-β-lipoproteinemia;

type 4 - hyper-pre-β-lipoproteinemia;

Type 5 - hyper-pre-β-lipoproteinemia and hyperchylomicronemia.

The most atherogenic are types 2 and 3.

Phospholipids - a group of lipids containing, in addition to phosphoric acid (an obligatory component), alcohol (usually glycerol), fatty acid residues and nitrogenous bases. In clinical and laboratory practice, there is a method for determining the level of total phospholipids, the level of which increases in patients with primary and secondary hyperlipoproteinemia IIa and IIb. The decrease occurs in a number of diseases:

Alimentary dystrophy;

fatty degeneration of the liver,

portal cirrhosis;

Progression of atherosclerosis;

Hyperthyroidism, etc.

Lipid peroxidation (LPO) is a free-radical process, the initiation of which occurs during the formation of reactive oxygen species - superoxide O 2 . ; hydroxyl radical HO . ; hydroperoxide radical HO 2 . ; singlet oxygen O 2 ; hypochlorite ion ClO - . The main substrates of lipid peroxidation are polyunsaturated fatty acids that are in the structure of membrane phospholipids. Iron metal ions are the strongest catalyst. LPO is a physiological process that is important for the body, as it regulates membrane permeability, affects cell division and growth, starts phagosynthesis, and is a pathway for the biosynthesis of certain biological substances (prostaglandins, thromboxanes). The LPO level is controlled by the antioxidant system (ascorbic acid, uric acid, β-carotene, etc.). The loss of balance between the two systems leads to the death of cells and cellular structures.

For diagnostics, it is customary to determine the content of lipid peroxidation products in plasma and erythrocytes (diene conjugates, malondialdehyde, Schiff bases), the concentration of the main natural antioxidant - alpha-tocopherol with the calculation of the MDA / TF coefficient. An integral test for assessing lipid peroxidation is the determination of the permeability of erythrocyte membranes.

2. pigment exchange a set of complex transformations of various colored substances in the human and animal body.

The most well-known blood pigment is hemoglobin (chromoprotein, which consists of the protein part of globin and the prosthetic group, represented by 4 hemes, each heme consists of 4 pyrrole nuclei, which are interconnected by methine bridges, in the center is an iron ion with an oxidation state of 2 +) . The average life span of an erythrocyte is 100-110 days. At the end of this period, the destruction and destruction of hemoglobin occurs. The decay process begins already in the vascular bed, ends in the cellular elements of the system of phagocytic mononuclear cells (Kupffer cells of the liver, histiocytes of the connective tissue, plasma cells of the bone marrow). Hemoglobin in the vascular bed binds to plasma haptoglobin and is retained in the vascular bed without passing through the renal filter. Due to the trypsin-like action of the haptoglobin beta chain and the conformational changes caused by its influence in the heme porphyrin ring, conditions are created for easier destruction of hemoglobin in the cellular elements of the phagocytic mononuclearon system. The high-molecular green pigment thus formed verdoglobin(synonyms: verdohemoglobin, choleglobin, pseudohemoglobin) is a complex consisting of globin, a broken porphyrin ring system and ferric iron. Further transformations lead to the loss of iron and globin by verdoglobin, as a result of which the porphyrin ring unfolds into a chain and a low molecular weight green bile pigment is formed - biliverdin. Almost all of it is enzymatically reduced to the most important red-yellow bile pigment - bilirubin, which is a common component of blood plasma. On the surface of the plasma membrane of the hepatocyte undergoes dissociation. In this case, the released bilirubin forms a temporary associate with the lipids of the plasma membrane and moves through it due to the activity of certain enzyme systems. Further passage of free bilirubin into the cell occurs with the participation of two carrier proteins in this process: ligandin (it transports the main amount of bilirubin) and protein Z.

Ligandin and protein Z are also found in the kidneys and intestines, therefore, in case of liver failure, they are free to compensate for the weakening of detoxification processes in this organ. Both of them are quite well soluble in water, but lack the ability to move through the lipid layer of the membrane. Due to the binding of bilirubin to glucuronic acid, the inherent toxicity of free bilirubin is largely lost. Hydrophobic, lipophilic free bilirubin, easily dissolving in membrane lipids and penetrating as a result into mitochondria, uncouples respiration and oxidative phosphorylation in them, disrupts protein synthesis, the flow of potassium ions through the membrane of cells and organelles. This negatively affects the state of the central nervous system, causing a number of characteristic neurological symptoms in patients.

Bilirubinglucuronides (or bound, conjugated bilirubin), in contrast to free bilirubin, immediately react with a diazoreactive (“direct” bilirubin). It should be borne in mind that in the blood plasma itself, bilirubin that is not conjugated with glucuronic acid can either be associated with albumin or not. The last fraction (not associated with albumin, lipids, or other blood components of bilirubin) is the most toxic.

Bilirubinglucuronides, thanks to the enzyme systems of the membranes, actively move through them (against the concentration gradient) into the bile ducts, being released along with the bile into the intestinal lumen. In it, under the influence of enzymes produced by the intestinal microflora, the glucuronide bond is broken. The released free bilirubin is restored with the formation in the small intestine, first mesobilirubin, and then mesobilinogen (urobilinogen). Normally, a certain part of mesobilinogen, being absorbed in the small intestine and in the upper part of the large intestine, enters the liver through the portal vein system, where it is almost completely destroyed (by oxidation), turning into dipyrrole compounds - propent-diopent and mesobilileucan.

Mesobilinogen (urobilinogen) does not enter the general circulation. Part of it, together with the products of destruction, is again sent to the intestinal lumen as part of bile (enterohepotal circulation). However, even with the most minor changes in the liver, its barrier function is largely “removed” and mesobilinogen first enters the general circulation and then into the urine. The bulk of it is sent from the small intestine to the large intestine, where, under the influence of anaerobic microflora (E. coli and other bacteria), it undergoes further restoration with the formation of stercobilinogen. The resulting stercobilinogen (daily amount of 100-200 mg) is almost completely excreted in the feces. In the air, it oxidizes and turns into stercobilin, which is one of the fecal pigments. A small part of stercobilinogen is absorbed through the mucous membrane of the large intestine into the system of the inferior vena cava, delivered with blood to the kidneys and excreted in the urine.

Thus, in the urine of a healthy person, mesobilinogen (urobilinogen) is absent, but it contains some stercobilin (which is often incorrectly called “urobilin”)

To determine the content of bilirubin in the serum (plasma) of the blood, mainly chemical and physico-chemical research methods are used, among which there are colorimetric, spectrophotometric (manual and automated), chromatographic, fluorimetric and some others.

One of the important subjective signs of a violation of pigment metabolism is the appearance of jaundice, which is usually noted when the level of bilirubin in the blood is 27-34 μmol / l or more. The causes of hyperbilirubinemia can be: 1) increased hemolysis of erythrocytes (more than 80% of total bilirubin is represented by unconjugated pigment); 2) violation of the function of the liver cells and 3) a delay in the outflow of bile (hyperbilirubinemia is of hepatic origin, if more than 80% of total bilirubin is conjugated bilirubin). In the first case, they talk about the so-called hemolytic jaundice, in the second - about parenchymal (may be caused by hereditary defects in the processes of bilirubin transport and its glucuronidation), in the third - about mechanical (or obstructive, congestive) jaundice.

With parenchymal jaundice there are destructive-dystrophic changes in the parenchymal cells of the liver and infiltrative changes in the stroma, leading to an increase in pressure in the bile ducts. Stagnation of bilirubin in the liver is also facilitated by a sharp weakening of metabolic processes in the affected hepatocytes, which lose the ability to normally perform various biochemical and physiological processes, in particular, transfer bound bilirubin from cells into bile against a concentration gradient. An increase in the concentration of conjugated bilirubin in the blood leads to its appearance in the urine.

The most “subtle” sign of liver damage in hepatitis is the appearance mesobilinogen(urobilinogen) in the urine.

With parenchymal jaundice, the concentration of conjugated (conjugated) bilirubin in the blood increases mainly. The content of free bilirubin increases, but to a lesser extent.

At the heart of the pathogenesis of obstructive jaundice is the cessation of the flow of bile into the intestine, which leads to the disappearance of stercobilinogen from the urine. With congestive jaundice, mainly the content of conjugated bilirubin in the blood increases. Extrahepatic cholestatic jaundice is accompanied by a triad of clinical signs: discolored feces, dark urine, and itchy skin. Intrahepatic cholestasis is clinically manifested by skin itching and jaundice. In a laboratory study, hyperbilirubinemia (due to associated), bilirubinuria, an increase in alkaline phosphatase with normal values ​​of transaminases in the blood serum are noted.

Hemolytic jaundice due to hemolysis of erythrocytes and, as a result, increased formation of bilirubin. An increase in the content of free bilirubin is one of the main signs of hemolytic jaundice.

In clinical practice, congenital and acquired functional hyperbilirubinemias are distinguished, caused by a violation of the elimination of bilirubin from the body (the presence of defects in enzymatic and other systems for the transfer of bilirubin through cell membranes and its glucuronidation in them). Gilbert's syndrome is a hereditary benign chronic disease that occurs with moderately severe non-hemolytic unconjugated hyperbilirubinemia. Posthepatitic hyperbilirubinemia Kalka - an acquired enzyme defect leading to an increase in the level of free bilirubin in the blood, congenital familial non-hemolytic Crigler-Najjar jaundice (absence of glucuronyl transferase in hepatocytes), jaundice in congenital hypothyroidism (thyroxine stimulates the enzyme glucuronyl transferase system), physiological neonatal jaundice, drug jaundice, etc. .

Disturbances in pigment metabolism can be caused by changes not only in the processes of heme breakdown, but also in the formation of its precursors - porphyrins (cyclic organic compounds based on the porphin ring, consisting of 4 pyrroles connected by methine bridges). Porphyrias are a group of hereditary diseases accompanied by a genetic deficiency in the activity of enzymes involved in the biosynthesis of heme, in which an increase in the content of porphyrins or their precursors is found in the body, which causes a number of clinical signs (excessive formation of metabolic products, causes the development of neurological symptoms and (or) an increase in skin photosensitivity).

The most widely used methods for the determination of bilirubin are based on its interaction with a diazoreagent (Ehrlich's reagent). The Jendrassik-Grof method has become widespread. In this method, a mixture of caffeine and sodium benzoate in acetate buffer is used as a "liberator" of bilirubin. The enzymatic determination of bilirubin is based on its oxidation by bilirubin oxidase. It is possible to determine unconjugated bilirubin by other methods of enzymatic oxidation.

Currently, the determination of bilirubin by the methods of "dry chemistry" is becoming more widespread, especially in express diagnostics.

Vitamins.

Vitamins are called irreplaceable low molecular weight substances that enter the body with food from the outside and are involved in the regulation of biochemical processes at the level of enzymes.

Similarities and differences between vitamins and hormones.

similarity- regulate metabolism in the human body through enzymes:

· vitamins are part of enzymes and are coenzymes or cofactors;

· Hormones or regulate the activity of already existing enzymes in the cell, or are inducers or repressors in the biosynthesis of the necessary enzymes.

Difference:

· vitamins- low molecular weight organic compounds, exogenous factors for the regulation of metabolism and come with food from the outside.

· Hormones- high-molecular organic compounds, endogenous factors synthesized in the endocrine glands of the body in response to changes in the external or internal environment of the human body, and also regulate metabolism.

Vitamins are classified into:

1. Fat soluble: A, D, E, K, A.

2. Water-soluble: group B, PP, H, C, THFA (tetrahydrofolic acid), pantothenic acid (B 3), P (rutin).

Vitamin A (retinol, antixerophthalmic) - the chemical structure is represented by a β-ionone ring and 2 isoprene residues; the need in the body is 2.5-30 mg per day.

The earliest and specific sign of hypovitaminosis A is hemeralopia (night blindness) - a violation of twilight vision. It occurs due to a lack of visual pigment - rhodopsin. Rhodopsin contains retinal (vitamin A aldehyde) as an active group - it is found in retinal rods. These cells (rods) perceive light signals of low intensity.

Rhodopsin = opsin (protein) + cis-retinal.

When rhodopsin is excited by light, cis-retinal, as a result of enzymatic rearrangements inside the molecule, passes into all-trans-retinal (in the light). This leads to a conformational rearrangement of the entire rhodopsin molecule. Rhodopsin dissociates into opsin and trans-retinal, which is a trigger that excites an impulse in the optic nerve endings, which is then transmitted to the brain.

In the dark, as a result of enzymatic reactions, trans-retinal is again converted into cis-retinal and, combining with opsin, forms rhodopsin.

Vitamin A also affects the growth and development of the integumentary epithelium. Therefore, with beriberi, damage to the skin, mucous membranes and eyes is observed, which manifests itself in pathological keratinization of the skin and mucous membranes. Patients develop xerophthalmia - dryness of the cornea of ​​\u200b\u200bthe eye, as there is a blockage of the lacrimal canal as a result of keratinization of the epithelium. Since the eye ceases to be washed with a tear, which has a bactericidal effect, conjunctivitis develops, ulceration and softening of the cornea - keratomalacia. With beriberi A, there may also be damage to the mucous membrane of the gastrointestinal tract, respiratory and genitourinary tract. Violated resistance of all tissues to infections. With the development of beriberi in childhood - growth retardation.

At present, the participation of vitamin A in the protection of cell membranes from oxidizing agents has been shown - that is, vitamin A has an antioxidant function.