RU2291440C1 - Method for diagnosing hepatitis transit into chronic stage - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves determining collagenase, collagenase inhibitor, protease inhibitor activity, free oxyproline and protein-bound oxyproline content. Collagenase activity increasing to 2.2±0.2 mcmole/l*h, protease inhibitor activity falling to 19.2±0.5 IU/ml and lower, collagenase inhibitor content falling to 2.1±0.2 IU/ml and lower, free oxyproline content growing to 1.2±0.02 IU/ml and protein-bound oxyproline content growing to 11.7±0.7 IU/ml, I stage of chronic transition is to be diagnosed. Collagenase activity increasing to 3.52±10.06 mcmole/l*h, collagenase inhibitor activity and protease inhibitor activity and free oxyproline content remaining the same, II stage of chronic transition is to be diagnosed. Collagenase activity increasing to 3.5±0.06 mcmole/l*h, protease inhibitor activity falling to 10.6±0.1 IU/ml and free oxyproline content growing to 1.524±0.06 IU/ml, III stage of chronic transition is to be diagnosed. Collagenase activity increasing to 4.06±0.02 mcmole/l*h, protease inhibitor activity falling to 9.0±0.28 IU/ml and lower, collagenase inhibitor content falling to 1.4±0.1 IU/ml, and protein-bound oxyproline content growing to 12.75±0.1 IU/ml, IV stage of chronic transition is to be diagnosed.

EFFECT: high accuracy of prognosis.

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Description

The invention relates to the field of medicine, hepatology and can be used to diagnose the stage of chronic hepatitis.

A known method for diagnosing the stage of chronicity of hepatitis by histological basis (V. Desmet et al, 1994), [11] in which determine the severity of fibrotic changes, and thereby evaluate the stage of chronicity of the process.

The known method is invasive - using a biopsy of the liver, a sampling of biological material is carried out and then histological examination is carried out with the calculation of the activity index (IGA) and histological index of fibrosis (GIF).)

However, the stage of chronicity of hepatitis reflects its temporary course and is characterized by the degree of liver fibrosis up to the development of cirrhosis. Cirrhosis is considered an irreversible stage of chronic hepatitis.

A new technical result is to increase the accuracy and information content while reducing the number of complications.

The problem is solved by a new method for diagnosing the stage of hepatitis chronicity in the liver, which consists in examining a patient, and blood serum is examined to determine the activity of collagenase, a collagenase inhibitor, a protease inhibitor, the content of free hydroxyproline and protein-bound hydroxyproline, and with an increase in collagenase activity to 2.2 ± 0, 2 μmol / l · h, a decrease in the activity of the protease inhibitor to 19.2 ± 0.5 IE / ml and lower, activity of a collagenase inhibitor to 2.1 ± 0.2 IE / ml and lower, an increase in the content of free oxyp olin to 1.2 ± 0.02 IE / ml, and protein-bound hydroxyproline to 11.7 ± 0.7 IE / ml diagnose stage I chronicity, with an increase in collagenase activity to 3.52 ± 0.06 μmol / l · h, an increase in the content of free oxyproline to 1.45 ± 0.02, with the same level of activity of the collagenase inhibitor and the activity of the protease inhibitor, the content of protein-bound oxyproline, stage II chronicity is diagnosed, with an increase in collagenase activity to 3.5 ± 0.06 μmol / l · h, a decrease in the activity of the protease inhibitor to 10.6 ± 0.1 IE / ml, an increase in the content of free oxyproline and up to 1.52 ± 0.01 IE / ml, stage III chronicity is diagnosed, with an increase in collagenase activity to 4.06 ± 0.02 μmol / l · h, a decrease in the activity of the protease inhibitor to 9.0 ± 0.28 IE / ml and below, and a collagenase inhibitor to 1.4 ± 0.1 IE / ml, an increase in the content of protein-bound oxyproline to 12.75 ± 0.1 IE / ml, stage IV chronicity is diagnosed.

The method is carried out in the following way.

The blood serum of the patient determines the activity of collagenase, a collagenase inhibitor, a protease inhibitor, the content of free oxyproline and protein-bound oxyproline and with an increase in collagenase activity to 2.2 ± 0.2 μmol / l · h, a decrease in the activity of the protease inhibitor to 19.2 ± 0, 5 IE / ml and below, a collagenase inhibitor to 2.1 ± 0.2 IE / ml and below, an increase in the content of free oxyproline to 1.2 ± 0.02 IE / ml, and protein-bound oxyproline to 11.7 ± 0.7 IE / ml diagnose stage I of chronicity, with an increase in collagenase activity to 3.52 ± 0.06 μmo b / l · h, an increase in the content of free oxyproline to 1.45 ± 0.02 was diagnosed as stage II of chronicity, with an increase in collagenase activity to 3.5 ± 0.06 μmol / l · h, a decrease in the activity of the protease inhibitor to 10.6 ± 0.1 IE / ml, an increase in the content of free oxyproline to 1.52 ± 0.01 IE / ml, stage III chronicity is diagnosed, with an increase in collagenase activity to 4.06 ± 0.02 μmol / l · h, a decrease in the activity of the protease inhibitor to 9.0 ± 0.28 IE / ml and lower, and the activity of a collagenase inhibitor to 1.4 ± 0.1 IE / ml, increasing the content of protein-bound hydroxyproline to 12 , 75 ± 0.1 IE / ml diagnose stage IV chronicity.

The proposed criteria are selected on the basis of an analysis of clinical trials of 60 patients with chronic hepatitis of various stages of chronicity of the process in the liver, who underwent histological studies of liver biopsy samples and determination of biochemical parameters in blood serum according to the proposed method.

So, in those patients where the serum collagenase activity was completely absent, the serum collagenase activity was not higher than 12.7 ± 0.04 μmol / l · h, the activity of the protease inhibitor was not lower than 26.4 ± 0.5 IE / ml, collagenase inhibitor was not lower than 3.86 ± 0.01 IE / ml, the content of free oxyproline was not higher than 0.6 ± 0.02 IE / ml, and protein bound was not higher than 6.7 ± 0.01 IE / ml.

In patients with stage I fibrosis, collagenase activity increased - 2.2 ± 0.2 μmol / l · h, the activity of the protease inhibitor decreased below 19.2 ± 0.5 IE / ml, the collagenase inhibitor decreased below 2.1 ± 0.2 IE / ml, the content of free oxyproline increased to 1.2 ± 0.02 IE / ml and protein-bound oxyproline to 11.7 ± 0.7 IE / ml.

In the second phase of fibrosis, collagenase activity increased to 3.52 ± 0.06 μmol / l · h, the content of free oxyproline increased to 1.45 ± 0.02. Other indicators did not change.

In stage III fibrosis, collagenase activity increased to 3.5 ± 0.06 μmol / l · h, the activity of the protease inhibitor decreased to 10.6 ± 0.1 IE / ml, the content of free oxyproline increased to 1.52 ± 0.01 IE / ml Other indicators did not change.

In stage IV of chronicity (cirrhosis), collagenase activity increased to 4.06 ± 0.02 μmol / l · h, the activity of the protease inhibitor decreased to 9.0 ± 0.28 IE / ml and lower, the activity of the collagenase inhibitor decreased to 1.4 ± 0.1 IE / ml, the content of protein-bound hydroxyproline increased to 12.75 ± 0.1 IE / ml. Other indicators remained the same.

Apparently, the revealed correlation is due to the following: biodegradation of collagen carried out by specific protease-collagenase [4, 6, 7] can be considered as activation of the collagenolysis system in response to increased collagen synthesis [3]. At the same time, the activity of the inhibitor link decreases — the activity of the protease inhibitor and collagenase inhibitor decreases [1]. Collagen degradation leads to an increase in the content of oxyproline and its fractions — free oxyproline and protein-bound oxyproline [10].

According to a puncture biopsy of the liver, which proves the presence of a certain stage of fibrosis, respectively, the stage of the process chronization and a parallel biochemical blood test for collagenase activity [8], collagenase inhibitors and a protease inhibitor [5], as well as determining the content of oxyproline - its fractions of free oxyproline and protein-bound oxyproline [2] it is possible to judge the existing shifts in the biochemical parameters of blood associated with an increase in the stage of fibrosis.

In stage I fibrosis, collagenase activity significantly increased (almost 2 times - 2.2 ± 0.04), collagenase inhibitor activity decreased (p <0.05) (almost 2 times - 2.0 ± 0.2) and The activity of the protease inhibitor decreased 1.5 times (19.2 ± 0.5), the content of free and protein bound oxyproline increased (p <0.05), respectively 1.2 ± 0.02 and 11.7 ± 0, 7 IE / ml.

In stage II fibrosis, collagenase activity increases (3 times - 3.2 ± 0.06), the content of free hydroxyproline (p <0.05) increases - more than 2 times - 1.45 ± 0.02 IE / ml.

In stage III fibrosis, collagenase activity increases (3.5 ± 0.06), the activity of the protease inhibitor decreases 10.6 ± 0.19 (almost 2 times in comparison with stage I), the content of free hydroxyproline increases 2 times - 1, 52 ± 0.01 (p <0.05) IE / ml.

Example 1. A 39-year-old patient, diagnosed with chronic hepatitis of viral etiology, type B, stage I fibrosis, moderate stage of activity, HBcor AgIgG +, HBsAG +, increased transaminases - 2 times, the liver was increased by 1.5 cm below the costal arch, painless, mild, positive PCR for HBV DNA. During a biochemical blood test, there were no violations of the protein-synthetic function of the liver, the prothrombin index was 90. A study was carried out according to the proposed method. During a biochemical blood test, the collagenase activity was 3.2 μmol / l · h, the protease inhibitor was 19.0 IE / ml, the activity of the collagenase inhibitor was reduced to 2.08, the content of free hydroxyproline to 1.25 IE / ml, protein-bound hydroxyproline up to 10.0 IE / ml. The data obtained made it possible to diagnose chronic hepatitis of the first stage of chronicity (stage I fibrosis).

Subsequent histological examination of the liver confirmed the diagnosis.

Example 2. Patient 32 years old, diagnosed with chronic hepatitis of viral etiology, type C, in the replication phase (positive PCR for HCV RNA), stage II fibrosis. During a biochemical blood test, transaminases were normal, albumin and prothrombin index were normal, collagenase activity increased to 3.58 μmol / l · h, free oxyproline content increased to 1.43 IE / ml. The diagnosis was made: chronic hepatitis, stage II chronicity.

With a histological examination of the liver, the diagnosis was confirmed.

Example 3. Patient, 36 years old, diagnosed with chronic hepatitis of viral etiology, type C, in the phase of replication of viral infection, minimal activity, stage III fibrosis. The patient did not show complaints, the liver was palpated 1 cm below the costal arch, painless, dense. Peripheral hepatic signs in the form of single telangiectasias were noted. During a biochemical blood test, transaminases were slightly increased, did not exceed more than 1.5 times, the albumin content was within the range of 38 g / l, the prothrombin index was 85. According to the data of an additional study, according to the proposed method, collagenase activity increased to 3.56 μmol / l · h, the protease inhibitor decreased to 11.6 IE / ml, the content of free oxyproline increased to 1.51 IE / ml: chronic stage III hepatitis was diagnosed with fibrosis, which was confirmed by paragraph ionic liver biopsy

Example 4. Patient C., 49 years old, diagnosed with cirrhosis of the liver of viral etiology, minimal stage of activity. During a biochemical study, transaminases were within normal limits, bilirubin concentration was 20 mg /%, of which direct - 12, indirect - 8, albumin - 35 g / l, prothrombin index - 82, an additional study was conducted according to the proposed method - collagenase activity increased to 4.04 μmol / l · h, the activity of the protease inhibitor decreased to 8.72 IE / ml, the collagenase inhibitor decreased to 1.5 IE / ml, the content of protein-bound hydroxyproline increased to 12.74 IE / ml, the diagnosis of stage IV fibrosis liver cirrhosis was confirmed by paragraph ionic liver biopsy.

Thus, using the proposed method, it is possible to diagnose with high accuracy and informativeness the stage of chronicity of the process in the liver in patients with hepatitis, avoiding the use of the invasive method associated with a liver biopsy, which is contraindicated at some stages of the disease, as it can cause bleeding.

Literature

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2. Kuznetsova TP, Proshina L.Ya., Privalenko L.I. Modification of the determination of serum oxyproline // Laboratory work. - 1982. - No. 8. - S.8-10.

3. Logvinov A.S., Block Yu.E. Chronic hepatitis and cirrhosis of the liver. - M .: Medicine, 1987.

4. Maskevichus Z.K. Mechanisms and the role of collagen biodegradation in pathology / Z.K. Maskevichus // Archive of Pathology. - 1987. - T. 49, No. 6. - C.3-9.

5. Nartikova V.F., Paschina T.S. Unified method for determining the activity of α ₁ antitrypsin and α ₂ macroglobulin in human serum (plasma) // Problems of medical chemistry. - 1979. - T. 25, No. 4. - S. 494-499.

6. Serov VV Connective tissue / V.V. Serov, A.B.Shekhter. - M .: Nauka, 1981. - 456 p.

7. Slutsky L.I. Biochemistry of normal and pathological changes in connective tissue / L.I. Slutsky. - M .: Nauka, 1969 .-- 345 p.

8. Sharaev P.N., Pishkov V.N., Zvorygina N.G. Determination of collagenolytic activity of blood plasma // Laboratory. - 1987. - No. 1, - S.60-62.

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Table 1
The stages of chronic hepatitis disease by histological signs - the degree of fibrosis (histological index (GI fibrosis)) GI Degree of fibrosis Morphological features 0 absent absent one weak fibrosis and expansion of portal tracts 2 moderate periportal fibrosis 3 heavy portocentral septa four cirrhosis cirrhosis

Claims (1)

A method for diagnosing the stage of chronic hepatitis in the liver, which consists in examining a patient, characterized in that blood serum is examined to determine the activity of collagenase, a collagenase inhibitor, a protease inhibitor, the content of free hydroxyproline and protein-bound hydroxyproline, and with an increase in collagenase activity to (2.2 ± 0, 2) μmol / l · h, a decrease in the activity of the protease inhibitor to (19.2 ± 0.5) IE / ml and below, activity of a collagenase inhibitor to (2.1 ± 0.2) IE / ml and below, an increase in the content of free oxyproline to (1.2 ± 0.02) IE / ml, and protein-bound oxyproline to (11.7 ± 0.7) IE / ml diagnose stage I of chronicity, with an increase in collagenase activity to (3.52 ± 0.06) μmol / l · h, an increase in the content of free oxyproline to ( 1.45 ± 0.02) IE / ml with the same level of collagenase inhibitor activity and protease inhibitor activity, protein-bound hydroxyproline content, stage II chronicity is diagnosed, with an increase in collagenase activity to (3.5 ± 0.06) μmol / l · h, a decrease in the activity of the protease inhibitor to (10.6 ± 0.1) IE / ml, an increase in the content of free oxyproline to (1.5 2 ± 0.01) IE / ml diagnose stage III of chronicity, with an increase in collagenase activity to (4.06 ± 0.02) μmol / l · h, a decrease in the activity of the protease inhibitor to (9.0 ± 0.28) IE / ml and below, and a collagenase inhibitor to (1.4 ± 0.1) IE / ml, an increase in the content of protein-bound oxyproline to (12.75 ± 0.1) IE / ml, stage IV chronicity is diagnosed.