RU2459209C1 - Method for prediction of acne severity level in patients with intestinal dysbiosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method for prediction of acne severity level in patients with intestinal dysbiosis consists in the fact that bacterial feces analysis aims at the qualitative and quantitative composition, and biochemical feces analysis - for short-chain fatty acids. If observing decreasing the content of bifidus bacteria to 10⁷ CFU/g and less, lactic acid bacilli to 10⁵ CFU/g and less, typical Escherichia to 10⁵ CFU/g, increasing the content of hemolytic Escherichia to the concentration of 10⁵-10⁷ CFU/g, as well as the content of acetic acid more than 0.634 mg/g, of propionic acid less than 0.189 mg/g, of butyric acid less than 0.176 mg/g, the potential aggravation of intestinal dysbiosis worsening the clinical course of acne is predicted.

EFFECT: use of the invention provides earlier diagnosing and prediction of the acne transition in the course of conventional traditional therapy into a more severe clinical version.

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Description

The invention relates to medicine, namely to dermatovenerology, and can be used for a laboratory method for predicting the course of severity of acne in patients with intestinal dysbiosis.

The role of changes in the composition and activity of microbiocenoses in the pathogenesis of a wide range of human diseases is currently under active discussion. Normal microflora is the target of the negative impact of various in nature factors. Regardless of the cause that caused them, changes in microbial ecology lead to a violation of the protective, metabolic, regulatory properties of microbiota, which inevitably affects all processes directly or indirectly associated with the functioning of human microbiocenoses. Intestinal dysbiosis is a clinical and microbiological syndrome characterized by quantitative, qualitative and topographic changes in the normal intestinal microflora, associated with the onset, development and clinical course of most diseases of the human internal organs. Numerous experimental and clinical data indicate the involvement of dysbiotic disorders of the intestinal microflora in the pathogenesis of a number of diseases at different stages. These include skin diseases, including acne. Intestinal dysbiosis is characterized by a certain fraction of treachery, which consists in the fact that for a long time there may not be any obvious clinical signs. In the future, under conditions of dysbiosis, the risk of development increases and the course of various diseases (gastrointestinal, cardiovascular, allergic, infectious, skin, etc.) is aggravated. It must be remembered that the fact that dysbiotic changes in the intestine are manifested by a worsening of the course of the underlying disease is clinically significant. At the same time, with the increase in the severity of dysbiosis, the severity of clinical symptoms increases, their life span is lengthened, the general condition is aggravated, treatment outcome indicators worsen, relapses and the complicated course of the underlying disease are often noted, and the quality of life of patients worsens. A clear relationship between dysbiotic disorders in the intestine and manifestations of skin diseases was revealed.

In practice, for the diagnosis of intestinal dysbiosis, 15-20 types of microorganisms contained in the feces are studied, in particular the number of bifidobacteria, lactobacilli, enterobacteria, E. coli, protea, enterococcus, Staphylococcus aureus, Pseudomonas aeruginosa and Candida genus fungi. The severity of dysbiosis is assessed by the degree of decrease in the number of bifidobacteria and other obligate microorganisms, as well as an increase in the number of conditionally pathogenic microorganisms (UPM).

For an objective assessment of the composition of intestinal microflora, a microbiological study of feces is used, which is an integral indicator of the state of intestinal microbiocenosis. Recently, the method of volatile fatty acids in various biological substrates has become widespread, which, in our opinion, has great prospects in assessing local and systemic manifestations of microecological disturbances, which has high resolution, cheap research costs, quick response, and, possibly, test-control of the effectiveness of the therapy.

The modern classification of dysbiosis is based on a combination of clinical and microbiological research data. Four degrees of severity of intestinal dysbiosis are distinguished [I. B. Kuvaeva, K. S. Ladodo, 1991].

I severity is characterized by a decrease of 1-2 orders of magnitude of bifidobacteria and lactobacilli, E. coli. Severity II - against a background of a decrease in the content of bifidobacteria and lactobacilli by 3-4 orders of magnitude, an increase in the number of conditionally pathogenic microorganisms of staphylococci, protea is detected. Conditionally pathogenic flora acquires hemolytic properties, that is, it is able to be aggressive. Grade III severity is characterized by a significant decrease in the number of anaerobes (bifidobacteria and lactobacilli to 10 ⁵ -10 ⁶ ) and a gradual increase in aerobes. Conditionally pathogenic flora acquires pronounced aggressive properties. At this stage, fungi of the genus Candida, Proteus, Klebsiella, enterobacteria, staphylococci are found in large numbers. IV severity is characterized by the absence of bifidobacteria, a significant decrease in the number of lactobacilli and E. coli. Significant changes in the quantitative ratios of obligate and facultative microorganisms, their biological properties, and the accumulation of entero- and cytotoxins are noted. The functions of the intestinal microflora and, first of all, the antagonistic ones are sharply inhibited, which leads to a pronounced activation of the conditionally pathogenic flora, severe digestive disorders, destructive changes in the mucous membrane of the digestive tract, and a decrease in nonspecific resistance.

Diagnosing acne is usually not difficult. Acne is characterized by open and closed comedones (black and white eels), which can be single or usually multiple with erythematous papules and pustules, often localized on the face and upper body. Various classifications are used to determine the severity of the disease. The following classification of the disease was proposed at the 20th World Congress in Paris in 2002 [Samtsov AV, 2009]: mild - comedones, papulopustules (up to 10 elements); the middle degree - papulopustules (from 10-25 elements), nodes (up to 5 nodes); severe degree - papulopustules (from 26-50 elements), nodes (more than 5).

The prototype of the invention is a method for differential diagnosis of the severity of acne, which consists in the fact that the activity of myeloperoxidase and amylase is determined in the patient’s saliva with a myeloperoxidase activity of 35.23-42.30 μm mol / mg, and amylase 34.88-42.88 mg / (g × ml) diagnose the first or second severity of acne, and with a myeloperoxidase activity of more than 42.30 μmol / mg, and amylase more than 42.88 mg / (g × ml) diagnose a third or fourth degree of severity [patent RU 2323442, 2008]. However, this method does not consider the severity of the course of acne on the background of intestinal dysbiosis.

The objective of the invention is the early diagnosis and prediction of the severity of acne in patients with intestinal dysbiosis.

EFFECT: obtaining criteria for predicting the severity of acne in patients with intestinal dysbiosis.

The specified technical result is achieved by the fact that in the method for predicting the severity of acne, including a biochemical study of a biosubstrate, according to the invention before treatment and on the 14th day after treatment, the concentration of short-chain fatty acids in the feces of patients with intestinal dysbiosis is determined as a biochemical study of biosubstrate, additionally bacteriological examination of feces and with the value of the content of acetic acid above 0.634 mg / g, propionic acid below 0.189 mg / g, butyric acid below 0 , 176 mg / g, as well as reducing the content of bifidobacteria to 10 ⁷ CFU / g or less, lactobacilli to 10 ⁵ CFU / g or less, typical Escherichia to 10 ⁵ CFU / g, increasing the content of hemolytic Escherichia to a concentration of 10 ⁵ -10 ⁷ CFU / g predict aggravation of intestinal dysbiosis and the transition of acne to a more severe clinical variant.

The proposed method for predicting the severity of acne in patients with intestinal dysbiosis is as follows: the qualitative and quantitative composition of feces is determined by the bacteriological method, and short-chain fatty acids (CFA) (C2-C6) in biosubstrate (feces) are determined by the method of volatile fatty acids. The study is carried out before treatment and on the 14th day of treatment. With a decrease in the content of bifidobacteria to 10 ⁷ CFU / g or less, lactobacilli to 10 ⁵ CFU / g or less, typical Escherichia to 10 ⁵ CFU / g, an increase in the content of hemolytic Escherichia to a concentration of 10 ⁵ -10 ⁷ CFU / g, and also if indicators of acetic acid above 0.634 mg / g, propionic acid below 0.189 mg / g, butyric acid below 0.176 mg / g predict the possibility of worsening intestinal dysbiosis, worsening the severity of acne. Moreover, in patients with acne during the course of traditional therapy, the disease can transfer to a more severe clinical variant.

According to the research protocol, the clinical dynamics of the process on the skin, as well as laboratory testing of 35 observed acne patients with intestinal dysbiosis were performed before treatment and on the 14th day of traditional drug therapy.

The method of bacteriological examination of feces for dysbiosis included the following stages: sampling, transportation and preparation for sowing the test material; study of qualitative and quantitative characteristics of the microflora of feces; registration of the results, the conclusion of the bacteriologist on the state of the intestinal microflora.

The method for determining FFA in a biosubstrate consists of two stages: the process of sample preparation and directly gas-liquid chromatographic analysis (GLC). The method was registered in the Ministry of Health and Social Development of the Russian Federation (registration number FS-2006/030-u dated 03/17/2006)

With a decrease in the content of bifidobacteria to 10 ⁷ CFU / g or less, lactobacilli to 10 ⁵ CFU / g or less, typical Escherichia to 10 ⁵ CFU / g, an increase in the content of hemolytic Escherichia or other opportunistic bacteria to a concentration of 10 ⁵ -10 ⁷ CFU / g, and also if acetic acid is higher than 0.634 mg / g, propionic acid is lower than 0.189 mg / g, butyric acid is lower than 0.176 mg / g, the possibility of aggravation of intestinal dysbiosis worsening the course of acne is predicted. Moreover, in patients with acne during the course of traditional therapy, the disease can transfer to a more severe clinical variant.

These indicators are components of bacteriological and biochemical studies of feces, are available for implementation in the clinical diagnostic laboratory.

In a number of patients with acne and concomitant intestinal dysbiosis, in spite of the traditional therapy, an aggravation of the process on the skin was observed, which was manifested by the transition of the papule to the pustule, pustules to the node. Along with the worsening clinical picture, marked changes were noted in bacteriological and biochemical studies of feces.

In the available scientific, medical and patent literature, no information was found on bacteriological and biochemical studies of feces in order to predict the transition of acne to a more severe form. Thus, the claimed invention meets the patentability criterion of "novelty."

The studies of the authors for the first time proved the possibility of accurate prediction of the transition of acne into a more severe clinical variant based on bacteriological and biochemical analysis of feces. Thus, the claimed invention meets the criterion of "inventive step".

The proposed method evaluated the likelihood of a transition of acne to a more severe degree in 35 patients and in all cases the specified technical result was achieved. The method is easily reproducible in a diagnostic laboratory. Thus, the claimed invention meets the criterion of "industrial applicability".

The proposed forecasting method is illustrated by the following clinical examples (see table 1, 2):

Example 1. Patient V., 17 years old, complained of common rashes on the face and neck. The duration of the rash is 4 years. As well as a decrease in appetite, constipation, fatigue, headache. He received treatment several times: massage with liquid nitrogen, facial cleansing, lotion with chloramphenicol, ointment was skinned, the effect of the treatment was not received.

Status localis: multiple small papulopustular elements (24) and nodes (in the amount of 3 pieces), mainly in the cheeks and margins of the lower jaw. Pronounced greasy shine.

Bacteriological examination of feces was carried out: bifidobacteria 10 ⁷ CFU / g, lactobacilli 10 ⁵ CFU / g, E. coli typical 10 ⁶ CFU / g, hemolytic E. coli 10 ⁴ CFU / g. A biochemical study of feces revealed: indicators of acetic acid 0.652 mg / g, indicators of propionic acid 0.179 mg / g, butyric acid 0.169 mg / g.

Diagnosed with papulopustular form of acne, moderate.

Concomitant diagnosis: intestinal dysbiosis, II severity.

The patient was treated according to standard therapy, which included antibiotic therapy, vitamin therapy, and topical treatment.

On the 14th day of treatment, despite the ongoing therapy, an increase in the clinical process was observed, which was visually manifested by an increase in the number of elements. There were 32 papulopustular elements and the number of nodes increased to 7 pieces.

The indicators of fecal bacteriological analysis on the 14th day were: bifidobacteria 10 ⁶ CFU / g, lactobacilli 10 ⁵ CFU / g, E. coli typical 10 ⁵ CFU / g, hemolytic E. coli 10 ⁶ CFU / g. A biochemical study of feces revealed: indicators of acetic acid 0.674 mg / g, indicators of propionic acid 0.172 mg / g, butyric acid 0.154 mg / g.

On the 14th day, treatment with a prebiotic drug was added to correct dysbiotic disorders. One month after this treatment, the stool bacteriological analysis indicators were: bifidobacteria 10 ⁸ CFU / g, lactobacilli 10 ⁹ CFU / g, E. coli typical 10 ⁷ CFU / g. A biochemical study of feces revealed: indicators of acetic acid 0.632 mg / g, indicators of propionic acid 0.183 mg / g, butyric acid 0.173 mg / g.

Correction of the therapy given the concomitant pathology made it possible to achieve a regression of the process, to prevent the development of a more severe clinical variant.

Example 2. Patient G., 18 years old, complained of rashes on the face. The disease is 5 years old. Rashes are constantly progressing. The heredity of acne is burdened by the father. The patient received external treatment - lotions with sulfur, Zinerit solution. Status localis: multiple closed comedones, papulopustular elements (10 elements) in the face area (mainly in the corners of the lower jaw). Pores on the face are expanded in the area of the T-zone. Pronounced greasy shine.

Preliminary diagnosis: papulopustular form of acne, mild.

Bacteriological examination of feces was carried out: bifidobacteria 10 ⁷ CFU / g, lactobacilli 10 ⁵ CFU / g, E. coli typical 10 ⁶ CFU / g. A biochemical study of feces revealed: indicators of acetic acid 0.647 mg / g, indicators of propionic acid 0.175 mg / g, butyric acid 0.172 mg / g.

Diagnosed with papulopustular form of acne, mild severity.

Concomitant diagnosis: intestinal dysbiosis, II severity.

The patient was treated according to standard therapy, which included only vitamin therapy and local treatment.

On the 14th day of treatment, despite the ongoing therapy, an increase in the clinical process was observed, which was visually manifested by an increase in the number of elements. There were 20 papulopustular elements and 2 nodes appeared.

The indicators of fecal bacteriological analysis on the 14th day were: bifidobacteria 10 ⁶ CFU / g, lactobacilli 10 ⁵ CFU / g, E. coli typical 10 ⁵ CFU / g, hemolytic E. coli 10 ⁵ CFU / g. A biochemical study of feces revealed: indicators of acetic acid 0.659 mg / g, indicators of propionic acid 0.181 mg / g, butyric acid 0.161 mg / g.

On the 14th day, treatment with a prebiotic drug was also added to correct dysbiotic disorders. One month after the treatment of fecal indicators bacteriological analysis made bifidobacteria September ¹⁰ CFU / g lactobacilli September ¹⁰ CFU / g, E. coli typical 10 ⁷ cfu / g. A biochemical study of feces revealed: indicators of acetic acid 0.633 mg / g, indicators of propionic acid 0.187 mg / g, butyric acid 0.175 mg / g.

Table 1 Bacteriological and biochemical indicators of feces in patient B. (according to example No. 1) in dynamics indicators before treatment on the 14th day of treatment after treatment Bifidobacteria, CFU / g 10 ⁷ 10 ⁶ 10 ⁸ Lactobacilli, CFU / g 10 ⁵ 10 ⁵ 10 ⁹ Typical Escherichia, CFU / g 10 ⁶ 10 ⁵ 10 ⁷ Hemolytic Escherichia, CFU / g 10 ⁴ 10 ⁶ - Acetic acid, mg / g 0.652 0.674 0.632 Propionic acid, mg / g 0.179 0.172 0.183 Butyric acid, mg / g 0.169 0.154 0.173

table 2 Bacteriological and biochemical indicators of feces in patient G. (according to example No. 2) in dynamics indicators before treatment on the 14th day of treatment after treatment Bifidobacteria, CFU / g 10 ⁷ 10 ⁶ 10 ⁹ Lactobacilli, CFU / g 10 ⁵ 10 ⁵ 10 ⁹ Typical Escherichia, CFU / g 10 ⁶ 10 ⁵ 10 ⁷ Hemolytic Escherichia, CFU / g - 10 ⁵ - Acetic acid, mg / g 0.647 0.659 0.633 Propionic acid, mg / g 0.175 0.181 0.187 Butyric acid, mg / g 0.172 0.161 0.175

Claims (1)

A method for predicting the severity of acne, including a biochemical study of biosubstrate, characterized in that before treatment and on the 14th day after treatment, as a biochemical study of biosubstrate, the concentration of short-chain fatty acids in the feces of patients with intestinal dysbiosis is determined, an additional bacteriological study of feces is carried out and if the content of acetic acid is higher than 0.634 mg / g, propionic acid is lower than 0.189 mg / g, butyric acid is lower than 0.176 mg / g, and also the content of bifidobacteria is reduced to 10 ⁷ CFU / g or less, lactobacilli up to 10 ⁵ CFU / g or less, typical Escherichia up to 10 ⁵ CFU / g, increased hemolytic Escherichia to a concentration of 10 ⁵ -10 ⁷ CFU / g predict aggravation of intestinal dysbiosis and the transition of acne to a more severe clinical option.