RU2442165C1 - Method of forecasting increasing of individual risk of chronic obstructive pulmonary disease associated with coronary heart disease - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention in the field of medicine refers to a method of forecasting increasing of individual risk of chronic obstructive pulmonary disease (copd) associated with coronary heart disease (chd). The essence of the method is in calculation of smoker index and number of pack-years, determine the degree of nicotine-associated risk and, accordingly, the intensity of smoking. Then analysis of polymorphousgene locus GSTM 1 and GSTT 1 is carried out by PCR method. Individuals-carriers of GSTs homozygous deletion with high nicotine-associated risk of chronic obstructive pulmonary disease associated with coronary heart disease.

EFFECT: using the method allow to predict the increasing of individual risk of COPD associated with coronary artery disease, prior to the development of a pathological process or in the early stages of the disease.

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Description

The invention relates to the field of predictive (predictive) medicine, and in particular to methods for predicting the risk of various diseases of the human respiratory and cardiovascular systems, in particular, to identifying the risk of developing chronic obstructive pulmonary disease (COPD) associated with coronary heart disease (CHD). The invention can be used in the field of predictive (preventive), diagnostic and preventive medicine.

COPD is one of the leading causes of adult morbidity and mortality worldwide and causes significant economic and social damage [1]. This lung pathology is widespread among people of working age throughout the world, including people from 30 years and older. IHD is the most common disease of the cardiovascular system. The proportion of IHD among the causes of death from cardiovascular disease (CVD) is 43-88% and increases sharply in people over 45 years of age [2]. This is one of the reasons for the temporary and permanent disability of the population in the developed countries of the world. According to large population studies, the risk of death from CVD in patients with COPD is 2-3 times higher and makes up about 50% of the total number of deaths. Given the commonality of risk factors, the presence of a comorbid pathology in the form of COPD associated with IHD is more often the rule than the exception [3]. The compatibility of IHD and COPD according to various studies in older age groups reaches more than 60%, and the 15-year survival of such patients is no more than 25% [2]. Dysfunction of the external respiration of patients with COPD with concomitant coronary heart disease is characterized by a more severe nature of respiratory disorders than in patients with “pure” COPD and is accompanied by an earlier development of emphysema [4]. Thus, in connection with the high prevalence of the combination of IHD and COPD, especially in the elderly, and the proven interdependent effect of these diseases, deeper studies are needed in this area in order to develop effective and safe treatment and prevention of the progression of both pathological processes.

Currently, the diagnosis of COPD and coronary heart disease is carried out by means of relatively long clinical observations with appropriate tests [5]. Diagnosis of COPD includes collecting an anamnesis, taking into account clinical manifestations, laboratory and instrumental methods of research. Sputum cytology, a clinical blood test, chest x-ray, ventilation and gas exchange function analysis of the lungs are part of the necessary diagnostic program for examining COPD patients. The leading role in the diagnosis of COPD is the study of the function of external respiration, since one of the main and mandatory signs of COPD is the presence and progression of bronchial obstruction. Spirometry is the gold standard in the diagnosis of COPD, as its indicators are best standardized, reproduced and are the most objective. According to the Global Strategy for the Diagnosis, Treatment, and Prevention of Chronic Obstructive Pulmonary Disease [1], currently in clinical practice the “gold standard” for diagnosing the processes of irreversible and reversible obstruction of the bronchi is a method of functional investigation of the function of external respiration (VFD) with a post-bronchodilation test, in particular estimation of forced expiratory volume in 1 sec (FEV ₁ ) [6]. In accordance with this, OFB _{1 is} considered a kind of “gold standard” for the diagnosis of COPD in screening studies. The disadvantage of this method is that diagnostically significant changes in FEV ₁ are detected much later, when pathomorphological processes in the bronchi that underlie remodeling have already been launched, there are also a variety of reasons for the decrease in FEV ₁ (except for COPD), in particular, a decrease in FEV ₁ may be due to extrapulmonary reasons: violation of breathing regulation, increased chest resistance, respiratory muscles weakness, etc. Various methods can be used to diagnose coronary heart disease: ECG recording at rest, holt ECG monitoring, echocardiography, various stress tests (treadmill test and bicycle ergometry), pharmacological tests, stress echocardiography, radioisotope methods, serial computed tomograms, magnetic resonance imaging with contrasting coronary arteries, coronary angiography. Currently, clinical protocols for diagnosis and treatment separately for COPD and IHD have been introduced into clinical practice. However, standards for the diagnosis and treatment of concomitant pathology have not been developed. Therefore, the doctor in clinical practice has to rely on the diagnostic criteria of an isolated pathology, although the current level of diagnosis has shown the real possibility of combining COPD and IHD. In addition, society has serious social prerequisites for the simultaneous development of these diseases: the widespread prevalence of smoking, air pollution with various pollutants, stress factors, lack of physical activity, and unhealthy diet. The course of the combined pathology is characterized by certain clinical features and an unfavorable prognosis. Therefore, the main task of the doctor at the stage of diagnosis is the early detection of the diagnostic criteria of IHD in a patient with COPD. Diagnosis should be focused and predictive, carried out using additional research methods.

In the onset and progression of COPD and IHD, a large role is played by a genetic predisposition and general risk factors, the main of which is smoking [3]. The key links in these diseases are lipid and protein peroxidation, activation of cytokine mechanisms (increase in C-protein, interleukins, tumor necrosis factor-alpha) with the involvement of the immune, endocrine systems with the release of hormones and neurotransmitters into the bloodstream, which is associated with increased consumption volatile plastic biological substrates in the mucous membranes of the bronchi and vascular endothelium [7-11]. Against the background of oxidative stress, the adrenal glands are stimulated with the release of adrenaline into the blood, leading to vasoconstriction, an increase in the volume of circulating blood, general peripheral vascular resistance and an increase in blood pressure, leading to the development of IHD. Against the background of a long-term pathological process (inflammation) in the lung, as a result of which the bronchial tree is rearranged and emphysema develops, hypertension of the pulmonary circulation with right ventricular myocardial hypertrophy, the so-called "pulmonary heart," is gradually forming. Respiratory hypoxemia, an increase in pre- and afterload (the basis of IHD) also contribute to hypertrophy and the formation of connective tissue cardiosclerosis of the left ventricle with an increase in the course of COPD and IHD [7, 9, 11, 12].

The problem of predictive diagnosis of COPD and coronary heart disease is extremely relevant, as evidenced by the following facts. COPD and coronary heart disease refers to common diseases, and there is a clear tendency to increase. According to the European Respiratory Society, only 25% of cases are diagnosed in a timely manner. In the United States, the incidence of COPD is approaching 15 million people and is one of the most common diseases in which mortality continues to increase. According to official figures, in Russia there are about 1 million patients with COPD, but in reality their number is an order of magnitude higher. In addition, the insidiousness of COPD lies in slow, but steady progression, severe clinical symptoms appear only in the advanced stage of the disease. In the early stages of COPD, it proceeds covertly without constant clinical symptoms. An urgent problem of COPD is its predictive diagnosis for the purpose of timely prevention aimed at key links in the pathogenesis of COPD. Unfortunately, predictive diagnosis and diagnosis of the early stages of this pathology in our country is practically absent, because There are no specific genetic or biochemical markers for determining the early stages of this disease.

Earlier, some researchers made attempts to develop methods for predicting COPD using laboratory [13, 14], instrumental [15, 16] and laboratory-instrumental methods [17]. A known method for predicting the progression of COPD based on determining the level of secretion of transforming growth factor β1 (TGF-β1) and fibroblast growth factor (bFGF) [13]. To assess the risk of developing COPD in long-term smokers, a method for predicting the disease was proposed based on determining the level of activity of α ₁ antitrypsin and α ₂ macroglobulin in sputum [14]. A method for predicting a stable course of COPD was developed on the basis of taking into account the results of measurement of FEV ₁ in l, changes in forced expiratory volume in the first second in% of the initial value after a bronchial provocative pharmacological test with a 0.1% solution of acetylcholine chloride [15]. A method for predicting COPD was also proposed based on the measurement of densitometry and planimetry in% of both lungs, determining the FEV value of ₁ in% to the proper [16]. A known method for predicting bronchial obstruction in chronic bronchitis, which consists in using quantitative laboratory criteria for the risk of developing bronchial obstruction, including determining the level of IL-1β in blood serum, saliva and bronchoalveolar fluid [17].

There are also methods for predicting the course of coronary heart disease based on the determination of total cholesterol [18] and serum lipids [19]. Known methods for predicting the course of coronary heart disease and the severity of chronic heart failure in patients with coronary artery disease based on the determination of titers of herpes simplex virus antigen and antiherpetic antibodies and determination of the concentration of IgG to herpes simplex virus type 1 in the patient’s blood serum [20, 21]. There are methods for predicting the risk of developing coronary heart disease based on determining the type of constitution and somatic signs of sex hormone imbalance by calculating the index of somatic sexual differentiation (in women of childbearing age) or only using an instrumental method of research (determining the vasomotor function of the endothelium) [22, 23]. There is also a method for predicting coronary heart disease in miners with chronic dust bronchitis, taking into account the following indicators (blood group of the ABO system, growth, constitutional-morphological type, the presence of a diagonal fold of the earlobe, the presence of arterial hypertension, smoking, IHD burdened) [24].

All of the above methods have significant drawbacks: they are laborious, time consuming, limited in the use of biological material, do not allow to determine the predisposition to the development of the pathological process and in the early stages of the disease. For some of them, there is a need for multiple studies, because results change throughout life. These methods also have one significant drawback - they do not allow to predict the individual risk of developing COPD associated with coronary heart disease. In addition, among the already known forecasting methods, the authors of the present invention were unable to find methods for predicting COPD and IHD based on an assessment of the degree of nicotine-associated risk in combination with molecular genetic indicators.

It is known that one of the leading modifiable risk factors for developing COPD and IHD is smoking [3]. The individualization of the body's response to tobacco combustion products can be determined by the polymorphism of xenobiotic detoxification genes. Glutathione-mediated detoxification is key in ensuring cell resistance to lipid peroxidation, free radicals, protein alkylation, and the prevention of DNA breakdowns [25-28]. Glutathione transferase genes (GSTs), in particular GSTM1 and GSTT1, play an important role in the biotransformation of the chemicals that make up cigarettes [27, 28]. In the studied available literature sources, the authors did not find mention of a method for predicting the individual risk of developing COPD associated with IHD. But in the literature there is information about the change in GSTs in COPD. In previous studies by foreign and domestic authors, groups of COPD patients were studied, but without association with IHD [1, 3, 25-28]. In articles [29–31], the GSTM1 and GSTT1 genes were studied, which play an important role in the biotransformation of the chemicals that make up cigarettes and are of the greatest interest in revealing the mechanism of COPD development. It has been suggested that homozygous deletions of the GSTT1 gene are not specific predisposing factors, but may be related to non-cancer lung diseases [31]. It was shown in [25, 26] that a homozygous deletion (zero genotype) of GSTM1 does not always have an independent value, but in combination with deletions of other genes (including glutathione transferases) significantly increases the risk of developing a pathological condition and aggravates its course . The results obtained by Sram RC suggest a greater exposure of individuals carrying homozygous GSTM1 deletion to the risk of developing COPD [32]. When conducting a cohort study of SAPALDIA on the association of glutathione transferase genes (GSTT1 and GSTM1) in patients with COPD, a statistically significant relationship was found between an isolated homozygous deletion of GSTT1 (or in combination with a homozygous deletion of GSTM1) and a decrease in FEV ₁ in men regardless of smoking status [25 ]. Studies have shown a relationship between glutathione transferase polymorphisms with a risk of developing COPD, decreased pulmonary function, and a genotoxic effect in tobacco smoking [26, 30, 33].

The basis of the present invention is to create a method for predicting an individual risk of developing COPD associated with coronary heart disease.

Given that smoking is the most important risk factor, the authors investigated homozygous deletions of two glutathione transferase genes GSTM1 and GSTT1, which provide detoxification of tobacco combustion products. It is known that to determine the nicotine-associated risk, indicators such as the smoking person index (ICH) and the number of pack-years (PLP) are used [1]. Therefore, in order to identify risk groups and provide adequate prevention of COPD and IHD, the authors of the present invention studied molecular genetic markers GSTM1 and GSTT1 in combination with indicators of ICH and EML in groups of patients with early stages of COPD and COPD associated with stable angina pectoris and in healthy individuals . Analysis of polymorphic loci of these genes was performed using the PCR method. A high incidence of homozygous deletions of GSTM1 and GSTT1 was established in patients with COPD and COPD associated with IHD, and the zero genotype (homozygous deletion) of GSTT1 was significantly associated only with the presence and severity of COPD associated with stable angina pectoris (p less than 0.05). In the presence of a combined null genotype of glutathione transferases GSTM1 and GSTT1 (namely, a homozygous deletion for both genes), the relative risk of a combined pathology - COPD associated with IHD (p less than 0.05) is increased. When comparing the frequencies of homozygous deletions of the GSTT1 and GSTM1 genes in patients with stage I-II COPD, no significant differences were found. Therefore, the identification of molecular genetic markers may allow their use to highlight risk groups and provide adequate prevention of COPD associated with IHD.

The inventive method consists in the fact that firstly, such indicators of nicotine-associated risk as the smoking person index (UIC) and the number of pack-years (NPI) are calculated, then biological material is collected, then DNA is extracted, the zero genotypes GSTM1 and GSTT1 are detected by PCR followed by gel electrophoresis. DNA amplification is carried out using reagent kits for the detection of polymorphic markers GSTM1 and GSTT1. Amplification products are analyzed by electrophoresis and visualized using a transilluminating gel-documenting system and in individuals carrying homozygous deletions of GSTs with a high degree of nicotine-associated risk (with an ICF of more than 120 and an ECL of more than 20 pack-years) they predict an increase in the relative risk of a combined pathology - COPD associated with IHD.

In this case: a) the presence of homozygous deletions of GSTT1 in combination with a high degree of nicotine-associated risk indicates a relative risk of a combined pathology - COPD associated with IHD; b) the presence of homozygous deletions of GSTT1 in combination with a high degree of nicotine-associated risk also indicates a high relative risk of the occurrence of a combined pathology - COPD associated with IHD; c) the presence of homozygous deletions of GSTM1 and GSTT1 in combination with a high degree of nicotine-associated risk indicates a high relative risk of a combined pathology - COPD associated with IHD; d) the presence of homozygous deletions of GSTM1 and GSTT1 in combination with a high degree of nicotine-associated risk indicates a very high relative risk of a combined pathology - COPD associated with IHD.

The research process included the following steps.

1) Determination of ICH, EPL to assess the intensity of smoking according to the formulas:

TIC = number of cigarettes smoked per day * number of months per year during which a person smokes. TIC more than 120 - a malicious smoker.

CPL = number of cigarettes smoked per day / 20 * smoking experience (years).

CPL more than 10 pack-years - permanent; CPL more than 20 pack-years - malicious smoker.

2) Blood sampling.

3) Isolation of nucleic acids. DNA is isolated from peripheral blood leukocytes using phenol-chloroform extraction.

4) PCR or amplification itself. The DNA isolated in the first stage is added to the solution containing the mixture of nucleotides, PCR buffer, polymerase and primers (sequences of oligonucleotide primers and methods for identifying polymorphic alleles of the already studied polymorphisms GSTM1 and GSTT1) 27. PCR is carried out as follows: first, the reaction mixture is heated to a temperature of 90-94 ° C, thereby causing DNA denaturation; then the temperature is reduced to 50-70 ° C, depending on the nucleotide sequence of the primers, so that annealing occurs in strictly complementary regions; finally, a temperature is set in the mixture that is optimal for the operation of DNA polymerase. When these cycles are repeated, the number of copies of the DNA site located between the primer sites increases exponentially. DNA amplification is carried out on a thermocycler using reagent kits for the detection of polymorphic markers of the GSTM1 gene and the GSTT1 gene.

5) Accounting for results. Amplification products are detected by gel electrophoresis and visualized using a transilluminating gel-documenting system. For the GSTM1 and GSTT1 genotypes, homozygous deletions were determined by the presence of one amplification fragment with a length of 350 bp, while heterozygotes and normal homozygotes were determined by two amplification fragments of a length of 480 and 350 bp.

The authors of the proposed method were examined patients with COPD I-II stages and COPD II and III stages associated with stable angina pectoris I-II FC, with phenotypic manifestations of the European race, of both sexes, living in the territory of the Primorsky Territory for more than 20 years. The diagnosis of COPD was established in accordance with the international GOLD classification of 2008 [1], the diagnosis of stable angina pectoris in accordance with national recommendations (2008).

According to the post-bronchodilation values of FEV _{1, the} patients were divided into groups: group I - 30 patients with stage I COPD (FEV ₁ more than 80% of the due), group II - 30 patients with stage II COPD (FEV ₁ - from 50 to 80% of the due ); Group III - 30 patients with stage II COPD (FEV ₁ - from 50 to 80% of the due), associated with IHD;

Group IV - 30 patients with stage III COPD (FEV ₁ = 30-49% of the due) associated with IHD. Exclusion criteria: myocardial infarction, stroke and other serious diseases; alcohol abuse persons of senile age and persons unable to understand the goals and objectives of the study; patient refusal to participate in the study.

The control group (n = 30) was formed from conditionally healthy non-smoking volunteers with normal values of FEV ₁ , FEV ₁ / FVC, AD, who are not relatives of the volunteers of the main group, and corresponding to the main group by age, gender, ethnicity, not having a history of pathology bronchopulmonary system, allergic and other chronic diseases.

The significance of differences in the distribution of allele and genotype frequencies between groups of patients and healthy individuals is assessed using the χ ² test using the BIOSTAT program. The differences were considered statistically significant at p less than 0.05. The relative risk of a disease for a particular symptom is calculated as the odds ratio (OR): OR = (a × d) / (b × c), where a is the frequency of the allele (genotype) in the patient sample, b is the frequency of the allele (genotype) in the control sample , c is the sum of the frequencies of the remaining alleles (genotypes) in the patient sample, and d is the sum of the frequencies of the remaining alleles (genotypes) in the control.

Among patients with stage I COPD, all were malicious smokers with PEI for more than 20 packs of years. Among patients with stage II COPD, 50% of the subjects were regular smokers with an emergency of more than 10 packs of years, 50% were heavy smokers with an emergency of more than 20 packs of years. Among smokers with no signs of COPD, 25% are non-regular smokers (MCP less than 10 pack-years), 36% - regular (MCP more than 10 pack-years), 39% - malicious smokers (MCP more than 20 pack-years). In the examined patients with COPD associated with coronary heart disease, the average smoking history, as well as the smoking index and the number of pack-years, significantly increased simultaneously with an increase in the severity of the disease and a decrease in FEV1 (p less than 0.05), respectively. This is also confirmed by the literature on a direct relationship between smoking intensity and lung function decline in smokers [25, 26]. The smoking person index (ICH) in patients was on average 153 (more than 140), the number of pack-years was 26.2 pack of years (more than 20), respectively, which indicates a high degree of nicotine-associated risk and indicates a high intensity of smoking.

Investigation of the GSTT1 gene polymorphism in COPD stages I-II. When comparing the frequency of homozygous deletion of the GSTT 1 gene, the following prevalence was revealed: in individuals without signs of COPD, 14%, in COPD stage I 14.2%, in stage II COPD 20%. At the same time, the relative risk of developing a more severe stage of COPD (stage II) increased by 1.5 times according to the odds ratio. However, the reliability of the above differences was not obtained (χ ² less than 2). When comparing the groups of “malicious smokers” in terms of the frequency of deletion of the GSTT1 gene, despite an increase in the relative risk of stage II COPD compared to non-ill smokers by 1.7 times, no significant differences were found either.

The study of the GSTT1 genotype in COPD associated with coronary artery disease showed that in the control group homozygous deletion was detected in 1 person, which amounted to 3.33%. In group III, the presence of a homozygous deletion of GSTT1 was observed in 2 patients (6.67%). The highest frequency of homozygous deletion of GSTT1 was recorded in group IV (almost every third patient). The difference in the frequency of occurrence of the deletion genotype GSTT1 between groups III and IV was significant (χ ² = 10, p = 0.01). Figure 1 and table 1 shows the distribution of the genotype by the presence / absence of a homozygous deletion of the GSTT1 gene in patients with COPD compared with the control.

Table 1 Genotype distribution by the presence / absence of a homozygous deletion of the GSTT1 gene in patients with COPD compared with the control Genotype GSTT1 / group Control (n = 30) Group III - COPD II, assoc. with coronary heart disease (n = 30) Group IV - COPD III, assoc. with coronary heart disease (n = 30) homozygous deletion 3.33% (n = 1) 6.67% (n = 2) 33.33% (n = 10) normal / heterozygous deletion 96.67% (n = 29) 93.3% (n = 28) 66.67% (n = 20)

The applied statistical method of OS showed that the zero genotype (homozygous deletion) of GSTT1 significantly increases the relative risk of COPD compared with group III and control (OS = 1.93, OS = 0.9, OS = 0.55, respectively).

The study of homozygous deletion of GSTM1 in COPD stage I-II. When comparing the frequency of homozygous deletion of the GSTM1 gene among the same groups, no significant differences were found.

A study of homozygous deletion of GSTM1 in COPD associated with coronary artery disease showed that homozygous deletion is observed in almost half (46.67%) of healthy individuals. The zero genotype (homozygous deletion) of GSTM1 is detected in a significantly larger number of cases in groups III and IV than in the control (56.67%, 60%, 46.67%, respectively; p = 0.01). The maximum number of homozygous deletions of GSTM1 was established in group IV and significantly exceeds the frequency of a similar genotype in healthy individuals by 13% and 3% in patients with stage III COPD (χ ² = 6, p = 0.01) (Fig. 2).

Figure 2 and table 2 presents the distribution of the genotype according to the presence / absence of homozygous deletion of the GSTM1 gene in patients with COPD compared with the control.

table 2 Genotype distribution by the presence / absence of a homozygous deletion of the GSTM1 gene in patients with COPD compared with the control Genotype GSTM1 / group The control
(n = 30) III group
COPD II, assoc. with coronary heart disease (n = 30) IV group
COPD III, assoc. with coronary heart disease (n = 30) homozygous deletion 46.67% (n = 14) 56.67% (n = 17) 60% (n = 18) normal / heterozygous deletion 53.33% (n = 16) 43.33% (n = 13) 40% (n = 12)

The relative risk of the disease (calculated as OS) was almost the same in groups III and IV and was approximately equal to unity (OS = 0.99, OS = 0.96%, respectively; p = 0.01). This indicates that there is an equal risk of developing COPD and IHD in the presence of homozygous division of GSTM1. Moreover, the presence of the zero genotype (homozygous deletion) of GSTM1, in contrast to the zero genotype (homozygous deletion) of GSTT1, is not associated with the severity of COPD.

When comparing the frequency of occurrence of deletions of the GSTT1 and GSTM1 genes in patients with stage I-II COPD and smokers without signs of COPD in general and, in particular, in smokers with PEI more than 25 pack-years, no significant differences were found. The presence of a combined zero genotype GSTM1 and GSTT1 (homozygous deletions for both genes) was observed in four patients (all patients from group IV), which leads to the complete absence of the corresponding enzymes for detoxification of tobacco combustion products.

Thus, the results obtained suggest a greater exposure of individuals carrying homozygous deletions of GSTM1 and GSTT1 to the risk of developing COPD in combination with coronary artery disease. Carriage of homozygous deletions found in the examined is more significant and can explain the adverse course of the disease associated with a violation of the second stage of neutralization of xenobiotics. The presence of a combined zero genotype GSTM1 and GSTT1 (homozygous deletions for both genes) was observed in four patients (all patients from group IV), which indicates a risk of developing a pathological condition in the form of a combined pathology - COPD associated with coronary heart disease - and significantly aggravates the condition.

The technical result provided by the claimed method consists in the possibility of predicting the individual risk of developing COPD associated with coronary heart disease before the development of the pathological process or in the early stages of the disease, as well as in reducing the complexity (calculate the ICP, NPL and set up a PCR reaction), invasiveness, and how consequence - safety and pain (only a single blood sampling is performed); there is no need for multiple studies (since the results do not change throughout life), there are no restrictions on the use of biological material, thus, the cost of examination is reduced in comparison with conventional clinical research methods.

The essence of the proposed method is illustrated by clinical examples of specific performance.

Example 1

Patient I., 44 years old, height 166 cm, weight 74 kg, with phenotypic manifestations of the European race, living in the territory of the Primorsky Territory for more than 20 years. Diagnosis: COPD stage II associated with stable angina of exertion FC II. ICH - 143 (more than 140), PEI - 21.6 pack-years (more than 20), which indicates a high degree of nicotine-associated risk (high smoking intensity). Examination of the genetic status revealed a homozygous deletion of GSTT1, which indicates the impossibility of biotransformation of the chemicals that make up cigarettes with the GSTT1 enzyme. The presence of a homozygous deletion of GSTT1 in combination with a high degree of nicotine-associated risk (according to ICH and EPL) indicates a high relative risk of a combined pathology - COPD associated with IHD, relative to the average population level. If appropriate measures are taken to reduce the impact of external adverse factors, the magnitude of this risk will be reduced.

Recommendations: 1. Complete cessation of smoking or gradual restriction of the number of cigarettes smoked per day. 2. Avoid inhalation of harmful, industrial gases, car exhausts, not be in smoky, dusty rooms, regularly ventilate your house; to be outdoors more.

Example 2

Patient N. 54 years old, height 168 cm, weight 75 kg, a patient with phenotypic manifestations of the European race, living in the territory of the Primorsky Territory for more than 20 years. Diagnosis: Stage III COPD associated with stable angina pectoris II FC. ICH - 153 (more than 140), PEI - 26.2 pack-years (more than 20), which indicates a high degree of nicotine-associated risk (high smoking intensity). When examining the genetic status, a combined homozygous deletion of GSTT1 and GSTM1 was found, which indicates the impossibility of biotransformation of the chemicals that make up cigarettes with GSTT1 and GSTM1 enzymes. The presence of homozygous deletions of GSTM1 and GSTT1 in combination with a high degree of nicotine-associated risk (according to ICH and EPL) indicates a very high relative risk of a combined pathology - COPD associated with IHD, relative to the average population level. If appropriate measures are taken to reduce the impact of external adverse factors, then the magnitude of this risk will be less.

Recommendations: 1. Complete cessation of smoking or gradual restriction of the number of cigarettes smoked per day. 2. Avoid inhalation of harmful, industrial gases, car exhausts, not be in smoky, dusty rooms, regularly ventilate your house; to be outdoors more.

Information sources

1. Global Initiative for Chronic Obstructive Lung Disease (GOLD). Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease. NHLBI / WHO workshop report. Last update 2008. www.goldcopd.org/.

2. Bova A.A., Lapitsky D.V. Modern approaches to the diagnosis and treatment of coronary heart disease in patients with chronic obstructive pulmonary disease. Medical news. - 2007. - No. 9. - S.7-14.

3. Huiart L., Ernst P., Suissa S. Cardiovascular morbidity and mortality in COPD. Chest 2005; 128: 2640-2646.

4. Kozlova L.I. Chronic obstructive pulmonary disease and coronary heart disease: some aspects of functional diagnostics. Pulmonology, 2001, v. 11, No. 2, 9-12.

5. Reference practitioner. Ed. A.I. Vorobyova, M .: Medicine, 1982, p.96-123.

6. Chronic obstructive pulmonary disease. M., CJSC Publishing House Binom, St. Petersburg., Nevsky dialect, 1998, p. 512.

7. Vasilkova T., Popova T. Medvedeva I. Metabolic syndrome and bronchial obstruction - two components of systemic inflammation // Doctor. 2008. No. 8, 19-21.

8. Vakhrushev Ya.M., Ermakov G.I., Sharaev P.N. Assessment of the metabolism of the main substance of connective tissue in chronic obstructive pulmonary disease // Ter. archive. 2006. 78 (3), 13-16.

9. Mironov G.E., Krivoshapkina Z.N., Velichkovsky B.T. Changes in the functional state of the liver during chronic obstructive bronchitis // Bulletin of the Russian Academy of Medical Sciences. 2004.3, 13-16.

10. Shaimeeva L.O. The role of metabolic disorders in patients with chronic bronchial obstruction // New St. Petersburg Medical Gazette. 2002.3, 91-93.

11. Shepelenko A.F., Mironov M.B. Sidorov Yu.O. Comprehensive treatment of exacerbations of chronic obstructive pulmonary disease // Attending Physician. 2006. No. 8, 14-16.

12. Shlyakhov U.I. Chronic obstructive pulmonary disease // Pulmonology, selected issues. 2001, No. 2, 1-9.

13. RU 2370773 C1, 10.20.2009.

14. RU 2359618 C1, 06.27.2009.

15. RU 2262889 C1, 10.27.2005.

16. RU 2382597 C1, 02.27.2010.

17. RU 2245550 C2, 01.27.2005.

18. RU 2210078 C2, 08/10/2003.

19. RU 2210076 C2, 08/10/2003.

20. RU 2356056 C2, 05.20.2009.

21. RU 2353931 C1, 04/27/2009.

22. RU 2161441 C2, 01/10/2001.

23. RU 2241226 C1, 11.27.2004.

24. RU 2224466 C1, 02.27.2004.

25. Imboden M. et al. Glutation S-transferase genotypes modify lung protection decline in the general population: SAPALDIA cohort study. Respir. Res. 2007; 8: 2.

26. Palma S. et al. Influence of glutation-S-transferase polymorphisms on genotoxic effects induced by tobacco smoke. Mutat. Res. 2007; 633 (1): 1-12.

27. Pemble S., Schroeder K.R., Spencer S.R. Meyer D.J., Hallier E., Bolt H.M., Ketterer B., Taylor J.B. Human glutathione S-transferase Theta (GSTT1): cDNA cloning and the characterization of a genetic polymorphism. (1994) Biochem. J. 300, 271-276.

28. Seidegard J., Worachek W.R., Pero R.W., Pearson W.R. Hereditary differences in the expression of the human glutathione transferase active on trans-stilbene oxide are due to a gene deletion (1988) Proc. Natl. Acad. Sci. USA, 85, 7293-7297.

29. Chan-Yeung M. et al. Polymorfisms of glutation-S-transferase genes and functional activity in smokers with and without COPD. Int. J. Tuberc. Lung dis. 2007; 11 (5): 508.

30. Jian-Qing He, Jian Ruan, John E. Conneff et al. Antioxidant gene polimorphisms and susceptibility to a rapid decline in lung function in smokers. Am. J. of Resp. and Crit. Care Med: 2002; 166: 323-328.

31. Ye Z., Song H., Higgins J.P.T. et al. Five glutation-S-transferase gene variants in 23,452 cases of lung cancer and 30,397 controls: meta-analysis of 130 studies. PLoS Med. 2006; 3 (4): 0524-0534.

32. Sram R.C. Effect of glutation-S-transferase Ml polymorfism on biomarkers of exposure. Environ. Hlth. Perspect. 1998; 106: 231-239.

33. Yanchina E.D., Ivtchik T.V. Associations of glutation-S-transferase M1 0/0 (GSTM1 0/0), GST1 0/0 and - 1562 C / T matrix metalloproteinase 9 (MMP9 CT) genotypes with a risk and clinical features of COPD. European Human Genetics Conference: Final Program and Abstracts. 2006.

34. Jeffery P.K. Remodelling and inflammation of bronchi in asthma and chronic obstructive pulmonary disease. Proceeding of the ATS 2004; 1: 176-183.

Claims (1)

A method for predicting an increase in the individual risk of chronic obstructive pulmonary disease associated with coronary heart disease, characterized in that nicotine-associated risk indicators, such as the smoking person index and the number of pack-years, are calculated, then blood is sampled, then DNA is extracted, then the method is isolated PCR analyzes the polymorphic loci of the GSTM1 and GSTT1 genes, then the amplification products are analyzed by electrophoresis, visualized using a transilluminating gel -documenting system and in individuals carrying homozygous deletions of GSTs with a high degree of nicotine-associated risk, they predict an increase in the relative risk of a combined pathology - chronic obstructive pulmonary disease associated with coronary heart disease.

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