CC BY
15
In both groups, there was observed a significant increase of TNF-a in blood serum comparatively to the control group (p <0.05). The largest increase in 2.5 times (p <0.001) was observed in concomitant course of AH and DM type 2.
CRP levels in blood serum exceed the reference values in both groups of surveyed patients (p <0.05). The greatest increase for CRP indicators (in 2.2 times) was observed in patients with comorbidity (p <0.05) and correlated with BMI (r = 0,43; p <0.00l), FBG level (r = 0,48; p < 0.001) and TG levels (r = 0,37; p <0.04), index of HOMA-IR (r = 0,46; p <0.00l).
Conclusion. It was found that the mechanisms of metabolic disorders in patients with concomitant AH and DM type 2, characterized by insulin resistance and progression of the development of atherogenic dyslip-idemia, an increase in markers of systemic inflammation are most pronounced in patients with overweight and obesity.
Thus, a comprehensive diagnosis ofAH and DM type 2, will contribute to the individualization of preventive and therapeutic measures, as well as the establishment of control for the progression of atherosclerosis and reduce cardiovascular risk.
References:
1. Inflammation in the pathophysiology of essential hypertension/F. Montecucco A., Pende A. Quercioli [et al.]//J. Nephrol. - 2011. - Vol. 24. - P. 23-34.
2. Shimamoto K. Metabolic syndrome/K. Shimamoto, T. Miura//Nippon Rinso. - 2015. -V. 67 (4). - P.771-776.
3. Yamawaki H. Vascular effects of novel adipocytokines: focus on vascular contractility and inflammatory responses/H. Yamawaki//Biol. Pharm. Bull. - 2011. - Vol. 34 (3). - P. 307-310.
4. Hackam D. G. The 2010 Canadian Hypertension Education Program recommendations for the management of hypertension/Hackam D. G. et al.//Can. J. Cardiol. - 2014. - Vol. 26, - No 5. - P. 249-258.
5. Sarwar N., Gao P., Seshasai S. R., et al. Diabetes mellitus, fasting blood glucose concentration, and risk ofvascular disease: a collaborative meta-analysis of 102 prospective studies/N. Sarwar, P. Gao, S. R. Seshasai, et al.//Lan-cet. - 2016. - Vol. 375 (9733). - P. 2215-22.
6. Pereira M., Lunet N., Azevedo A., Barros H. Differences in prevalence, awareness, treatment and control of hypertension between developing and developed countries/M. Pereira et al.//J. Hypertens. - 2012. - Vol. 27. - P. 963-975.
7. Fonseca V. A. Defining and Characterizing the Progression of Type 2 Diabetes/V. A. Fonseca//Diabetes Care. -2009. - Vol. 32 (2). - P. 151-56.
8. Arror A. R. Insulin resistance and heart failure: molecular mechanisms/A. R. Arror//Heart Fail Clin. - 2012. -Vol. 8 (4). - P. 3133-3140.
9. Boudina S. Diabetic cardiomyopathy, causes and effects/S. Boudina//Rev. Endocr. Metab. Disord. - 2015. - No 11. - P. 31-39.
10. Zhou J. and Qin G. Adipocyte dysfunction and hypertension/J. Zhou and G. Qin//American Journal of Cardiovascular Diseas. - 2012. - Vol. 2. - P. 143-149.
DOI: http://dx.doi.org/10.20534/ELBLS-17-1-16-19
Lytvynets Liudmila Yaroslavivna, Doctor of Medicine, Professor, Departament of Pediatrics Ivano-Frankivsk National Medical University E-mail: [email protected]
Clinical significance of gene polymorphism of xenobiotic Detoxification in children with bronchial asthma
Abstract: The phenotypic peculiarities of bronchial asthma (BA) in 94 children from Ivano-Frankivsk region with different types of allelic gene polymorphism of the first phase of xenobiotic detoxification — mEPXHl have been investigated. The association between polymorphism of gene mEPXHl and susceptibility
to occurrence and severity of BA in children have been described. It has been established that the manifestation of asthma as eco-determined pathology is affected by individual types of metabolic activity of genes of xenobiotic detoxification. It has been proved that patients with uncontrolled asthma have genotypes CC and GG (p <0.05) much more often. Children with asthma often have high enzymatic activity of a gene mEPHXl (polymorphic variants of AG and AA-TT-TT). A "slow" functional state of gene mEPHXl corresponding to homo- and hetero-zygotes for the mutant alleles: CC-AA and TC-AA is also a frequent phenomenon. It has been established that bronchial asthma as eco-determined pathology is affected by individual variants of metabolic activity of genes of xenobiotic detoxification.
Keywords: bronchial asthma, children, genes of xenobiotic detoxification, polymorphism of genes.
Introduction. Identification of interacting specific The aim of our research was to study the phenotypic
genes and exogenous factors which form human resis- characteristics of asthma in Pre-Carpathian children with
tance to the environment is of considerable interest for clinical medicine. However, the range of reactions of different individuals to the same type of the environmental influences is rather variable, ranging from preservation of health in some people to the development of serious diseases in others. If the same effects lead to different reactions, this poses the question of causal factors of such selectivity. It is known that children are the most susceptible to harmful external factors due to the incomplete development of all functional body systems. Therefore, due to unfavourable ecological situation the problem of influence of genetic and environmental factors on the pathogenesis of chronic respiratory diseases in children and its investigation is topical today. BA is a leader among bronchial and pulmonary diseases caused by the influence of pneumo-tropic pollutants. The enzymes of xenobiotic biotransformation system play an important role in protecting the lungs from toxic substances [1; 3; 5; 11]. Biotransformation consists in the modification of xenobiotic physical properties from lipophilic to hydrophilic that facilitates their removal from the body. Protection system from xenobiotics is regulated by genes of xenobiotic detoxification (GXD) and presented by three-stage process, including the first phase of activation, the second phase — the neutralization, and the third phase — excretion ofxenobiotics from the body [2; 4; 5; 6]. Entering the body xenobiotics are activated by enzymes of cytochromes P-450, flavin-linked mono-oxygenases, esterases, aldehyde dehydrogenases and microsomal epoxide hydrolases which participate in the reactions of oxidation, restoring and hydrolysis of xenobiotic molecules, forming the intermediate electrophilic metabolites — soluble compounds possessing genotoxic properties. All these enzymes are localized in the membranes of the endoplasmic retic-ulum and belong to microsomal or monooxygenase system or metabolism [6; 7; 9; 12].
different types of allelic gene polymorphism of the first phase of xenobiotic detoxification of microsomal epoxide hydrolases (mEPHXl).
Materials and methods. The study involved 94 children aged 6 to 18 years, patients with BA who were hospitalized in Ivano-Frankivsk Regional Pediatric Hospital at Allergic department during 2009-2010. The diagnosis of asthma was verified in accordance with the Protocol of diagnosis and treatment of BA in children № 868 from 03.10.2013. According to the level of controllability for asthma, taking into account the results of BA control test (GINA, 2009) the children were divided into two groups: 44 patients (46.8%) — with partially controlled asthma (PCBA), 50 patients (53.2%) — with uncontrolled asthma (UCBA). The control group consisted of 157 children who live in different parts of Ivano-Frankivsk region, selected by random sampling. The material of genetic investigations was DNA isolated from leukocytes of patients' peripheral blood. Isolation and purification of DNA was carried out using commercial kits DIAtom TM DNA Prep200, GenePak DNA PCR test (LLC "Laboratory IzoGen") according to the manufacturer's recommendations. Polymorphic variants of a gene mEPHXl were identified using multilocal PCR . Mathematical processing of the results was performed using the statistical program Statistica.
Results and discussion. During the investigation we analyzed gene polymorphism of the first phase of xenobiotic detoxification in general sampling of patients with asthma and in control group. The complex analysis of gene polymorphism encoding enzymes of the first phase of xenobiotic biotransformation, i. e. mEPHXl discovered a number of consistent patterns (Table 1).
A comprehensive analysis of gene mEPHXl polymorphism in healthy children and in 94 patients with asthma revealed a number of regularities. Thus, the most common deletions among healthy children are TC (64) (41.0%) and TT (69) (44.0%) and AA (99) (63.0%)
Table 1. - The distribution of gene mEPHXIpolymorphism in healthy children and children with asthma
Genotype variants UCBA 1 (n=50) PCBA2 (n=44) P1-2 Healthy3 (n=157) P1-3 P2-3
Genotype mEPHXl T337 C
TC 14 (31,8%) 23 (46,0%) 0,08 64 (41,0%) 0,042 0,26
TT 21 (47,7%) 21 (42,0%) 0,19 69 (44,0%) 0,33 0,40
CC 9 (20,5%) 6 (12,0%) 0,06 24 (15,0%) 0,21 0,28
Genotype mEPHXl A415G
GG 6(13,6%) 5 (10,0%) 0,12 13 (8,0%) 0,001 0,354
AG 17 (38,8%) 21 (42,0%) 0,29 46 (29,0%) 0,051 0,047
AA 21 (47,6%) 24 (48,0%) 0,31 99 (63,0%) 0,001 0,029
Remarks:
1. All data are presented in absolute numbers
2. Percentage of people with a particular genotype of the total number of children in group is given in brackets
3. P - probability determined by quantity comparison method
Analysis of frequency combinations of genotypes mEPHXl ofxenobiotic detoxification gene mEPHXl in children with varying degrees of BA controllability proved that phenotypic variant of asthma is largely determined by the state of genotype.
Thus, homozygous mutant deletion of CC (His113His) in patients with PCBA was in 6 cases, and in patients with UCBA in 9 cases, i. e. 8.5% more common in the children with UCBA.
Comparison of frequencies of alleles and genotypes TC (Tyr113His) in patients with varying degrees of asthma control revealed that in children with UCBA the proportion of hetero-agents was 14 (31.8%) versus 23 (46.0%) in children with PCBA; a significant difference with the same rate was observed in healthy children (Pn<0.05).
The frequency of the normal homozygous allele TT (Tyr113Tyr) in children with different phenotypes of asthma and healthy too, did not differ significantly. As for availability of polymorphic variants of the locus A415G gene mEPHX1 in children with varying degrees of the disease control, the following data were obtained: in children with UCBA a homozygous GG variant was noticed in 6 cases, and its frequency was significantly higher than in healthy children (PN<0.05).
It is known that the prevalence of homozygous GG variant indicates the increased activity of the enzymes encoded by the gene mEPHX1 to 20.0%.
Thus, in children with a severe asthma, despite high enzymes activity, the severity of illness is defined by a low activity of gene products of the second phase of xenobi-otic detoxification. In such patients the increased activity of enzymes encoded by the gene mEPHX1, induces the accumulation of active intermediates, contributes to
oxidative stress and, as a result the disease.
Homozygous variant AA (His139 His) in children with PCBA and UCBA was almost at the same level: 24 (48.0%) and 21 (47.6%) respectively, and occurred much less frequently than in healthy children 99 (63.0%) (Pn<0.05).
Heterozygous variant AG (His139Arg) more often, i. e. in 21 (42.0%) was observed in children with PCBA, but the difference was not evidental. At the same time, this variant occurred in 46 healthy children, that is more often than in patients with asthma (Pn<0.05).
Thus, genetic testing of children with BA on the basis of genes detoxification of xenobiotics of the first phase and their combinations can detect in the genome the tendencies to BA appearance and development of the disease of certain degree of controllability.
Prospects for further research. Implementation of diagnostic methods of studying genetic markers in children with asthma will allow predicting the severity of the disease and enabling allergists, pediatricians, and family doctors to identify ways to early prevention of the disease.
Conclusions
1. The realization of asthma as eco-determined pathology is influenced by individual variants of metabolic activity of genes of xenobiotic detoxification.
2. Participation of genes of xenobiotic metabolism in modulating process of mutagenesis intensity of pathogenetically severe forms of BA suggests their possible contribution to the progression of the disease.
3. In children with inherited weak genotype the inac-tivation of xenobiotics is particularly slow and therefore the circumstances for harmful influence of metabolites on the body are especially realistic.
References:
1. Akhmadishina L. Z. Gene polymorphism of cytochromes - P 450, (CYP1A1, CYP1A2, CYP2E1) and the risk of occupational chronic bronchitis/L. Z. Akhmadishina, G. F. Korytina, O. V. Kochetova et al.//Med. Genetics. - 2007. No 6 (7). - P. 32-37.
2. Bragina E.Yu. Gene polymorphism of biotransformation of xenobiotics GSTT1, GSTM1, CYP2E1 and CY-P2C19 in patients with atopic asthma/E.Yu. Bragin, M. B. Freidin, I. A. Ten, L. M. Ogorodova//Bulletin of RAMS. - 2005. No 3 (117). - P. 121-125.
3. Baranov V. S. Testing of detoxification system genes in the prevention of certain multifactorial diseases/V. S. Ba-ranov, T. E. Ivaschenko, Baranova E. V.//Journal of Obstetrics and women's diseases. - 2003. - No 2 Vol. LII. - P. 11-16.
4. Vishtak N. V. Allelic polymorphism of the gene mEPHX as a marker of tendency to forming environmentally determined state of children/N. V. Vishtak, O. Z. Hnateiko//Environment and Health. - 2011. - No 2, 957. - P. 15-19.
5. Kononykhina N. V. Engagement of polymorphic variants of the gene EPHX1 in the development of chronic pulmonary disease of professional and non-professional origin among the population of Kursk region/N. V. Kononykhina, O. N. Bachinskiy V. I. Babkina et al.//Pulmonology. - 2011. - No 5. - P. 25-28.
6. Mazurina S. A. Role of polymorphism of genes-candidates in the development of atopic asthma/S. A. Mazurina, V. A. Kaznacheev//Russian allergological journal. - 2011. - No 3. - P. 14-22.
7. Ogorodova L. M. Significance of genetic predicts for the primary prevention of asthma in children with atopic dermatitis/L. M. Ogorodova, O. S. Fedorova, E.Yu. Bragina et al.//Pediatrics named after Speranskiy. - 2005. -No 6. - P. 4-6.
8. Puzyrev V. P., Stepanov V. A., Freidin M. B. The molecular basis of common multifactorial diseases/V. P. Puzyrev, V. A. Stepanov, M. B. Freidin. Genomics - medicine//Ed. by academician of RAMS V. I. Ivanov and academician L. L. Kiselyov. - M.: ICC «Akademkniga», - 2005. - 392 p.
9. Tyumentseva E. S. Using molecular and genetic methods for studying genetic bases of susceptibility to atopic diseases in children/E. S. Tyumentseva, N. V. Petrova, I. I. Balabolkin//Russian allergological journal. - 2011. -No 3. - P. 48-55.
10. Freidin M. B. Syntropic genes of allergic diseases/M. B. Freidin, V. P. Puzyrev//Genetics. - 2010. - Vol. 46. - No 2. - P. 224-229.
11. Fryer A. A. Polymorphism at the glutathione S-transferase GSTP1 locus. A new marker for bronchial hyper-responsiveness and as London S. J. Gene-air pollution interactions in asthma/S. J. London//Proc. Am. Thorac. Soc. - 2007. - V 3. - P.217-220.
12. Garcia-Aymerich j. Phenotypic heterogeneity of chronic obstructive pulmonary disease./J. Garcia-Aymerich, A. Agusti, J. A. Barbera et al.//Arch. Bronconeumol. - 2009. - V. 45. - S. 93. - P. 133-142.
13. Just J. Air pollution and asthma in children/J. Just, L. Nisakinovic, Y. Laoudi, A. Grimfeld//Arch Pediatr. -2006. - V. 7. - P. l055-1060.
14. London S. J. Gene-air pollution interactions in asthma/S. J. London//Proc. Am. Thorac. Soc. - 2007. - V 3. -P. 217-220.
