CC BY
11DISTURBANCE OF CARBOHYDRATE AND LIPID METABOLISM IN PATIENTS WITH CORONARY HEART DISEASE AND OBESITY WITH DIFFERENT GENOTYPES OF GENE OF TUMOR NECROSIS FACTOR-a
(G - 308A)
Kravchun P.
Kadykova O.
Ryndina N.
Molotiagin D.
НАРУШЕНИЯ УГЛЕВОДНОГО И ЛИПИДНОГО ОБМЕНОВ У БОЛЬНЫХ С ИШЕМИЧЕСКОЙ БОЛЕЗНЬЮ СЕРДЦА И ОЖИРЕНИЕМ С РАЗНЫМИ ГЕНОТИПАМИ ГЕНА ФАКТОРА НЕКРОЗА ОПУХОЛИ-а (G-308A) Кравчун П.Г. Кадыкова О.И. Рындина Н.Г. Молотягин Д.Г.
ABSTRACT
The aim of the research. ESimation of di^urbance of carbohydrate and lipid metabolism in patients with coronary heart disease and obesity depending on different genotypes of the gene of tumor necrosis factor -a.
Materials and methods. The &udy enrolled 222 patients with coronary heart disease and obesity. The experimental group was made up by 115 patients with CHD whose body weight was within normal limits. The control group was represented by 35 apparently healthy people. Additionally, the patients with coronary heart disease and obesity were divided into subgroups depending on genotype of the gene of TNF-a (G-308A): group 1 was made up by carriers of А/А genotype (n=58), group 2 - by G/А genotype (n=90), group 3 - G/G genotype (n=74). The groups were compared according to age and sex. All patients underwent common clinical and in^rumental inve^igations. Statical processing of the findings was carried out by means of Stati^ica package, version 6.0 (StaSoftInc, USA).
Results. The analysis of indices of carbohydrate metabolism depending on genotypes of tumor necrosis factor-a gene in patients with coronary heart disease and obesity showed that carriers of A/A genotype had more dramatic disorders of carbohydrate metabolism in the form of hyperinsulinemia and reduced sensitivity of tissues to insulin, while patients with G/А and G/G genotypes are characterized by higher resi^ance to glucometabolic disorders. The obtained findings have made it possible to assume that A allel is a pathological form of polymorphism of tumor necrosis factor-a gene (G-308A) and G allel is characterized by protective action.
Conclusions. The analysis of indices of carbohydrate metabolism depending on genotypes of tumor necrosis factor -a gene in patients with coronary heart di-sease and obesity has shown that carriers of A/A genotype have more severe changes of carbohydrate metabolism in the form of hyperinsulinemia and decreased sensitivity of tissues to insulin, while patients with G/A and G/G genotypes are characterized by higher resi^ance to glucometabolic disorders. The principal feature of alteration of lipid metabolism in patients with coronary heart disease combined with obesity is Satirically significant hypertriglyceridemia associated with A/A genotype of polymorphism of G-308A of tumor necrosis factor-a gene.
АННОТАЦИЯ
Цель. Оценить нарушения углеводного и липидного обменов у больных с ишемической болезнью сердца и ожирением в зависимости от разных генотипов гена фактора некроза опухоли -a (G-308A).
Материалы и методы. Мы провели комплексное обследование 222 пациентов с ишемической болезнью сердца и ожирением. Группу сравнения составили 115 больных с ишемической болезнью сердца и нормальной массой тела. В контрольную группу вошли 35 практически здорових лиц. Дополнительно больных с ишемической болезнью сердца и ожирением были разделены на подгруппы в зависимости от генотипа гена фактора некроза опухоли-a (G-308A): в первую подгруппу вошли носители А/А генотипа (n=58), во вторую - G/А генотипа (n=90), в третью - G/G генотипа (n=74). Группы были сравнены по возрасту и полу. Всем пациентам проведено клиническое и инструментальное обследование. Статистический анализ полученных результатов проводили с использованием программы программного комплекса «StatiSica», версия 6,0 (StaSoftInc, США).
Результаты. Анализ показателей углеводного обмена в зависимости от генотипов гена фактора некроза опухоли-a у больных с ишемической болезнью сердца и ожирением показал, что носители генотипа А/А имеют более выраженные нарушения углеводного обмена в виде гиперинсулинемии и снижения чувствительности тканей к инсулину, тогда как пациенты с генотипами G/А и G/G обладают большей стойкостью к глюко-метаболическим нарушениям. Полученные данные позволяют допустить, что аллель А является патологическим вариантом полиморфизма гена фактора некроза опухоли-a (G-308A), а аллель G обладает протекторным действием.
Выводы. Анализ показателей углеводного обмена в зависимости от генотипов гена фактора некроза опухоли-a у больных с ишемической болезнью сердца и ожирением показал, что носители генотипа А/А имеют более выраженные нарушения углеводного обмена в виде гиперинсулинемии и снижения чувствительности тканей к инсулину, тогда как пациенты с генотипами G/А i G/G обладают большей стойкостью к глюко-метаболическим нарушениям. Ведущей особенностью перестройки липидного обмена у больных с ишемической болезнью сердца в сочетании с ожирением является статистически вероятная гипертриглицеридемия, которая ассоциирована с А/А генотипом полимор-физма G-308A гена фактора некроза опухоли-a.
Keywords: coronary heart disease, obesity, genotypes of gene of tumor nec-rosis factor-a.
Ключевые слова: ишемическая болезнь сердца, ожирение, генотипы ге-на фактора некроза опухоли-a.
Introduction. Obesity is an important risk factor of a range of serious medical challenges leading to reduced quality of life, significant increase of morbidity rate and untimely death in patients with coronary heart disease (CHD) [15].
Metabolic disorders associated with progression of CHD in patients with obesity were ftudied by many scientifls [1, 2, 8, 12, 13]. However, presymptomatic diagnosis due to assessment of gene polymorphism needs carrying out of further invefligations.
Purpose. Eflimation of diflurbance of carbohydrate and lipid metabolism in patients with coronary heart disease and obesity depending on different genotypes of the gene of tumor necrosis factor -a (G-308A).
Materials and methods. The fludy enrolled 222 patients with CHD and obesity who were undergoing treatment at Cardiology Department of CHPI (Communal Health Protection Inflitution) Kharkiv Municipal Clinical Hospital No 27 which is the base health facility of Department of Internal Medicine No 2, Clinical Immunology and Allergology of Kharkiv National Medical University of Miniflry of Health of Ukraine. Complete physical examination was provided for all of these patients. The experimental group was made up by 115 patients with CHD whose body weight was within normal limits. The control group was represented by 35 apparently healthy people. Additionally, the patients with CHD and obesity were divided into subgroups depending on genotype of the gene of TNF-a (G-308A): group 1 was made up by carriers of A/A genotype (n=58), group 2 - by G/A genotype (n=90), group 3 - G/G genotype (n=74). The groups were compared according to age and sex. The fludy did not involve patients with severe comorbi-dity of organs of respiration, digeflion, pathologies of the kidneys and people suffering from oncological diseases.
The diagnosis was made in accordance with valid orders of Miniflry of Health of Ukraine.
All patients underwent common clinical and inflrumental invefligations. In order to control carbohydrate metabolism, the level of glucose was assessed by means of glucose-oxidative method; eflimation of content of glycated hemoglobin (HbA1c) in the whole blood was carried out due to photometric technique through the reaction with thiobarbituric acid with use of the commercial tefl-syflem Reagent (Ukraine) according to the inflructions provided. Insulin concentration was assessed by means of immunoenzymometric method with the use of the commercial tefl-syflem INSULIN ELISA KIT produced by Monobind (USA). The insulin resiflance index of HOMA (Homeoflasis Model Assessment) was used. It was calculated according to the following formula:
insulin (mIU/ml) x glucose in fafling flate (mmol/L)/22.5. In case of HOMA index> 2.77 patients were considered to be insulin resiflant.
The biochemical fludy includes eflimation of level total choleflerol (TC) and high-density lipoproteins (HDL) by means
of the peroxidase method with the use of Choleflerol Liquicolor assay kit produced by Human (Germany) in the blood serum, flabilized by heparin. The level of triglycerides was assessed due to fermentation colorimetric method with the use of Triglycerides GPO assay kit produced by Human (Germany). The atherogenic index (AI) was calculated by means of A.M. Klimov's formula: AI= (TC - HDL)/HDL; the level of very low-density lipoproteins (VLDL) = triglycerides/2.2 x 0.45, (mmol/L); the level of low-density lipoprotein (LDL) = TC - (VLDL + HDL), (mmol/L).
The fludy was aimed to eflimate anthropometric indices of waifl measurement and hip width. In order to characterize obesity, the body mass index (Quetelet index) was assessed due to the following formula:
body mass (kg)/height (m2).
The fludy of polymorphous locus G-308A of TNF-a gene was carried out by means of the method of polymerase chain reaction with electrophoretic detection of findings with the use of SNP- EKCnPEC assay kit produced by LLC «Lytech» (Russian Federation). DNA purification out of the whole blood was performed
owing to the reagent «DNA-express-blood» produced by LLC «Lytech» (Russian Federation) according to the infractions provided. Accuracy of genotype frequency diflribution was assessed by means of correspondence of Hardy-Weinberg Equilibrium (pi2 + 2 pipj + pj2 = 1). In accordance with Declaration of Helsinki all patients were informed about clinical fludy and they gave their consent to assess polymorphism the gene in queflion.
Statiflical processing of the findings was carried out by means of Statiflica package, version 6.0. In order to compare diflribution of frequencies of alleles and genotypes between the groups, Pearson and Fisher's x2 criteria were used. To eflimate relative risk of development of diseases, odds ratio (OR) was calculated. As absence of associations OR=1 was considered; as a positive association - OR>1; as a negative association of allel or genotype with disease (low risk of disease development) OR<1 was considered. The confidence interval (CI) is an interval of va-lues which with a probability of 95% comprises prognoflic value of OR. Dif-ferences in p<0.05 were considered to be Satirically valid.
Findings and consideration. Due to the analysis of indices of carbohydrate metabolism it was revealed that in patients with CHD and obesity, carriers of A/A genotype of TNF- a gene (G-308A) the level of glucose was 4.57±0.11 mmol/L, insulin - 13.23±0.79 mcU/ml, glycated hemoglobin - 5.17±0.39 %; insulin resiflance index HOMA was 2.69±0.46 units; in carriers of G/A genotype - 4.50±0.09 mmol/L, 7.63±0.68 mcU/ ml, 5.09±0.28 %, 1.53±0.35 units respectively; in case of G/G genotype the abovementioned indices corresponded to the following values: 4.46±0.08 mmol/L, 7.11±0.80 mcU/ml, 4.97±0.34 % and 1.41±0.43 units. (Table 1).
Table 1
Indices of carbohydrate metabolism in patients with CHD and obesity depending on genotypes of TNF- a gene (G-308A) (M±m)
Groups Indices Genotypes of TNF-a gene (G-308A) P
А/А (n=58) G/А (n=90) G/G (n=74)
НОМА, un. 2.69±0.46 1.53±0.35 1.41±0.43 р1-2<0.001 р1-3<0.001 р2-3>0.001
HbA1c,% 5.17±0.39 5.09±0.28 4.97±0.34 р1-2>0.001 р1-3>0.001 р2-3>0.001
Blood glucose, mmol/L 4.57±0.11 4.50±0.09 4.46±0.08 р1-2>0.001 р1-3>0.001 р2-3>0.001
Insulin, mcU /ml 13.23±0.79 7.63±0.68 7.11±0.80 р1-2<0.001 р1-3<0.001 р2-3>0.001
Comparison of indices of carbohydrate metabolism depending on different genotypes of TNF-a gene (G-308A) in patients with CHD and obesity showed, that in patients with A/A genotype the values of insulin and insulin resi^ance index HOMA were significantly higher. Insulin level was higher in patients with A/A genotype up to 42.33 % and 46.26 % than in patients with G/А and G/G genotypes and insulin resi^ance index HOMA - 43.12 % and 47.58 % respectively (р<0.001). Concerning the levels of glucose and glycated hemoglobin, significant differences depending on genotypes of TNF-a gene were not revealed (р>0.001).
Therefore, the analysis of indices of carbohydrate metabolism depending on genotypes of TNF- a gene in patients with CHD and obesity showed that carriers of A/A genotype had more dramatic disorders of carbohydrate metabolism in the form of
hyperinsulinemia and reduced sensitivity of tissues to insulin, while patients with G/А and G/G genotypes are characterized by higher resi^ance to glucometabolic disorders. The obtained findings have made it possible to assume that A allel is a pathological form of polymorphism of TNF-a gene (G-308A) and G allel is characterized by protective action.
As it is seen from Table 2, wai& measurement, hip width as well as their ratio were not different in patients with CHD and obesity depending on genotypes of TNF- a gene (G-308A) (р>0.05). Body mass index in carriers of A/A genotype, in its turn, had the higher value (39.43±0.62 kg/m2) that was up to 17.93 % and 18.49 % more than in carriers of G/A and G/G genotypes (р<0,05). Consequently, A/A genotype in patients with CHD and obesity was associated with body mass index.
Table 2
Status of body type indices in patients with CHD and obesity depending on genotypes of TNF- a gene (G-308A) (M±m)
Groups Indices Genotypes of TNF-a gene (G-308A) P
А/А (n=58) G/А (n=90) G/G (n=74)
Waist measurement, cm 114.89±1.57 113.64±1.22 112.89±1.35 р1-2<0.05 р1-3<0.05 р2-3>0.05
Hip width, cm 114.12±1.63 113.16±1.23 112.03±1.56 р1-2>0.05 р1-3>0.05 р2-3> 0.05
Waist measurement/hip width 1.01±0.001 1.00±0.001 1.01±0.002 р1-2>0.05 р1-3>0.05 р2-3>0.05
Body mass index, kg/ m2 39.43±0.62 32.36±0.54 32.14±0.58 р1-2<0.05 р1-3<0.05 р2-3>0.05
Neck width, cm 59.34±0.98 48.67±0.84 49.13±1.11 р1-2<0.05 р1-3<0.05 р2-3>0.05
Making an assessment of lipid metabolism in patients with CHD and obesity, firfl of all, it should be noted that all indices exceeded flandard values. It is due to influence of CHD and obesity on alteration of lipid metabolism caused by pa-thogenic agents (Table 3).
Significant differences concerning the levels of total choleflerol, HDL choleflerol, LDL choleflerol, VLDL choleflerol
and AI depending on genotypes of TNF-a gene (G-308A) in patients with CHD and obesity were not revealed (р>0.05). The level of total chole^erol was within the range from 5.47±0.08 mmol/L to 5.63±0.06 mmol/L; HDL chole^erol was 0.85±0.03 mmol/L in carriers of A/A genotype, 1.13±0.04 mmol/L in carriers of G/А genotype and 1.26±0.03 mmol/L in people with G/G genotype, LDL chole^erol - 0.85±0.03 mmol/L, 1.13±0.04
mmol/L and 1.26±0.03 mmol/L, VLDL chole^erol - 1.89±0.05 mmol/L, 1.82±0.03 mmol/L and 1.80±0.04 mmol/L and AI -4.84±0.08, 4.76±0.06 and 4.68±0.07 respectively.
The level of triglycerides in the group of patients with A/A genotype was significantly higher up to 30.80 % and 33.33 %
Indices of lipid metabolism in patients with CHD and obesity
than in patients with G/A and G/G genotypes. It was 2.37±0.08 mmol/L vs 1.64±0.07 mmol/L and 1.58±0.09 mmol/L (p<0.05).
Thus, the principal feature of alteration of lipid metabolism in patients with CHD combined with obesity is Satirically significant hypertriglyceridemia associated with A/A genotype of polymorphism of G-308A of TNF- a gene.
Table 3
depending on genotypes of TNF- a gene (G-308A) (M±m)
Groups Indices Genotypes of TNF-a gene (G-308A) P
А/А (n=58) G/А (n=90) G/G (n=74)
Total cholesterol, mmol/L 5.63±0.06 5.60±0.07 5.47±0.08 р1-2<0.05 р1-3<0.05 р2-3>0.05
Triglycerides. mmol/L 2.37±0.08 1.64±0.07 1.58±0.09 р1-2>0.05 р1-3>0.05 р2-3> 0.05
HDL cholesterol, mmol/L 0.85±0.03 1.13±0.04 1.26±0.03 р1-2>0.05 р1-3>0.05 р2-3>0.05
LDL cholesterol, mmol/L 3.54±0.06 3.49±0.07 3.45±0.09 р1-2<0.05 р1-3<0.05 р2-3>0.05
VLDL cholesterol, mmol/L 1.89±0.05 1.82±0.03 1.80±0.04 р1-2<0.05 р1-3<0.05 р2-3>0.05
AI 4.84±0.08 4.76±0.06 4.68±0.07 р1-2<0.05 р1-3<0.05 р2-3>0.05
The fludy of character of interrelations between the indices which were discovered according to genotypes of TNF-a gene in patients with CHD and obesity are represented in Table 4. Direct correlation relationships between A/A genotype and the level of insulin (r=0.56, p<0.05), insulin resiflance index
Matrix of intercorrelations between the indices of carbohydi with CHD and obesity (rcrit=0.24)
HOMA (r=0.43, p<0.05), body mass index (r=0.71, p<0.05), triglycerides (r=0.69, p<0.05) were defined.
The obtained data are indicative of involvement of polymorphous locus G-308A of TNF-a gene into formation of disorders of carbohydrate and lipid metabolism that corresponds to the literature.
Table 4
s ad lipid metabolism and genotypes of TNF-a gene in patients
Genotype Index А/А G/А G/G
Glucose 0.22 0.22 -0.13
Insulin 0.56* 0.21 -0.18
НОМА 0.43* 0.20 -0.20
HbAlc 0.14 0.14 -0.21
Body mass index 0.71* 0.22 -0.34*
Waist measurement 0.21 0.18 -0.09
Hip width 0.16 0.09 -0.10
Waist measurement / hip width 0.20 0.07 -0.20
Total cholesterol 0.13 0.16 -0.23
Triglycerides 0.69* 0.15 -0.16
HDL cholesterol 0.21 0.22 0.14
LDL cholesterol 0.09 0.16 -0.17
VLDL cholesterol 0.18 0.21 -0.16
AI 0.22 0.22 -0.18
Note: *р<0.05, rcrit=0.24
Thus, Sookoian S.C. et al. [10] flates that TNF-a gene participates in pathogenesis of metabolic syndrome and can increase risk of its development. In this case in individuals who have A allel of TNF-a gene, the risk of development of obesity is 23%higher, the level of insulin in the blood plasma in fafling flate is by 1.25 times higher in comparison with G/G homozygotes. According to logiflic regression analysis, in carriers A allel of TNF-a gene the risk of development of hypertensive disease depending on body mass index, waifl measurement/hip width ratio is by 2.8 times higher. According to the data provided by Fontaine-Bisson B. et al. [3] among the patients with obesity, carriers of A allel of TNF-a gene had higher resiflance to insulin. At the same time, according to the fludy carried out by Popko R. et al. [4], in children with overweight genetic -308A of TNF-a gene was recorded more frequently in comparison with the control (p=0.04). Furthermore, many scientifls note that concentration of TNF-a tends to increase in case of obesity [14]. According to meta-analysis carried out by Yu S. et al. [6], polymorphism-308G>A of TNF-agene is a risk factor of development of diabetes mellitus of type II in population of China. The same associations were eflablished in Asian population [11]. However, in other ethnic group, e.g. in Caucasians [5], Japanese [9] and Indians [7] significant relations of -308G/A of TNF-a with formation of diabetes mellitus were not eflablished.
Conclusions
1. The analysis of indices of carbohydrate metabolism depending on genotypes of TNF-a gene in patients with CHD and obesity has shown that carriers of A/A genotype have more severe changes of carbohydrate metabolism in the form of hyperinsulinemia and decreased sensitivity of tissues to insulin, while patients with G/A and G/G genotypes are characterized by higher resiflance to glucometabolic disorders.
2. The principal feature of alteration of lipid metabolism in patients with CHD combined with obesity is Satirically significant hypertriglyceridemia associated with A/A genotype of polymorphism of G-308A of TNF- a gene.
Reference.
1. Dawber, T. R. Incidence of coronary heart disease, flroke and peripheral arterial disease in the Framingham Study: the Epidemiology of Atherosclerotic Disease / T. R. Dawber. -Cambridge : M. A. Harvard University Press, 1980. - P. 59-75.
2. Fernandez-Real, J. M. Insulin resiflance and chronic cardiovascular inflammatory syndrome / J. M. Fernandez-Real, W. Ricart // Endocr. Rew. - 2003. - Vol. 24. - P. 278-301.
3. Genetic polymorphisms of tumor necrosis factor-alpha modify the association between dietary polyunsaturated fatty acids and plasma high-density lipoprotein-choleflerol concentrations in a population of young adults./ B. Fontaine-Bisson, A. El-Sohemy //J Nutrigenet Nutrigenomics. - 2008. -Vol. 1(5). - P.215-223.
4. Influence of proinflammatorycytokine gene polymorphism on childhood obesity / K. Popko, E. Gorska, B. Pyrzak , et al. // Eur. J. Med. Res. - 2009. -Vol. 14, suppl. 4. - P. 59-62.
5. Large-scale association analysis of TNF/LTA gene region polymorphisms in type 2 diabetes. / V. Boraska, N.W. Rayner, C.J. Groves, et al.// BMC Med Genet. - 2010. - Vol.11. - P.69.
6. Metabolic syndrome in hypertensive adults from rural NortheaS China: an update. / S. Yu, X. Guo, H. Yang, L. Zheng, Y. Sun // BMC Public Health. - 2015. - Vol. 15. - P.247.
7. Mеtаgеnоmiс &udy оf singlе-nuсlеоtidе роlymоrрhism within саndidаtе gеnеs аssосiаtеd with tyре 2 diаbеtеs in аn Indiаn рорulаtiоn[Tехt] / Р. N.Mukhораdhyаyа, А.Асhаryа, Y.Сhаvаn[еt а1.] // Gеnеt. Mоl. Rеs. - 2010. - Ш. 9, № 4. - Р. 2060-2068.
8. Neaton, J. D. Serum chole^erol, blood pressure, cigarette smoking, and death from coronary heart disease. Overall findings and differences by age for 316099 white men (Multiple Risk Factor Intervention Trial Research Group) / J. D. Neaton, D. Wentworth // Arch. Intern. Med. - 1992. - Vol. 152. - P. 56-64.
9. Relationship of the tumor necrosis factor-alpha -308 A/G promoter polymorphism with insulin sensitivity and abdominal fat di^ribution in Japanese patients with type 2 diabetes mellitus. / M. Furuta, Y. Yano, K. Ito, et al. // Diabetes Res Clin Pract. -2002. - Vol. 56(2). - P.141-145.
10. Sооkоiаn, S. С. Mеtа-аnаlysis оп Ше G-308A tumоr xerosis fасtоr а^а gеnе vаriаnt аМ рhеnоtyреs аssосiаtеd with Ше mеtаbоliс syndrоmе [Тех^] / S. С. Sооkоiаn, С. Gоnzalеz, С. J. РпЫа // Оbеs. Rеs. - 2005. - Ш. 13, № 12. - Р. 2122-2131.
11. Tumоr xerosis fасtоr-аlрhа G-308A gеnе роlymоrрhism аМ соrоnаry hеаrt disеаsе susсерtibility: аи uрdаtеd mеtа-аnаlysis ^xt] / H. F. Zhаng, S. L. Х1е, J. F. Wаng fct а!] // Thromb. Rеs. - 2011. - Ш. 127, № 5. - Р. 400-405.
12. Аметов А.С., Демидова Т.Ю., Целиковская А.Л. Ожирение и сердечно-сосудистые заболевания // Тер. архив. -2001. - № 8. - С. 69-72.
13. Маньковский, Б. Н. Роль гипергликемии в развитии микрососудистых и кардиоваскулярных осложнений сахарного диабета / Б. Н. Маньковский // Лжи Украни. - 2010. - № 6. - С. 11-15.
14. Урбанова, К. А. Изменение чувствительности к инсулину у лиц с различными нарушениями углеводного обмена [Текст] / К. А. Урбанова, Г. Р. Галстян // IV Всероссийский диабетологический конгресс, Москва, 19-22 мая 2008 г. : тез. докл. / М-во здравоохранения и соц. развития Рос. Федерации, Федер. агентство по высокотехнол. мед. помощи, Эн-докринол. науч. центр Росмедтехнологий [и др.]. - Москва, 2008. - С. 81.
15. Фадеенко Г. Д. Роль ожиршня та цукрового дiабету 2 типу у виникненш атеротромботичного ризику при iшемiч-нш xворобi серця / Г. Д. Фадеенко, О. £. Запровальна. // Украшський терапевтичний журнал. - 2014. - № 3-4. - С. 28-36.
