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14THE INTEGRATIVE SYSTEM OF BIOELEMENTS, IMMUNOLOGICAL RESISTANCE, ENZYME AND HORMONAL ACTIVITY OF ATHLETES IN CONDITIONS OF DEVELOPMENT OF LOCAL REGIONAL MUSCLE ENDURANCE IN LOWLAND AND MIDLAND IN DIFFERENT SEASONS OF THE YEAR
A.P. Isaev, professor, Dr.Biol., honorary figure of science of the Russian Federation A.S. Aminov, associate professor, Ph.D. V.V. Erlikh, associate professor, Ph.D. A.V. Nenasheva, associate professor, Dr.Biol.
South Ural state university (SCU), Institute of sport, tourism and service, Chelyabinsk
Key words: phases of adaptation, stress, system-structural mark, human elemental status, humoral and cell-bound immunity, enzyme and hormonal activity, midland, local-regional muscle endurance, system of intellectual analysis of physiological researches of athletes.
Introduction. The integration of the sport training theory and the adaptation theory is of great importance, since the accumulated huge amount of information demands new approaches for analysis and interpretation. The problem of estimation in sport includes various aspects of monitoring, specific certification, variability of some components of the basic and supporting systems of simulation and prediction, correction of conditions, fitness, recovery and regulating integral functions of the athletes' organisms. The scientifically based modification of efficient techniques of athletes' training is required.
Modern science provides a wide range of information in which the conceptions of many researchers are lost. Therefore, in the "Energy and resource conservation (NDP-5)" government project we focused on the development of a system for analysis of physiological examinations of elite athletes [2].
The purpose of the study was to substantiate the integrative system of order, self-regulation, interaction of metabolic processes in conditions of hypoxia at sports activities.
Materials and methods. The resistance to hypoxia was trained both by breath holding in relative rest conditions, and by short-term motor exercises aimed at particular endurance during mountain acclimatization. We combined the efforts of scientists, trainers and athletes, and obtained promising sport results. The efforts were focused on the development of local and regional muscle endurance (LREM, 50% training time) during the preparatory stages of training, combined with relaxation, stretching, swimming, sauna and massage; complex diagnostic control was applied: noninvasive screening hemogram analyser AMP, 3D spinal scanner, 3D stabilograph, urine analyzer, Oxycon Mobile telemetric system (Germany), Schiller ergospirometry (Switzerland, Germany), Tanita body composition analyser (Japan) and estimation of immunological parameters.
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We inspected 19-22 year old middle-distance cross-country skiers and wrestlers (judokas and sambo wrestlers) (ranked international masters of sports, masters of sports and candidates master of sports). The results were processed using up-to-date techniques.
The concentration of bioelements in biosubstrates corresponded to the content of chemical elements in the body and to their metabolism [9, 10]. The neuroendocrine integrations influence oxidation processes, metabolism, immunologic resistance and the determination of the interrelation between experimental data and enzyme and hormonal activity. One can assume that the integrative activity of the body is determined by a multilevel regulating system and by the balance of autonomic regulating processes.
Results and discussion. The analysis of seasonal variations in the composition of electrolyte metabolism of skiers revealed the following concentrations (by seasons: summer - fall - winter - spring): Calcium: 2,47±0,03; 2,27±0,01; 2,39±0,02; 2,31±0,01 mmol/l (control 2,25 - 3,00 mmol/l), Magnesium: 0,97±0,03; 0,92±0,01; 0,85±0,02 mmol/l (control 0,70 - 0,99 mmol/l), Potassium: 4,35±0,07; 4,15±0,04; 4,48±0,06; 4,14±0,05 mmol/l (control 3,48 - 5,30 mmol/l), Na: 140,64±0,99; 141,64±0,83; 140,48±1,22; 140,19±1,03 mmol/l, Fe: 151.2±0,6 mmol/l, Phosphorus: 1,12±0,07 mmol/l. The variations in the electrolyte composition were evidently seasonal; their dependence on intensity, type and orientation of trainings was revealed. The most stable was Na concentration. At upper levels of midland the Са2+ concentrations equaled 2,36±0,06 mmol/l, Mg2+ - 0,87±0,05 mmol/l, Potassium - 4,32±0,06 mmol/l, Sodium - 110,20±1,12 mmol/l, Phosphorus - 1,14±0,08 mmol/l, Fe - 15,47±0,87 mmol/l. The concentrations of bioelements for ski runners, participants of the Russian Federation championship (international masters of sports, masters of sports and candidate masters of sports), equaled: Са 2,35±0,02; Mg 0,91±0,01; Potassium 4,18±0,001; Sodium 142,12±0,06 mmol/l. As compared to experienced cross country skiers, young skiers had higher urea concentrations - 6,29±0,29 mmol/l. The real enzyme concentrations of cross country skiers were: AST 0,22±0,01; ALT 0,51±0,13 mmol/l (control соответственно 0,10-0,45 and 0,10-0,65 mmol/l respectively); AST, unit/l 10,30±0,39 (control 8-40); ALT, unit/l 24,12±2,14 (5-30); the AST/ALT relation (c.u.) 0,64±0,01 (0,80-1,20). The plasma protein concentration equaled 74,07±2,57 g/l (control 60-85 g/l); creatinine - 105,58±4,03 ^mol/l (control 55123). The urea concentrations varied in the range of 5,99±0,07 mmol/l (control 2,10-8,20 mmol/l).
Thus, seasonal variations in hemoglobin concentration were: 162,22±3,11 (summer); 150,21±2,95 (fall); 161,29±3,26 (winter); 145,64±2,52 (spring) g/l, in mountains 164,50±5,30 g/l. Thus, in the upper midland haemoglobin concentrations were higher, though in the ranges permitted by the WADA. The enzyme concentrations in lowlands varied with the seasons, respectively: 0,38±0,03 mmol/l; 0,39±0,04; 0,32±0,01; 0,33±0,02 (control 0,10 - 0,45 mmol/l), ALT: 1,10±0,09 mmol/l; 1,13±0,08; 1,09±0,09; 0,82±0,07 (control 0,10 - 0,68 mmol/l), AST: 17,53±0,32 unit/l, 17,89±0,97; 15,17±0,44; 15,79±0,50
(control 8±40 unit/l); ALT: 53,54±2,96 unit/l; 51,84±2,74; 52,95±2,87, 34,02±2,52 (control 5-30 unit/l).
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The AST/ALT relation varied as: 0,62±0,09; 0,60±0,08; 0,47±0,09; 0,58±0,08 c.u. In the mountains the ALT and AST equalled 44,33 ±2,59 and 48,50±4,14 units/l, respectively. The plasma protein concentration equaled 71,84±0,62 g/l, urea - 5,73±0,50 mmol/l, creatinine - 86,04±3,36 mmol/l. All these concentrations were in the reference range.
The urea concentration varied with the seasons as: 5,51±0,19 mmol/l; 5,31±0,12; 6,05±0,27; 4,77±0,16 (control 2,10-8,20 mmol/l), and in the mountains it was 7,48±0,42 mmol/l. The latter value evidences on intense efforts aimed at the acclimatisation, as well as big training loads (BTL) and fatigue.
The urea testosterone concentration also varied with the seasons: 14,25±0,52; 14,65±0,58; 12,67±0,41; 14,84±0,60, respectively; in the mountains it was 21,90±1,75 ^g per 8 l of blood. The acetylcholine concentrations were the following: 80,28±0,31; 80,51±0,30; 73,32±0,31; 78,66±0,22 ^g/ml (control 81,10 - 92,10).
The metabolic state of a 19-year old cross country skier (master of sports) prior to the start is given in Table 1.
Table 1. Clinical analysis of urine of a cross country skier (P. B., master of sports)
Parameters Results Normal range (control)
Fe 11.60 10-30 mmol/l
Mg 0.80 0.74-1.20 mmol/l
Ca 1/18 1.05-1.30 mmol/l
Creatine phosphokinase 118 25-200 units/l
AST 0.6 10-30 units/l
ALT 9.9 9-43 units/l
Urea 1.60 2.50-6.30 mmol/l
Testosterone 7.60 6.93 - 17.43 ^mol/l
Cartasol 154 N. 150-170 mmol/l
The alimentary deficiencies of certain microelements, particularly Fe and Zn, were associated with immunity reduction and increased susceptibility to infections.
About 50 % of Magnesium contained in human blood is in the bound state, and the rest is ionized. The Mg concentration in the human blood is 2,30-4,00 mg%. Mg complex compounds are delivered to the liver where they are utilized for the synthesis of bio-active compounds, along with urine and sweat. Magnesium is an intracellular element, it is involved in the metabolism, and it actively interacts with electrons. Magnesium also promotes the "bioenergetics" of essential processes, regulates neuromuscular massage, ocular muscle tone, stimulates protein formation and controls the ATP conservation and release. This element has an anti-stress effect, it reduces the excitation of nervous cells, strengthens the immune
system, has an antiarrhythmic effect, and increases recovery processes after the BTL. The Mg concentration in an athlete's body should be considered in combination with the bio-elemental homeostasis as a whole.
The Fe depletion or abundance is detrimental for the oxygen transport function, and it can block the intestine mucus from adsorbing other bioelements, such as Manganese [10]. Zn is a cofactor for many enzymes that are involved in protein and other types of metabolism. Zn depletion leads to the reduction of immune reaction on infectious agents.
Magnesium is neighbored in the periodic table by Calcium, and these elements are interchangeable in the human body, easily substituting each other in various compounds. Mg deficiency of a Ca-rich diet results in a delay in all tissues. Intense training activities accompanied by perspiration, sauna and high environment temperature lead to Mg losses.
The disorders in the metabolism of essential microelements can be a key cause of the secondary immune deficiency that can lead to reduced protein assimilability and caloric content even under the conditions of balanced intake of microelements. Upon deliberate weight losses, intensification of the T-cell immune response and inhibition of the antibody production can occur [5, 3, 9, 11].
Depending on age, sport, specialisation, body parameters and sport qualification, the protein intake should be discretely dosed at various stages of sport training [8]. This problem is complicated by the need for intense motor activity and the symbasic increased intake of proteins. There are a number of evidences on the physical activity promotion of heightened catabolism of proteins and accelerated withdrawal of the final products of the nitrogen metabolism from the body. The skiing causes changes in the concentrations of urea, creatine and uric acid in the blood and urine. Increased catabolism of proteins is observed in stress situations, including the conditions of high psychophysical stress, that evidences about the heightened demand of the body for proteins under intense muscular and psycho-emotional stresses.
Increased protein consumption is needed in sport where the muscular mass should be gained by young athletes. Continuous protein intake and growing physical activity cause a temporary increase in the positive nitrogen balance, with a high probability of subsequent development of hypoproteinemia and anemia. A severe and prolonged BTL leads to noticeable changes in the ratio of plasma protein fractions and in the protein nitrogen content.
Under the BTL conditions the activity of enzymes that are involved in the protein biosynthesis, and that increase the metabolism rate of muscle proteins, raises; the content of amino acids in the proteins of myofibrils, microsomes and mitochondria increases too [10]. The accelerated protein synthesis upon motor activity (MA) affects the formation of the adaptation mechanisms which increase the physical performance (PP) level and particular fitness. A diet with an increased protein intake promotes the protein synthesis in the muscles, and the catabolism is occurred to be inhibited. The MA-induced adaptation of
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functional and metabolic systems, including the enzyme pool and the hormonal regulation and integration spectrum, is significantly facilitated. The loss of body fat reserves and the muscle mass gain proceed along with intense input of proteins into the athlete's body. A.K. Euler [11] showed that "long-distance" athletes did not experience protein deficiency due to the following reasons:
- under the conditions of a long-term MA the synthesis of proteins in the connective tissues is suspended;
- during the BTL the proteins are insignificantly consumed for energy production;
Thus, the information about the protein metabolism of athletes is controversial. In some cases catabolism was observed to accelerate during muscular activity, whereas in others the synthesis of proteins was intensified after the BTL; some authors revealed acceleration of both catabolism and anabolic processes, others refuted any intensification of protein metabolism and its special role in the muscular activity [4,7].
The pharmacological means for maintenance and raise of the PP are described in the N.I. Volkov and V.N. Oleynikov's book [1]. One of the tasks of sport physiology is the correction of metabolic disorder, aimed at the PP heightening, adaptation reliability, immunologic resistance, acclimatization, and accelerated recovery of athletes after BTL damages.
The acceleration of natural post-exercise recovery can be provided by the natural detoxification means (detoxificants, antioxidants, rehydrants, hepatric remedies, sorbents, immunomodulators, vitamins, macro- and microelements, remedies for improving bloodstream through kidneys). Besides, the metabolism regulators, anabolic agents, psychomotor stimulators, sedative and nootropic drugs, neuroprotectors, correctors of blood microcirculation and rheological properties, blood production stimulators, and pH level regulators are applied. The ergogenic means are extremely important to be used in the training microcycles, considering current tasks. Among the components of energetic products (ATP, glucose, creatine, L-creatine), the metabolites of the three-carbon cycle acids take part in the muscle contractions. The energetic agents promote the recovery and the formation of an energetic depot, increase the glycogen resource and accelerate the transport of fatty acids from cytoplasm to mitochondria. The ATP, creatine phosphate and glucose are the energy sources in the anaerobic-aerobic zone. At a long-term BTL they activate the glycolysis [5, 2, 7].
Immune depression can be caused by the BTLs and biorhythm disordering which depress the immune system, indirectly influencing the AH and the susceptibility to infections. The hematologic homeostasis is maintained by the blood diluents which improve the microcirculation and rheological properties. The enterosorbents remove toxic substances from an athlete's body upon the BTL; those substances could have detrimental effect on the cardiopulmonary and immune systems, as well as on the blood production [10].
Distance-type, cyclic sports affect the cardiopulmonary system cumulatively through the neuromotor units. These sports need metabolic maintaining, particular functional diets, water balance maintenance and transfer from the hydrocarbon sources of energy (macroergic phosphates, glycogen, glucose) to lipids which play an important role in the correction of general state and the PP and in the control of hormonal state.
It should be noted that immunoglobulin (Jg) reacts on extreme environmental impacts in different ways. This is confirmed by the depression of the SCL of neutrophils and of the lysosomal activity of monocytes (factors of adaptation to stress caused by intense trainings and competitions). These functions are known to be directly related to the processes of severe affection of the parenchymatous tissues by stress impact. A comparison of the measurements carried out for young (17-18 year old) and adult (20-25 year old) judokas indicate the reduction of the lysosomal activity of neutrophils (Np) and of the NBT activity Mc and the constant Jg content during training process. Young athletes had heightened phagocytic Np and Mc activities, Np level induced by the chemiluminescence (ICL) and T-cells quantity. In the end of a microcycle, a valid decrease of Mc concentration, a raise in Mc lysosomal activity and a reduction of T-cell concentration and of ICL level of neutrophils were observed.
Thus, the adaptation improvement is provided by optimal receptor and metabolic interaction of the cell system. However, due to the BTL overloads a considerable fraction of athletes (1/4) belong to a "risk group" with the danger of development of secondary immune deficiency. Significant immune deficiency is observed for more than 30% of professional athletes. It is caused by chronic overloads during sport stresses, neuroimmune disorders, enzyme activity reduction, diminished protein content and misbalance between the humoral and cell components of immunity and the factors of non-special resistance, which determine the general biological basics of adaptation. All these factors are complemented by chrono-misadaptation and improper diets with protein and microelement (Fe, Zn, Mg, Se) deficiency.
The acute form of the disease of elite athletes during competition period 2,5-3,5 times increases as compared to the training period [2].
The integrative demonstrations of athletes' body functioning consist in the modification of the system that transfer from the simple ways of adaptation to complicated ones, changing structural and functional components at various adaptation phases.
The results of the immunological investigation of cross country skiers are listed in Table 2. Table 2. The immunological test sheet of cross country skiers, %
Cell type Normal range Actual values Normal range Actual values
Peripheral blood leukocytes 4.00-5.00 4.80
Segmented neutrophils 47.00-72.00 48.16
Eosinophils 0.50-5.00 2.70 0.02-0.30
Basophils 0.0-1.00 0.86 0.00-0.07
Monocytes 3.00-11.00 10.68 0.09-0.60
Peripheral blood lymphocytes 19.00-37.00 38.89 1.20-3.00 1.86
CO3+ cells (T-lymphocytes) 60.00-80.00 64.86 1.00-2.40 1.20
CO3+CO4+ cells (T-helpers) 33.00-50.00 45.50 0.60-1.70 0.63
CO3+CO8+ cells (cytotoxic lymphocytes) 16.00-39.00 17.16 0.30-1.00 0.32
CO 16+ cells (NV-cells) 3.00-20.00 18.69 0.03-0.50 0.35
CO 19+ cells (B-lymphocytes) 5.00-22.00 15.78 0.01-0.40 0.29
Immune regulation index (CO3+/ CO8+) 1.20-2.00 2.74
Phagocytal neutrophils (latex test) 55.00-95.00 86.20
The results of subpopulation analysis of lymphocytes were obtained from the data of cytofluometric measurements using an EPICSBECKMANCOULTER flow laser cytometer (USA) using the IMMUNITECH two-color monoclonal antibodies (France), the class of immunoglobulins was: JgM 1,18 mg/ml (0,10-2,30 mg/ml); JgA 2,64 mg/ml (0,80-2,50); Jg6 12,70 mg/ml (7,00-16,00 mg/ml); JgE 83,42 mU/ml (up to 87 mU/ml). The immunoglobulins in blood serum were detected by the immunoturbidimetric technique, JgE - by immunoenzymatic one. The immunoregulation index was heightened, and the relative values of helper cytotoxic T- lymphocytes were in the reference range. The concentration of A-class immunoglobulins in the blood serum was enhanced.
As seen from Table 2, the immunologic state of cross country skiers can be qualified as a stressed one. The majority of the components of immunological resistance were near the reference upper levels. The immunoregulation index exceeded normal values. The low values of the eosinophil index indicated the high activity of the neuroendocrine component (acetylcholine, glucocorticoids).
The analysis of wrestlers' peripheral blood at the annual stages revealed the amount of leukocytes in the range of 4,21-5,53x109 per liter, segmented neutrophils - 38,45 - 47,90%, eosinophils - 1,782,75%, monocytes - 5,80-7,20%, lymphocytes - 37,80-51,40%. The values varied with athletes' weight. The red blood parameters varied in the ranges: erythrocytes - 4,32-4,10x1012 per liter; hemoglobin -136,60-145,80 g/l.
The highest variations in the wrestlers' group were observed for the values of spontaneous chemiluminescence (SCL), 49,42-397.23 pulse/l; induced chemiluminescence (ICL), 29,54-140,56 c.u.; phagocytosis intensity, 85.18-274.53 c.u. The lysosomal activity of neutrophils (LAN) varied in the range of 311,23-696,91 c.u.; the intensity of monocyte phagocytosis changed from 74.78 to 286.93 c.u.; and the lysosomal activity varied from 93,24 up to 229,46 c.u..
The results of the multiple regression analysis proved the possibility to forecast the rank of sport mastership (RSM) and revealed systematic close correlations with increasing adaptation abilities of wrestlers; among the close correlations, the transformative non-linear relations between the RSM and regulators (muscle tonus component) and between the parameters and the RSM were observed. The parameters of biological processes, immunological resistance, including the peripheral blood cells, acted as protective adaptation effects. Adaptation covers various factors and regulation conditions, such as the activation of stress-realizing and stress-limiting systems under the conditions of structural and functional trace phenomena in the cell and humoral systems, cell membranes, biochemical processes (hormonal and enzyme activities), metabolism, myocardium, liver and kidneys, respiratory and neuromuscular systems. The adaptation (stress change) provides sparing of structural and functional resources, preventing the deterioration of organs, cells and systems, and modifying the interactions between special immune systems. An athlete's body activates new programs for energetic and protective processes during a long-term adaptation process. The direct and feedback connections of the RSM and immunoglobulins with parameters related to the athletes' state evidence about their direct effect on the sport efficiency. The adaptation of the wrestlers' functional systems under the conditions of circular and test training was considered. Physiological changes in the oxygen transport system and the acceleration of the post-stress recovery were revealed. A low concentration of the T-cells and normal values of the NBT (LAM activity) were observed. The key immunological parameters (SCL, NBT, LAM, LAN, NFN, NFM, T-cells, immunoglobins) varied in wide ranges. Prior to the circular and test training the initial immunological parameters of wrestlers had exceeded the control ones, whereas after the training they decreased to the donor levels. Some immunological components of several athletes were stressed, that caused their adaptation breakdown. Nevertheless, the investigation revealed that the variability of system-forming components in the reference range should be considered as an adaptive manifestation of stable homeostasis. The adaptation and stress factors have general direction that covers the key organic and systematic homeostasis under extreme environmental impacts, including periodic and chronic hypoxia.
The immunological reactivity is influenced by the pharmacological methods that impact the CNS, vegetotropy, hormones, antihistaminic remedies, cardiovascular compounds that affect the hemocoagulation, vitamins, antibiotics, hemiotherapy and anthelmintic medicines. The disorders in the immunological resistance at the BTL over the normal levels lead to the reduction of special working capacity. The so-called catarrhal deceases appear with increasing frequency, the athletes' immune state
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and resistance to various antigens are upset. The mesosomic activity is depressed under the long-term stressed conditions. On the contrary, the short-term intense stresses, such as wrestling throws, increase the lysosomal activity of neutrophils; the shock waves of training significantly depress the humoral and secretory components of the immune system. The phagocytic activity of leukocytes significantly decreased after the BTL, that was accompanied by reductions in the active phagocyte fraction, the phagocyte index, and the cell absorbing and digest ability. At moderate stresses the humoral factors of nonspecial resistance increased. A reduction of the A, 6, M immunoglobulin content in the blood serum after the BTL of 17-19 year old cross country skiers and wrestlers (sambo and judo) was registered [5]. After the BTL and competitions the T-cell immune system was depressed. The interrelations between the load intensity and the SCL T-cell depression level were revealed. The B-lymphocyte content after the BTL either did not noticeably change or increased, that caused the humoral immune heightening.
The JgM content in the athletes' blood serum did not change considerably during the preparation period, while raised at the single-stage loads. The Jg6 and JgA concentrations during the competition periods increased and somewhat reduced, respectively. The correlation between the concentrations of JgM and hydrocortisone (somatotropic hormone) in blood was revealed. The phase character of the changes in the humoral and cell components of the immune system, and the functional and the phagocyte activities of neutrophils and monocytes was observed. The remote from the BTL periods are least studied. However, the majority of authors report about the disorders of various immune functions upon the physical loads of high intensity according to the BTL system [6].
During the restitution, a secondary immune deficiency of the acquired clinical-and-physiological syndrome, consisting in the reduction of effector connections of the immune system and in the processes of nonspecial resistance, is observed. A risk of infectious deceases appears.
The immune system is damaged by the disorders in neuroendocrine, inner immune and neuromotor regulations, profound metabolic changes (heightened concentration of H+ and HC03- ions, reduced pH level, changed WE of 02 and W2 optimization, penetration of anomalous metabolites from highly stressed organs into the bloodstream). The deficiency of glucose, essential amino acids, polyunsaturated fatty acids, vitamins and microelements is also detrimental for the immune system. The main reasons of immune system weakening caused by adaptation for the middle attitudes are as follows:
• chrono-disadaptation caused by the flights to training and competitions sites;
• functional and metabolic states, special and physical working capacity;
• adaptation ability to the BTL, completeness and rate of recovery;
• over-training and overstrain, immune resistance level;
• diet quality, diet balance and functionality, sufficient amount of vitamins and microelements;
• availability or lack of the pharmacological correction of immune state of elite athletes.
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Conclusion. The intake of the preparation Essentiale combined with Levoton in pre-season prevents the decrease of the content of T-cells, their low variability at the phases of final pre-season in the black blood of judo and sambo wrestlers. Meanwhile, the content of B-lymphocytes was the lowest in the period of final pre-season, but in this period the content of T-cells, particularly B-lymphocytes, increased. The immunoglobulin concentration by the phases of the year cycles was changing in the following way: JgA - undulatory, exceeding the norm in the period of October and May competitions; Jg6 - over the reference limits in February-March, during the main duels; JgM -below the normal range in October, February and significantly - in March compared to the May data, when the period of major competitions was over. The decreased content of the production of T- and B-lymphocytes is related to the stress tension of the final training for big competitions (shock training sessions, wrestling days, qualification control combats). Changes in the Jg concentration depended on the content, direction, volume and manner of BTL.
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Corresponding author: [email protected]
