Phagocytic activity of leukocytes and blood cortisol ratings in athletes prior to and after phytopharmacological correction

PhD V.I. Vetosheva1
Dr. Biol., Professor V.V. Volodin2
PhD S.O. Volodina2
1Komi Science Center, Ural Branch of the Russian Academy of Sciences, Syktyvkar
2Institute of Biology, Komi Science Center, Ural Branch of the Russian Academy of Sciences, Syktyvkar

Keywords: phagocytic activity of leucocytes, lymphocytes, cortisol, phytopharmacological correction, Adasten, athletes.

Introduction. Long-term intense physical stresses in the training periods are known to activate the adrenal cortex and release the stress hormone cortisol into the bloodstream. It mobilizes the individual energy reserves to facilitate release of glucose and free fatty acids; and activates the processes of gluconeogenesis and protein catabolism [4]. As a result, the energy resource is replenished in the ATP synthesizing process to boost the performance. The catabolic effect of cortisol production, however, is of a negative effect on the lymphoid tissue as it suppresses lymphocytes in the bloodstream and phagocytosis to undermine the immunity [2].

To prevent adverse changes in the body, a method for formulation of the ecdysteroid-containing drug Serpisten from the top of the plumeless saw-wort (Serratula tinctoria) was developed. Being an adaptogen, as shown in animal experiments [1, 5], Serpisten can potentiate adaptive and regenerative processes and thus lead to the development of unspecifically increased resistance of the body [9]. The development of researches on the cellular mechanisms of influence of phytoecdysteroids on the human body is not just of obvious scientific interest but is also caused by the need to use such drugs in sports.

Objective of the study was to analyze benefits of ecdysteroid-containing dietary supplement Adasten by rating cortisol and the activity of blood system protection mechanisms in intense training conditions.

Methods and structure of the study. Sampled for the study were the Super League ice hockey players (n=20) aged 27±1.5 years with the 19±1.4 years of sport records on average. All of them were qualified Masters of Sport and Candidate Masters of Sport and were in the active training mode. The athletes were informed in detail on the ongoing experiment and gave their written consent to participate. The pharmacological support consisted in the administration of Adasten per os as a dose of 10 mg twice a day (20 mg daily) for 3 weeks. The Adasten administration course was used to obtain a delayed action with a gradual achievement of the supercompensation phase. We determined the total number of white blood cells (K/mcL) and the share of the formed elements of white blood (neutrophils, monocytes, and lymphocytes). The immunologic activity was assessed based on the rates of phagocytosis of latex particles by leucocytes, phagocytic index (FI) and NBT-test rates. The cortisol blood levels (nmol/l) were measured by the method of enzyme-linked immunosorbent assay using a set of reagents "Steroid-ISA-Cortisol". The ratio between the parasympathetic and sympathetic influences was determined by the Kerdo vegetative index. The studied parameters were compared before and after the Adasten administration course depending on the athletes' qualifications (MS and CMS) and age: 30- and 30+ year-olds [8].

Results and discussion. The initial cortisol levels in the examined athletes were on the average 578±37.7 nmol/l, with 21% of them tested with the above normal levels. The training loads increased the cortisol levels by 9.4% in the vast majority of athletes (72%), reaching 637±49.0 nmol/l (p>0.05), which exceeded the normal level in 39% of testees. This was due to the influence of physical loads and Adasten administration, which activated the hypothalamic-pituitary-adrenal and sympathoadrenal system [1]. The activation of the latter is evidenced by the fact that the Adasten administration course led to a pronounced shift towards the excitability of the sympathetic division of the autonomic nervous system. The cortisol levels after the phytopharmacological correction increased by 20% in the Masters of Sport of 31.6±1.4 years of age (p<0.05). The data obtained indicate that intense physical stresses activate the adrenal cortex, which is accompanied by increased secretion of glucocorticoids. Heightened environmental requirements led to adaptive shifts in the body of Masters of Sport through the mobilization of carbohydrates and fats as energy sources, while excessive and prolonged cortisol extraction under the influence of high physical stresses was accompanied by protein breakdown, which affected the lymphoid tissue, as a result of which the lymphocytes became an additional source of energy [4]. Inactivation of separate parts of the immune system can become a specific "cost" of adaptation, which makes it possible to predict the development of immune deficiency in these individuals [2, 4, 11].

The initial levels of lymphocytes in the group of athletes decreased by 16.5% (p<0.05). At the same time, after Adasten administration the lymphocyte levels in the blood of most athletes (80%), though decreasing in response to physical loads (p<0.5), remained within the normal limits (see Table). The comparative analysis revealed that for the 30- year-old athletes, most of whom were Candidate Master of Sport, the decrease was less pronounced (by 13%) (p<0.05), while for those over 30, most of whom were Masters of Sport, the decrease equaled 21% (p<0.05). The correlation analysis found no relationship between the blood levels of cortisol and lymphocytes prior to Adasten administration, while after phytopharmacological correction, an inversely proportional relationship (ρ=-0.4922; p<0.01) was detected during intensive trainings. It can be assumed that Adasten has a corrective effect on the blood lymphocytes, keeping them within the normal limits under physical stresses.

Table 1. Blood values of hockey players before (A) and after (B) phytopharmacological correction using Adasten

Value

Across the sample

30- year-olds

30+ year-olds

A

B

A

B

A

B

Cortisol, nmol/l

577.7

637.0

593.6

637.0

560.5

637.5

Lymphocytes, %

35.2*

29.4*

34.7

30.2

35.9*

28.5*

Leucocytes, K/mcL

5.4

5.2

5.7

5.7

5.1

4.7

Neutrophils, %

53.5

58.5

54.7

58.4

50.8

57.4

Monocytes, %

7.8

8.7

7.5

9.2

8.3

9.8

Phagocytic activity, %

66.3*

74.6*

62.0*

76.0*

69.6

73.7

Phagocytic index, %

12.0**

16.0**

11.7**

17.3**

12.2

15.0

NBT-test, %

33.3

28.6

25.4

31.2

34.0

26.7

________________

Note. The differences are significant at р<0.05 (*) and  р<0.01 (**).

Notations: NBТ-test – nitroblue tetrazolium test.

The total leukocyte count, both at the beginning of the training cycle and when performing physical loads against the background of Adasten administration, remained generally within the normal limits. However, 2/3 30+ year-old athletes were found to have a downward trend - by 7.5% (see Table 1). The initial monocyte levels in the group were also within the normal range, and for 21% - even higher. During Adasten administration, there was an upward trend in both age groups - by 15% and 18%, approaching the upper limit, and in 39% of athletes - above the norm. The same was true for neutrophils, and the changes were more pronounced for the 30+ year-old athletes. The performance of intense physical loads against the background of phytopharmacological correction kept the ratio between granulo- and agranulocytes in peripheral blood within the normal limits. This indicates the adaptogenic effect of Adasten, also known as ecdysteroid-containing preparations, both in animals [1, 7] and humans [3, 6, 10]. It should be noted that the decrease in the blood levels of lymphocytes under physical stresses against Adasten administration led to the increase in the neutrophil (ρ=-0.7416; p<0.01) and monocyte count, bringing the test items closer to the upper limit of normal. This indicated higher reactivity of cells and their ability to quickly engage in responses to adverse environmental factors, ensuring the maintenance of homeostasis. The high cortisol levels in athletes are known to reduce the production of phagocytes, which is reflected in the reduction of such phagocytic activity rates as phagocytic number and index of phagocytosis completeness. Phagocytosis is the first line of defense against infection, and the growth of cortisol rates can be considered as a risk factor for the development of infectious processes in athletes [2]. The NBT-test results showed an increase in the phagocytic activity of neutrophils and macrophages by 18.6% in the 30- year-old athletes and its decrease by 20.8% in the 30+ year-old individuals. Prior to Adasten administration, the NBT-test results showed that the norm was exceeded in 78.64% of individuals. Phytopharmacological correction led to the normalization of all indicators, suggesting the corrective effect of Adasten. For the group as a whole, the initial rates of phagocytic activity of leucocytes and phagocytic index were found close to the bottom of the norm (see Table 1). The pharmacological correction, despite the intensity of the athletes' training activity, increased the phagocytic activity of leucocytes and phagocytic index by 11.2% and 25.0%, respectively, indicating the biological activity of Adasten (p<0.05). A similar effect was observed during the experiments on the biological models [5]. The immunostimulating properties of adaptogens are particularly pronounced against the background of immune system suppression through activation of one of the most important components of non-specific resistance - phagocytosis [9]. For the younger athletes, the impact of Adasten on the phagocytic activity was more pronounced: the phagocytic activity of leucocytes and phagocytic index accounted for 18.5% and 32.4%, respectively. In the 30+ year-olds, the increase in these indicators was not significant (6% and 19%, respectively). The data obtained show that phagocytosis as a component of the immune response decreases with age.

Conclusions. The findings showed that during intense physical activity, the athletes' adrenal cortex is activated under the influence of Adasten, as a result of which the cortisol blood level is increased, which is more pronounced in the Masters of Sport. Thus, the autonomic nervous system is activated towards excitability. This enables the body to mobilize energy resources faster and better, allowing athletes to achieve better results. However, excessive excretion of cortisol that has a catabolic effect (in 39% of individuals it exceeded the norm) leads to the decrease in the number of lymphocytes in peripheral blood, which was greater in the group of Masters of Sport aged 31.6±1.4 years. However, due to the phytopharmacological correction during the training process, this indicator did not exceed the norm. The number of white blood cells remained the same, and the decrease in the lymphocyte count led to the increase in the levels of monocytes and neutrophils, bringing them closer to, and even exceeding, the upper normal limit of the norm. The detected redistribution of the cell pool indicates an increase in the cell reactivity and their ability to quickly engage in the response under intense physical stresses against the background of pharmacological correction. The phytopharmacological correction increased as well, reaching the upper limit of the norm, testifying to the biological activity of Adasten, with the effect more expressed in the 30- year-old athletes. This redistribution of the cell pool in favor of monocytes and neutrophils, increased activity of the blood protection system elements under intense physical stresses are indicative of the corrective effect of Adasten on immunity, being more pronounced in younger athletes. The identified peculiarities prove the expediency of Adasten administration by elite hockey players.

The study was carried out on the academic research topic, state registration No AAAA-A17-117121270025-1.

References

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Corresponding author: svetlana20664@yandex.ru

Abstract

Long-term intense physical stresses in the training periods are known to activate the adrenal cortex and release the stress hormone cortisol into the bloodstream. Cortisol mobilizes the individual energy reserves to facilitate release of glucose and free fatty acids; and activates the processes of gluconeogenesis and protein catabolism. As a result, the energy resource is replenished in the ATP synthesizing process to boost the performance. The catabolic effect of cortisol production, however, is of a negative effect on the lymphoid tissue as it suppresses lymphocytes in the bloodstream and phagocytosis to undermine the immunity. Objective of the study was to analyze benefits of ecdysteroid-containing dietary supplement Adasten by rating cortisol and the activity blood system protection mechanisms in intense training conditions. phytopharmacological correction of white blood cells was determined by latex phagocytosis rating method; and the cortisol in serum was rated by the enzyme-linked immunosorbent assay. Sampled for the study were the Super League ice hockey players (n=20) aged 27±1.5 years with the 19±1.4 year sport records on average.

The study found the cortisol rates growing over a training cycle in most of the sample, with the growth more expressed in the Master of Sports subsample; and the lymphocyte rates were found to faster drop in the 30+ years old players. The initial rates of the phagocytic activity of leukocytes were found close to the bottom of the norm. Intense physical stresses were found to increase phagocytic activity of leukocytes to the upper limit of the norm, with the effect more expressed in the 30- year-olds. Decrease in the number of lymphocytes with phagocytic activity of leukocytes growth was found indicative of redistribution of the cell pool in favor of monocytes and neutrophils that tend to grow. phytopharmacological correction using Adasten was tested to increase the activity blood protection system elements with good corrective effects and redistributing the cellular elements under physical stress, with the effect more expressed in the 30- year-olds.