Clinical studies were conducted to evaluate the effectiveness of the developed dietary supplement, based on vegetable oil and maral antler extract, containing phospho- and glycolipids. The dietary supplement was prescribed to a group of 30 patients (main group) for 4 weeks, once before or during meals in the first half of the day, in a dosage of 10 drops. The control group of patients did not receive any dietary supplements. Based on screening diagnostics, all patients were found to have autonomic nervous system (ANS) disorders. The patients in the main group had an average of 10±2 clinical manifestations of panic attacks (PA). A reliable decrease in clinical manifestations of PA was observed in all patients taking the dietary supplement (p<0.05). A complete alleviation of manifestations such as fear of death and the feeling of inability to control oneself at any given time was observed. There was a subjective improvement in well-being, manifested by decreased emotional instability, increased vitality and performance, increased work effectiveness, and decreased complaints about PA in all patients. The additional inclusion of nutritional supplements in the diet of patients with PA resulted in a significant redistribution of adaptive response types towards a reliable increase in the number of patients with a physiological response of training and activation, and a reliable (p < 0.05) decrease in the number of patients with acute and chronic stress. Based on the studies, it was concluded that the additional inclusion of dietary supplements in the diet leads to reliable normalization of the functional state of the autonomic nervous system, increased overall performance, and adaptive responses. A positive dynamic was observed in the state of the central and autonomic nervous system. The mechanism of ANS harmonization involves a decrease in tone via the sympathetic nervous system, followed by normalization under the influence of the parasympathetic nervous system.
Introduction
The modern period of development in human society is accompanied by significant changes in the neurophysiology of the organism. This is due to the rapid development of civilization and its consequences:
Particular attention is paid to the impact of these factors on the growth of functional disorders of the autonomic nervous system (ANS), including those that manifest as panic attacks (PA) (Goodkind et al., 2020; Park et al., 2020; Abieva, 2021; Amakacheva & Morozova, 2021; Sander et al., 2022; Kennedy et al., 2023; Garbarova & Vartiak, 2024).
A panic attack or vegetative crisis is an extreme form of vegetative-vascular dystonia, characterized by symptoms such as panic disorders, nervousness, a feeling of unreasonable fear, and anxiety (Evstratova & Gorbunova, 2020; Kalchenko, 2020; Kanana & Baeshko, 2020; Abieva, 2021; Amakacheva & Morozova, 2021; Gurbanova, 2021; Alanazi et al., 2022; Efremov, 2023).
The term PA is officially included in the International Classification of Diseases, 10th revision (ICD-10), under the code F43.3. Approximately 40% of individuals who initially seek medical help from general practitioners exhibit symptoms of PA. The main age group affected is working-age patients, underscoring the importance of addressing this issue and developing solutions (Chereshnev & Poznyakovsky, 2020; Elias et al., 2020; Karthikeyan et al., 2020; Abieva, 2021; Sergun et al., 2022; Subramani & Sipkema, 2022; Ganea et al., 2023; Mikhailovich et al., 2023; Komov et al., 2024; Poznyakovsky, 2024).
The optimal approach to managing PA attacks is considered to be a scientifically-based combination of drug and non-drug therapy. Non-drug therapy should include the use of adaptogens, a pharmacological group of naturally occurring substances, such as maral antlers. These adaptogens have the ability to enhance the body's non-specific resistance to a wide range of harmful effects from chemical, physical, and biological sources (Menni et al., 2020; Musial et al., 2020; Sarris et al., 2020; Davinelli et al., 2021; Lopresti et al., 2021; Jiang et al., 2021; Scholey et al., 2021; Li et al., 2022; Siddiqi et al., 2022; Wu et al., 2022; Sultangazy & Tilek, 2023; Theivasigamani & Palaniappan, 2023; Tsvetkova et al., 2023; Bakhuraysah et al., 2024; García et al., 2024; Kim et al., 2024; Zhang et al., 2025).
Materials and Methods
Tests were conducted on a new dietary supplement preparation based on vegetable oil and lipids (phospho- and glycolipids) extracted from the antlers of the Altai maral.
Two groups of patients were selected: the main group and the control group, each comprising 30 patients aged 18 to 55 years.
The dietary supplement was administered orally sublingually, with 10 drops once before or during meals in the first half of the day for 4 weeks. The control group did not receive the dietary supplement.
Immediately after the diet therapy course ended, clinical studies were conducted to assess effectiveness. A 2-week observation period followed to monitor the relapse of panic attack episodes.
The state of vegetative tone was determined by quantitatively assessing the presence of vegetative dystonia syndrome (Vain AM, 2003. "Research schemes for identifying signs of vegetative disorders").
The clinical effectiveness of the dietary supplement was evaluated by analyzing the main symptoms of PA using a questionnaire developed for this purpose. This assessment was done before and after 4 weeks of taking the specialized product. The diagnosis of "Vegetative-vascular dystonia of the PA type" was confirmed in patients who exhibited at least 4 symptoms.
The type of adaptation reaction was determined by the percentage of lymphocytes in the leukocyte formula and their ratio to segmented neutrophils using the method developed by E.D. Golberg and colleagues in 1996.
Heart rate variability (HRV) was studied by recording short fragments of a standard cardiointervalography record using a hardware and software complex. The following parameters were assessed during the cardiointervalography analysis: stress index, heart rate, variation range (characterizing the degree of influence of the parasympathetic division of the autonomic nervous system on the heart rate), mode (reflecting the state of the humoral channel of heart rate regulation), integral indicator of the level of functioning of the central circuit of heart rate regulation.
Rate regulation (indicating the degree of centralization of control of the heart, taking into account the state of the humoral channel of regulation and the ratio of the activity of the parasympathetic and sympathetic divisions of the ANS), and mode amplitude (showing the degree of influence of the sympathetic division of the ANS on the heart rate).
The levels of the main stress-resistant cytokines IFNγ, IL-2, IL-4, and IL1β in the supernatants were determined using the solid-phase enzyme immunoassay "sandwich" method, adsorbed on the solid phase of a microplate (the procedure was carried out according to the instructions of "Procon" (Russia) and "Cytimmune" (USA).
The studies were conducted at the scientific and clinical center for hormonal health "ProfMed" in the Department of Obstetrics and Gynecology at the Siberian State Medical University (Tomsk) under the supervision of Doctor of Medical Sciences, Professor L.S. Sotnikova.
Results and Discussion
During screening diagnostics, pronounced disorders of the autonomic nervous system were observed in all patients. The total score in the main group with panic attacks did not align with the standard indicators (below 10 points) and averaged 85.3±5.05, exceeding the norm by more than 8 times. Patients in the main group exhibited an average of 10±2 clinical manifestations of panic attacks (Alanazi et al., 2022; Salikhova et al., 2023; Česaitis et al., 2024).
Table 1 displays the results of the study on the number of patients who experienced clinical manifestations of panic attacks before and after taking the dietary supplement (Juhari et al., 2023; Zhang et al., 2023; Cakmak et al., 2024).
Analysis of changes in vegetative-vascular dystonia of the panic attack type revealed a significant (p< 0.05) reduction in clinical manifestations of panic attacks in all patients taking the dietary supplement. It is noteworthy that symptoms such as fear of death and the feeling of losing control were completely alleviated (Saeed, 2022; Bei et al., 2023; Elamin et al., 2023; Rajopadhye et al., 2023).
No complaints related to the dietary supplements were reported, and no side effects were noted. Patients subjectively reported improved well-being manifested by reduced emotional volatility, increased vitality, and enhanced work performance. Complaints related to panic attacks decreased in all patients (Donati et al., 2022; Domatskiy & Sivkova, 2023; Hambețiu et al., 2023; Getu et al., 2024; Leadbeatter & Tjaya, 2024).
Table 1. Clinical manifestations of panic attacks before and after taking the specialized product (number of patients experiencing this manifestation)
|
Clinical manifestations of panic attacks |
Dietary supplement before treatment n =30 |
Dietary supplement 4 weeks reception n =30 |
Control group n =30 |
|
Internal trembling, trembling in the hands, chills |
28 |
5* |
0 |
|
Increased sweating, cold, clammy sweating |
14 |
3* |
0 |
|
Increased pulse, rapid, and increased heartbeat |
22 |
6* |
0 |
|
Abdominal discomfort, nausea |
19 |
8* |
0 |
|
Pain or discomfort in the heart area |
28 |
10* |
0 |
|
Difficulty breathing, feeling of suffocation |
19 |
4* |
0 |
|
Fear of doing something, going crazy |
12 |
2* |
0 |
|
Perception of the surrounding world as distant and not real |
10 |
2* |
0 |
|
Perception of oneself as if from the outside, a feeling that you are not able to control yourself at the moment |
26 |
0* |
0 |
|
Dizziness, feeling of loss of balance, loss of consciousness |
24 |
3* |
0 |
|
Inability to sleep |
30 |
8* |
0 |
|
Unpleasant sensations in the hands and feet: coldness, tingling, numbness |
22 |
4* |
0 |
|
Confusion of thoughts, inability to concentrate |
30 |
5* |
0 |
|
Fear of dying |
28 |
0* |
0 |
Note: * - significant differences (p< 0.05) compared to "before treatment".
A thorough assessment of the functional state of the adaptive potential of the autonomic nervous system was conducted (Table 2).
Table 2. The distribution of adaptation reactions in patients before and after taking the specialized product (X ± m, p)
|
Type of adaptive response |
Dietary supplements before treatment n=30 |
Dietary supplement intake 4 weeks n =30 |
Control group n=30 |
|
Training and activation reactions |
0 |
18* |
30 |
|
Acute stress |
14 |
2* |
0 |
|
Chronic stress |
16 |
10* |
0 |
Note: * - significant differences (p< 0.05) compared to "before treatment".
The use of dietary supplements in patients with PA resulted in a significant shift in the types of adaptive reactions, leading to a notable increase in the number of patients displaying a physiological reaction of training and activation, as well as a significant (p< 0.05) decrease in the number of patients experiencing acute and chronic stress.
The Kerdo vegetative index in patients averaged 44.2±2.32 arbitrary units, indicating a clear predominance of stimulating influences on the activity of the autonomic nervous system. The dominance of sympathetic tone leads to tension in the body's adaptive capabilities, clinically manifested by symptoms of autonomic irritation and episodes of panic attacks as revealed during the collection of patient history and questionnaire surveys (García & Jaramillo, 2023; Varoneckaitė et al., 2024).
When assessing heart rhythm parameters based on cardiointervalography results, all patients showed increased sympathetic ANS tone and decreased parasympathetic regulation. This was evidenced by a statistically significant increase in mode amplitude, tension index, and variation range compared to normal values. The highest sympathicotonia was observed in the 35-45 age group. These findings suggest an imbalance in ANS components toward sympathicotonia, centralization of regulatory processes, and a decrease in adaptive mechanisms (Sugimori et al., 2022; Poornachitra & Maheswari, 2023; Samaranayake et al., 2024).
It was concluded that dietary supplementation reliably normalizes the autonomic nervous system's functional state, enhancing overall performance and adaptive responses. Positive changes in psychopathological symptoms were observed through psychometric assessments, showing a normalization of the psycho-emotional background, and a significant decrease in patient asthenization. Half of the patients reached a satisfactory functional state score of 8-10 points. Reactivity of the sympathetic ANS division during the orthostatic test significantly decreased, while parasympathetic activation was adequate during functional tests. The harmonization mechanism reduces sympathetic tone and normalizes it through the parasympathetic system (Pisano et al., 2023).
Table 3 illustrates the impact of nutritional factors on the functional state of the autonomic nervous system.
Table 3. Indicators showing the autonomic nervous system state in patients before and after taking the specialized product (X ± m, p)
|
ANS indicator |
Dietary supplements before treatment n=30 |
Dietary supplement intake 4 weeks n =30 |
Control group n=30 |
|
Kerdo index, arbitrary units |
44.24±2.32 |
12.28±0.02* |
2.23±0.04 |
|
Mode, sec. |
0.80±0.13 |
1.98±0.12* |
2.46±0.27 |
|
Mode amplitude, % |
78.27±3.01 |
40.10±1.25* |
38.14±2.11 |
|
Variation range, sec. |
1.10±0.05 |
0.22±0.04* |
0.28±0.02 |
|
Tension index, units |
223.24±4.08 |
124.35±2.05 |
138.23±5.12 |
Note: * - significant differences (p< 0.05) compared to "before treatment".
The study of heart rate spectrum indices, as one of the options for assessing the state of vegetative support of functions, was conducted 4 weeks after the start of the dietary supplement prescription. According to literature data, such a period is quite sufficient to stop vegetative reactions in the form of panic attacks, which is confirmed by the correction of vegetative dysfunction in the conditions of diet therapy.
A comprehensive analysis of hormonal status indicators revealed statistically significant (p<0.05) trends toward normalization of stimulated insulin and cortisol levels with the dietary supplement (p<0.05). No changes were found in thyroid hormone and prolactin levels.
Normalization of the stimulated insulin level is a consequence of cortisol normalization, achieved through a specialized product with adaptogenic properties that neutralizes the pathological influence of the sympathetic nervous system. As a result, cortisol levels and, consequently, insulin are normalized, as evidenced by the activation of the insulin-stimulating effect, indicating a positive effect of the nutritional factor on stress-dependent hormones (Table 4).
Table 4. Hormonal status in patients before and after taking the specialized product (X ± m, p)
|
Index (SI units) |
Dietary supplements before treatment n=30 |
Dietary supplement intake 4 weeks n =30 |
Norm |
|
Basal insulin (microunits/ml) |
12 [ 8:14] |
10 [ 8:16] |
2.7 – 10.4 |
|
Stimulated insulin (microunits /ml) |
32 [ 22:40] |
14* [ 8:18] |
2.7 – 10.4 |
|
Cortisol (nmol /l) |
840 |
550* |
150 - 650 |
|
Prolactin (mU /l) |
590 [ 500:600] |
535 [ 420:550] |
109 – 557 |
|
TSH (mU /l) |
3.6 [ 2.6:3.8] |
3.4 [ 2.8:3.6] |
0.4 – 4.0 |
Note: * - significant differences (p< 0.05) compared to "before treatment".
The association between the biological correlation of stress response and somatic complaints before the onset of clinically expressed psychopathological disorders, such as panic attacks, was studied. Early predictors of vulnerability to somatization include increased blood cortisol levels and decreased heart rate variability. These changes significantly reduce the body's resistance to the effects of typical psychosocial stressors.
The relationship between manifestations of the mental sphere and the nervous, endocrine, and immune systems has been established, forming the basis for the functioning of the psychoneuroimmunoendocrine system. Almost all immune cells have receptors for hormones associated with the sympathetic and adrenal axes, known as stress hormones. Immune modulation by these hormones can occur directly by binding to their receptors on the cell surface or indirectly by dysregulating cytokine production, which affects the metabolism of key neuropeptides and neurotransmitters (such as tryptophan, serotonin, norepinephrine, dopamine, and GABA). The balance between cytokines plays a crucial role in modulating cellular reactions in the brain during psychological stress and mental disorders, including panic attacks.
Cytokines serve as intercellular signaling factors and regulatory factors for not only the immune system but also the hormonal and nervous systems. The balance of cytokines significantly impacts psycho-emotional status. Evaluating the effect of the studied form of dietary supplement on the cytokine profile is essential for understanding the mechanisms of its action in panic attacks.
Table 5 presents the results of spontaneous and FGA-stimulated production of Thl and Th2 lymphocytes in peripheral blood.
Table 5. Spontaneous and FΓA-stimulated production of Th1 and Th2 cytokines by peripheral blood lymphocytes (X ± m, p)
|
Registered Indicator |
Surveyed Groups |
|||
|
Dietary supplements before treatment n=30 |
Dietary supplement intake 4 weeks n =30 |
Control group n=30 |
||
|
IL-1β |
Spontaneous, pg /ml |
12128± 6,87 |
79.97 ± 532 (p< 0.05) |
50.91 ± 3.09 |
|
FGA-stimulated, pg /ml |
95.46 ± 532 |
103.46 ± 3.09 |
132.87 ± 5.97 |
|
|
Stimulation index |
2.25 ± 0.02 |
2.29 ± 0.03 |
238 ± 0.05 |
|
|
IL- 2 |
Spontaneous, pg /ml |
2325 ± 3.91 |
52.02 ± 1.15 (p< 0.05) |
46.70 ± 3.65 |
|
FGA-stimulated, pg /ml |
105.63 ± 4.02 |
152.09 ± 5.02 (p< 0.05) |
19630 ± 8.42 |
|
|
Stimulation index |
1.98 ± 0.02 |
5.52 ± 0.03 (p< 0.05) |
438 ± 0.08 |
|
|
IL - 4 |
Spontaneous, pg /ml |
11633 ± 5.92 |
7933 ± 3.79 (p< 0.05) |
58.89 ± 334 |
|
FGA-stimulated, pg /ml |
23631 ± 11.02 |
193.23 ± 7.02 (p< 0.05) |
146.05 ± 624 |
|
|
Stimulation index |
2.03 ± 0.02 |
2.46 ± 0.03 (p< 0.05) |
2.48 ± 0.03 |
|
|
IFNγ |
Spontaneous, pg /ml |
116.82 ± 7.09 |
179.82 ± 5.15 (p< 0.05) |
206.01 ± 5.78 |
|
FGA-stimulated, pg /ml |
123.09 ± 5.57 |
207.42 ± 624 (p< 0.05) |
25632 ± 8.09 |
|
|
Stimulation index |
0.97 ± 0.02 |
1.14 ± 0.03 |
2.61 ± 0.02 |
|
Note: significant differences (p< 0.05) compared to "before treatment".
Activation of Th1 lymphocytes, along with the production of key cytokines IL-1β, IL-2, and IFNγ, enhances cellular immunity. The determination of the immune response via the humoral pathway is predominantly influenced by Th2 cytokines, with IL-4 serving as the discriminator. Study of the cytokine-producing function of blood mononuclear cells in the PA group revealed changes in parameters characterizing the ability of immunocompetent cells to secrete cytokines.
The study showed a significant increase in spontaneous mononuclear IL-1β secretion in patients with PA, which decreased after dietary supplement use to levels similar to those of the control group. The PHA-stimulated production of the interleukin and the stimulation index were consistent across all groups.
All PA subjects exhibited suppression of both spontaneous and PHA-stimulated production of IL-2, a determinant immune response in the Th1 pathway implementation. The cytokine secretion index in this group was notably lower than in the control groups. Diet therapy helped restore IL-2 cytokine production, bringing levels closer to control values. There was a significant increase in both basal and PHA-stimulated production of IL-4 by mononuclear cells, promoting the polarization of T-helpers towards the Th2 pathway. Even after discontinuing the dietary supplement, cytokine secretion by immunocompetent cells remained significantly higher than control values.
It was discovered that both spontaneous and induced production of IFNγ by immunocompetent cells was lower than the parameters of the control group, indicating a lower level of antiviral immune protection in PA patients. The use of dietary supplements helped restore IFNγ synthesis by immunocompetent cells, although it remained lower than the level in the control group.
Conclusion
It was concluded that the nutritional factor contributes to the normalization of spontaneous and PHA-stimulated cytokine production, including IFNγ, IL-2, IL-4, and IL1β. These cytokines play a crucial role in the joint functioning of the nervous and immune systems, which ensures stress resistance and adaptation of the body.
Following the completion of the diet therapy course, patients were observed for two weeks to evaluate the recurrence of PA episodes. Out of 30 patients, 12 experienced PA, but the severity of their episodes was significantly reduced compared to before the dietary supplement was used. The duration of PA episodes ranged from 10 to 20 minutes, with an average of 10±5 minutes. The frequency of attacks was also notably reduced post-diet therapy. Therefore, the results of the studies suggest that the dietary supplement has several mechanisms of action when used in courses for patients with PA:
The combined effect of these mechanisms leads to a reliable reduction in the severity, duration, and frequency of PA episodes.
Analysis of the clinical trial results for the developed dietary supplement indicates that it is effective for preventing and treating vegetative-vascular dystonia of the PA type.
Acknowledgments: The authors thank the administration of Research and Production Association «Biolit» for the opportunity to research its basis.
Conflict of interest: None
Financial support: None
Ethics statement: The study was conducted in accordance with the guidelines of the Declaration of Helsinki at the Scientific and Clinical Center for Hormonal Health "ProfMed" within the Department of Obstetrics and Gynecology at Siberian State Medical University (Tomsk) and the Testing Protocol No. ZF000001145 dated March 25, 2025.
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