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EFFECT OF PUNCTURE PHOTOBIOMODULATION ON PAIN STRESS: A PLACEBO-CONTROLLED EXPERIMENTAL STUDY IN RATS
A.G. Polyakova1, A.G. Soloveva2, P.V. Peretyagin2*, A.M. Rezenova1
1 Federal State Budgetary Educational Institution of Higher Education «Privolzhsky Research Medical University» of the Ministry of Health of the Russian Federation, 10/1 Minin and Pozharsky Square, Nizhny Novgorod, 603005, Russia;
2 National Research Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Ave., Nizhny Novgorod, 603022, Russia.
* Corresponding author: [email protected]
Abstract. The aim: determination of correlation dependence of adaptogenic defense reactions in rats in the acute phase of pain stress on laser-puncture exposure to low-intensity IR radiation. Material and methods. Four groups of rats were used in the work: "experimental", which received after the injury a 10-day course of PPBM, and three comparison groups ("intact", "no exposure" and "placebo"). The intensity of lipid peroxidation, specific activity of antioxidant enzymes, protein concentration, malonic dialdehyde content in blood plasma and erythrocytes were determined by spec-trophotometric method. Microcirculation indices were evaluated according to the previous series of our studies. Emotional and behavioral reactions were studied in the "open field" test. Results. Normalization of metabolic, microcircu-latory and behavioral indices was registered in the experimental group. The correlation coefficients between the microcirculation index and the data of behavioral activity under pain stress were significantly less than the correlation coefficients between oxidative stress and cognitive functions, which confirms the determining role of oxidative metabolism disturbance in the formation of cognitive disorders. Conclusion: the registered adaptation and stress-limiting effect of PPBM in conditions of experimental pain stress allows us to recommend the technology of laser acupuncture for use in complex rehabilitation of patients with pain syndrome.
Keywords: experimental stress, pain syndrome, oxidative metabolism, microcirculation, adaptive behavioral reactions, laser acupuncture.
List of Abbreviations
AP - acupuncture points DC - diene conjugates IR - infrared
LDF - laser Doppler flowmetry LPEC - latent period of exit to the center LPFM - latent period of first movement LPO - lipid peroxidation MDA er - malonic dialdehyde in erythrocytes
MDA pl - malonic dialdehyde in blood plasma
NCS - number of crossed squares
OF - open field
OS - oxidative stress
PPBM - photobiomodulation
SB - Schiff bases
SOD - superoxide dismutase
TC - triene conjugates
Introduction
The fight against pain stress is an urgent problem of modern medicine. The body's sys-
temic response to stress, aimed at eliminating or weakening it, is accompanied by the development of adaptive and compensatory reactions, including metabolic, vascular, autonomic and behavioral responses. The body's universal adaptive response to severe injuries and diseases of humans and animals, accompanied by pain, is oxidative stress (OS), which is expressed by an imbalance between the body's anti- and prooxidant defenses (Kelley et al., 2017; Olefir et al., 2021). At the same time, unfavorable stress-induced emotional reactions occur, manifested by increased excitability, fear and anxiety. Correction of these manifestations using drug therapy and classical acupuncture is fraught with undesirable side effects and complications (Al'-Zamil', 2020; Ponomarenko, 2022), which made the method of physiopunc-ture promising: a combination of exposure to acupuncture points (AP) with specific physical agents, which summarizes their capabilities. Thus, the action of punctuated photobiomodulation (PPBM) is largely based on the homeo-
static mechanisms of laser radiation, and at the same time, retains the advantages of punctuated reflex effects (minimum contraindications, low energy dosages, safety and economic feasibility), which is due to the histomorphological, biophysical features of AP, their connection with the autonomic nervous system being confirmed by clinical and experimental studies (Berdni-kova, 2019; Al'tman & Al'tman, 2019; Gur'janova, 2008; Chon etal., 2019; Jelinkova,
2013).
The monograph (Wattanatorn & Sutalangka,
2014) presents data from an experimental study of PPBM (so-called laser puncture) on laboratory rats to relieve acute inflammatory, visceral and neuropathic pain. From the standpoint of evidence-based medicine, the positive effect of laser acupuncture at the HT7 point on cognitive deficits, neuronal loss, oxidative stress, and the functions of the cholinergic and dopaminergic systems in a model of Parkinson's disease has been confirmed (Aleksandrovskaja & Kruglova, 2021).
However, the lack of analysis of the correlative dependence of corrective effects and a broad evidence base on the possible development of undesirable effects hinders the widespread introduction of PPBM into rehabilitation protocols of patients with consequences of injuries and diseases complicated by pain syndromes. This encourages the continuation of experimental work aimed at controlling antiox-idant and cognitive adaptation reactions in animals under conditions of pain stress, identifying the degree of correlation between impaired oxidative metabolism and the development of emotional-cognitive disorders.
Aim of the study: to determine the correlation between adaptogenic metabolic and stress-protective reactions during low-intensity laser puncture in rats in the acute phase of experimental pain stress.
Materials and Methods
We carried out a placebo-controlled experimental study of the correlation dependence of indicators of oxidative metabolism, motor and emotional-exploratory activity in 40 male Wistar rats weighing 250-300 g. in the acute
STRESS: S
stage of pain stress against the background of PPBM. The work used the minimum permissible number of laboratory animals in accordance with the applied method of statistical processing of the results obtained. The rats were kept in standard vivarium conditions: natural light, a balanced diet and free drinking. The pain syndrome model was induced by double li-gation of the sciatic nerve to the site of its bifurcation under intramuscular anesthesia (Zoletil + Xylazine) (Moskvin & Agasarov, 2019). The study was carried out in strict accordance with the ethical standards and rules of laboratory practice of the Geneva Convention for the Protection of Animals "International Guiding Principles for Biomedical Research Involving Animals" (Geneva, 1990), order of the Ministry of Health of the Russian Federation dated May 18, 2021 N 464 n "On approval of the Rules for conducting laboratory research" (with amendments and additions) (Peretyagin et al, 2011) and approval of the Local Ethics Committee of the Federal State Budgetary Educational Institution of Higher Education "PRMU" of the Ministry of Health of Russia (protocol No. 6 of April 29, 2022).
The animals were divided into four groups: an experimental group and three control groups (10 animals in each). Rats of the experimental group in a state of pain stress received a course of contact exposure to a laser irradiator of a certified device "Spectrum LC" (Russia) in the near infrared (IR) range on AP GV.14, localized in the area of the occipital protuberance, at the skin projection of the center of autonomic regulation responsible for development of adaptive reactions of the body. Additionally, the BL.37 point, located in the area of sciatic nerve ligation, was affected. Energy parameters of the effect: wavelength (X) - 845 ± 35 nm, average radiation power (P) - 0.35 ± 7 mW (exposed in mode P = 50% of the average), procedure time - 10 minutes (Kuz'mina et al., 1993). Irradiation sessions began immediately after the cessation of surgery and continued daily for 10 days. During the procedure, the rats were fixed in a special pencil case.
The control groups were comparison groups: intact rats (control 1), a group of animals with
pain stress "without exposure" (control 2) and the placebo group (control 3), in which imitation irradiation was used. The animals were removed from the experiment at the end by decapitation under anesthesia (Zoletil + Xylazine). To assess oxidative metabolism parameters, blood stabilized with sodium citrate (1:9) was used. The intensity of lipid peroxidation (LPO) was assessed by the concentration of malonic dialdehyde, diene conjugates (DC), triene conjugates (TC) and Schiff bases (SB) (Mihara et al., 1980), the specific activity of antioxidant enzymes - catalase (Sirota, 2016) and superoxide dismutase (SOD) (Sibgatullina & Haertdinova, 2011). Protein concentration was calculated using the Lowry method modified (Hall, 1936). The content of malonic dialde-hyde in blood plasma (MDA pl) and erythro-cytes (MDA er) was determined by reaction with thiobarbituric acid using TBA-AGAT reagent kits (Agat-Med LLC, Russia) on a PE-5400 spectrophotometer (Russia).
The dynamics of emotional-behavioral adaptive reactions were studied according to data from indicators of motor and emotional-exploratory activity in the "open field" (OF) test developed by K. Hall (Majorov, 2011). The OF test is considered a classic informative, simple and often used in world practice to assess the motor and orientation-exploratory activity of animals (Sudakov et al., 2013). The analysis of the results was carried out based on horizontal and vertical motor activity, grooming (washing), assessed as an element and an additional index of orienting-exploratory motivation, since grooming is very sensitive to the level of anxiety (Savin et al., 2017). To assess the dynamics of skin microcirculation, the laser Dop-pler flowmetry (LDF) method was used. The laser analyzer "LAKK-M" (NPP "Lazma", Russia) was used. During the study, the analyzer probe was installed perpendicular to the area under study. The recording duration was 3 minutes. We assessed the integral microcirculation index, which characterizes the degree of perfusion of tissue volume per unit time, the role of active ones (endothelial oscillations - 0.01-0.08 Hz, neurogenic oscillations - 0.080.2 Hz, myogenic oscillations - 0.2-0 .7 Hz)
and passive (respiratory - 0.7-2 Hz, cardiac -2-5 Hz) factors regulating microcirculatory blood flow with further calculation of the bypass index (Bajrovic et al., 2000; Krupatkin & Sidorov, 2013).
Statistical data processing was carried out using the Statistica 6.0 program (Stat Soft, Inc.). To test the hypothesis that the distribution of the obtained variants corresponded to the normal distribution, the Shapiro-Wilk test was used. Data are presented as M ± o. Student's t-test was used to assess the statistical significance of differences. When calculating the Student's t test, a Bonferroni correction was used to allow for multiple comparisons. The critical value of the significance level is taken to be 0.05. To analyze conjugate indicators, the Pearson correlation coefficient (r) was calculated (with normal data distribution).
Results
A study of indicators of oxidative metabolism revealed in animals in a state of painful trauma (control 2) the development of OS, characterized by an increase in the concentration of lipid peroxidation products (LPO) against the background of a decrease in antioxidant enzymatic activity. The content of primary LPO products - diene conjugates - in animals of the "control 2" group increased by 13% (p < 0.001), the concentration of secondary LPO products -triene conjugates (in plasma) and MDA (in erythrocytes) - increased by 17% (p < 0.001) and 44% (p < 0.001), respectively, the level of lipid peroxidation end products (SB) increased by 17% (p = 0.002) compared to the indicators of intact rats. Since the highly toxic primary, secondary and final products of lipid peroxida-tion (DC, TC and SB) are distinguished by their ability to damage proteins, lipoproteins and nucleic acids, their growth during OS leads to de-stabilization of membranes and cell degradation. The specific activity of the antioxidant enzymes SOD and catalase decreased in erythrocytes by 8% (p = 0.024) and 10% (p < 0.001), respectively, compared with the indicators of the group of intact rats (Table 1). Thus, acute pain stress in animals was accompanied by inhibition of antioxidant protection of the blood,
an increase in free radical processes with the accumulation of lipid peroxidation products both in plasma and in red blood cells.
In the experimental group after irradiation during pain stress, compared with the "control 2" group, a statistically significant increase in the specific activity of SOD by 18% (p < 0.001) was revealed. There was also a decrease in the concentration of the studied lipid peroxidation products in plasma and red blood cells compared to the "control 2" group: DC - by 5% (p = 0.026), MDA plasma - by 8% (p < 0.001), MDA erythrocytes - by 7% (p < 0.001), TC -by 8% (p < 0.001), SB - by 9% (p < 0.001). At the same time, in the placebo group (control 3), similar indicators remained close in value to the group "without exposure" (control 2), which reliably confirms the effectiveness of laser puncture in experimental OS.
The analysis of behavioral data in the "open field" test was carried out according to the orient-ing-exploratory reaction (the number of stances and horizontal motor activity, the stability of nervous processes reflected in the number of reverse turns, the time of visiting the central brightly lit squares), as well as according to the vegetative and emotional reaction (according to grooming time). The results of the behavior of rats (motor and orientation-exploratory activity, the degree of severity of emotional reactions according to the "open field" test), M ± m in animals of different groups are presented in Table 2.
From the presented data it follows that after the animals were introduced to pain stress in both control groups ("no impact" and "placebo"), the main indicators of fear and anxiety significantly increased, as evidenced by the LPFM figures and especially the LPEC (by 44% and 43%, respectively). At the same time, it should be noted that in the "placebo" group, LPFM did not increase so sharply (by 0.2 s), which confirms the presence of a "placebo effect" in animals. In the experimental group of rats, as a result of the PPBM course, these indicators decreased significantly, approaching intact values. The data is illustrated with graphs (Fig. 1 a, b).
Indicators of horizontal activity of animals, which are expressed by the number of crosse
squares (NCS), the number of vertical racks and boluses, are also considered markers of stressful motor and orienting-exploratory behavior (Fig. 2-3a).
It is noteworthy that in the animals of the experimental group these indicators even exceeded the intact values, which confirms a decrease in their level of anxiety, fear and uncertainty. The intensity of grooming in the experimental group was also close to intact values, compared to an increase in the control (Fig. 3b).
A study of the dynamics of the microcirculation index showed that with damage to the sciatic nerve and concomitant pain syndrome, tissue perfusion decreased by 52% compared with intact animals, which is natural for the pathogenesis of pain injury. In the group with irradiation of acupuncture points of "general" and "local" action, a decrease in tissue perfusion was observed by 57% compared with the intact group of animals. The role of shunting blood flow (bypass index) increased only in the experimental group (by 15% of the intact values), and in the placebo group it remained at at the intact level. The ranges of fluctuations of active regulatory factors (endothelial, neurogenic, and myogenic components) were reduced in the placebo group by 23%, 17%, and 12%, in contrast to the growth in the experimental group (10%, 33%, and 15%) relative to intact animals, respectively. The level of microhemodynamic parameters in the Placebo group (control 3) did not differ statistically significantly from those of animals in the control 2 group without exposure (Table 3).
For passive regulation factors (respiratory and cardiac components), a different reaction was revealed, namely, in Control group 2, the respiratory component increased by 77% and the cardiac component decreased by 6%, while in the experimental group, the respiratory component decreased by 7% and the cardiac component by 24% compared to intact values, respectively (Table 3).
The data from the correlation analysis revealed a high degree of correspondence in the development of cognitive and metabolic disorders. In the Placebo group, a positive correlation was found between the concentration of
Table 1
Dynamics of oxidative metabolism indicators under conditions of pain stress
Biochemical indicators Control 1 (intact) Control 2 (pain stress without impact) Control 3 ("Placebo") Experience (pain stress with impact)
MDA in plasma, micromol/l 1.070 ± 0.009 0.864 ± 0.031 0.873 ± 0.029 0.790 ± 0.032**
MDA in erythrocytes, micromol/l 5.952 ± 0.071 8.627 ± 0.285* 8.594 ± 0.330* 7.995 ± 0.383**
DC in plasma, RVU 0.803 ± 0.013 0.893 ± 0.011* 0.908 ± 0.006* 0.888 ± 0.016***
TC in plasma, RVU 0.265 ± 0.004 0.325 ± 0.008* 0.312 ± 0.006* 0.288 ± 0.006***
SB in plasma, RVU 0.176 ± 0.007 0.212 ± 0.007* 0.206 ± 0.005* 0.189 ± 0.004***
SOD, RVU/min*mg of protein 1037.762 ± 21.351 962.834 ± 47.038* 971.656 ± 53.218* 1102.65 ± 79.78**
Catalase, RVU/min*mg of protein 39.512 ± 0.903 34.891 ± 1.603* 35.858 ± 1.970* 36.287 ± 1.491*
Notes: * - differences are statistically significant compared to "control 1" (intact rats) (p < 0.05); ** - differences are statistically significant compared to "control 2" (p < 0.05).
Table 2
Behavioral indicators in animals of different groups in the OP test
Groups LPFM, s LPEC, s NCS, pcs Racks, pcs Grooming, s Boluses, pcs
Control 1 (intact) 1.0 ± 0.09 65.0 ± 9.2 43.0 ± 3.4 4.8 ± 0.35 4.0 ± 0.55 1.0 ± 0.1
Control 2 (pain stress without impact) 2.5 ± 0.23* 300.0 ± 27.27* 22.0 ± 2.00* 0.5 ± 0.05* 22.0 ± 2.0* 0.1 ± 0.1*
Control 3 ("Placebo") 1.2 ± 0.11*** 280.0 ± 25.45*/** 35.0 ± 3.18*/** 2.0 ± 0.18*/** 15.0 ± 1.3*/** 0.6 ± 0.2*/**
Experience (pain stress with impact) 0.9 ± 0.08** 79.0 ± 7.18*/** 51.0 ± 4.64*/** 5.5 ± 0.50** 3.0 ± 0.2*/** 1.3 ± 0.4**
Notes: latent period of the first movement (LPFM), latent period of exit to the center (LPEC), number of crossed squares (NCS); * - reliability relative to the group of intact animals (p < 0.05); ** - reliability relative to the group of injured animals without exposure to PPBM (p < 0.05)
Table 3
Dynamics of microcirculation indicators in conditions of pain stress
Microcirculation parameters Control 1 (intact) Control 2 (pain stress without impact) Control 3 ("Placebo") Experience (pain stress with impact)
Microcirculation index, perfusion units 9.5 ± 0.73 4.3 ± 0.22* 4.5 ± 0.35* 4.2 ± 0.32*
Endothelial component, RVU 11.1 ± 0.85 8 ± 0.5* 8.5 ± 0.65* 12.2 ± 0.94**
Neurogenic component, RVU 8.7 ± 0.67 6.65 ± 0.46* 7 ± 0.54* 11.3 ± 0.87*/**
Myogenic component, RVU 8.4 ± 0.65 5.2 ± 0.37* 5.5 ± 0.42* 9.9 ± 0.75*/**
Respiratory component, RVU 7.2 ± 0.55 12.35 ± 0.95* 13 ± 0.99* 6.8 ± 0.52**
End of table 3
Microcirculation parameters Control 1 (intact) Control 2 (pain stress without impact) Control 3 ("Placebo") Experience (pain stress with impact)
Cardiac component, RVU 3.8 ±0.29 3.4 ± 0.13* 3.6 ±0.28 3 ± 0.23*
Bypass index, perfusion units 1 ±0.08 0.95 ±0.05* 1 ± 0.07 1.2 ±0.09*'**
Notes: * - differences are statistically significant compared to "control 1" (intact rats) (p < 0.05); - differences are statistically significant compared to "control 2" (p < 0.05).
3 2,5 2 1,5 1 0,5 0
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ll
350 300 250 200 150 100 50
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control 1 control 2 (intact rats) (pain stress without exposure)
control 3 ("placebo")
experience
control 1 control 2 control 3 experience (intact rats) (pain stress ("placebo") without exposure)
a)
b)
Fig. 1. Comparative data of latent period of first movement (LPFM) (a) and latent period of exit to the center (LPEC) (b) in OF tests in animals with pain stress.
Notes: * - reliability relative to animals in the intact group (p < 0.05); ** - reliability relative to the group of injured animals without exposure to PPBM (p < 0.05)
60
vi o
50 I ■ i 6
i40 H
130 ■
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control 1 control 2 control 3 experience control 1 control 2 control 3 experience
(intact rats) (pain stress ("placebo") (intact rats) (pain stress ("placebo")
without without
exposure) exposure)
a) b)
Fig. 2. Number of crossed squares (NCS) (a) and vertical racks (b) in OF tests in animals with pain stress. Notes: * - reliability relative to the group of intact animals (p < 0.05); ** - reliability relative to the group of injured animals without exposure to PPBM (p < 0.05)
1,6 1,4 1,2
| 0,8 ¡0,6 ¡S 0,4 0,2 0
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control 1 control 2 control 3 (intact rats) (pain stress ("placebo") without exposure)
experience
25
20
E 15
10
control 1 control 2 (intact rats) (pain stress without exposure)
control 3 ("placebo")
experience
a) b)
Fig. 3. Boluses (a) and grooming time (b) in OF tests in animals with pain stress.
Notes: * - reliability relative to the group of intact animals (p < 0.05); ** - reliability relative to the group of injured animals without exposure to PPBM (p < 0.05)
MDA in erythrocytes and LPEC (r = 0.571, p = 0.029), MDA in erythrocytes and grooming (r = 0.869, p = 0.013), SB in plasma and grooming (r = 0.902, p = 0.002), SB in plasma and LPEC (r = 0.764, p = 0.034), SOD activity and racks (r = 0.914, p = 0.008) and negative correlation between DC in plasma and racks (r = = -0.835, p = 0.017), SOD and LPFM activity (r = -0.698, p = 0.025), catalase and LPFM activity (r = -0.763, p = 0.022). Analysis of relationships revealed a positive correlation between MDA in erythrocytes and grooming (r = 0.772, p = =0.019), SB in plasma and LPFM (r = 0.805, p = 0.007) and a negative correlation between TC in plasma and racks (r = -0.912, p = 0.003), plasma SB and racks (r = -0.685, p = 0.026), catalase activity and LPEC (r = -0.749, p = 0.011) in the group of animals with pain stress without exposure.
In the experimental group of animals, there was a positive correlation between plasma DC and LPEC (r = 0.981, p = 0.001), plasma DC and grooming (r = 0.949, p = 0.001), SB in plasma and grooming (r = 0.977, p = 0.003), in addition, there is a negative correlation between plasma DC and racks (r = -0.975, p = 0.004), SOD activity and grooming (r = -0.911, p =
= 0.007), SOD activity and LPFM (r = -0.873, p = 0.012), catalase activity and LPEC (r = = -0.896, p< 0.001).
In the Placebo group, a positive correlation was revealed between the microcirculation index and the racks (r = 0.602, p = 0.036). In the group of animals with pain stress without exposure, the analysis of relationships revealed a positive correlation between the respiratory factor regulating microcirculatory blood flow and LPFM (r = 0.612, p = 0.028), cardiac factor and racks (r = 0.634, p = 0.030), and a negative correlation between the microcirculation index and LPFM (r = -0.583, p = 0.040).
In the experimental group of animals, there was a positive correlation between the respiratory factor regulating microcirculatory blood flow and grooming (r = 0.592, p = 0.025), in addition, a negative correlation between the endothelial factor regulating microcirculatory blood flow and LPEC (r = -0.588, p = 0.019).
Discussion
The study showed that a decrease in the concentration of lipid peroxidation products, an increase in SOD activity and, as a consequence, a decrease in the level of superoxide anions after
irradiation during pain stress had a positive effect on the cognitive function of rats. Our findings coincide with the opinions of other researchers. Thus, in the article (Zhang et al., 2023), based on an analysis of 150 literature sources, there is evidence of the positive effect of acupuncture on cognitive impairment in various pathological conditions due to an increase in the activity of SOD and catalase in the hippocampus of rats, whose neurons are damaged under pain stress. It has been noted that as a result of stimulation of acupuncture points, superoxide free radicals are converted into hydrogen peroxide. In doing so, catalase can prevent hydrogen peroxide from entering cells to form hy-droxyl free radicals, thereby reducing oxidative stress. It has also been demonstrated that AP stimulation activates a complex signaling pathway to reduce the production of ROS, MDA and other oxides. Coupled with an increase in the level of SOD and other antioxidants in hip-pocampal neurons, acupuncture can restore the oxidative-antioxidant balance of the brain with impaired cognitive functions (Thomas et al., 2009).
The results obtained confirmed the positive anti-stress effect of the PPBM on the body of animals in the experiment. Impact on the combination of points of "general" and "local" action against the background of pain stress led to a significant improvement in the orientation-exploratory behavior of experimental animals, a decrease in the level of fear, anxiety and optimization of the behavioral indicators of rats compared to the control, which confirms the adaptive mechanism of the sanogenetic effect of acupuncture (Yang et al., 2021).
The established correlation coefficients between microcirculation indicators and behavioral activity data during pain stress turned out to be less than the correlation coefficients between indicators of oxidative stress and cognitive functions, which confirms the determining role of impaired oxidative metabolism in the formation of cognitive impairment. The results obtained about the high degree of dependence of the motor and orientation-exploratory activity of animals on indicators of oxidative metabolism under the influ-ence of PPBM during
pain stress coincide with the results of other authors who conducted a study of the mechanism of action of acupuncture in cognitive and emotional disorders (Zhang et al., 2023; Thomas et al., 2009).
An increase in endothelial oscillations may be associated with increased release of endogenous nitric oxide. Myogenic fluctuations reflect the influence of central trophotropic mechanisms, including parasympathetic centers; their appearance in the spectrum of blood flow fluctuations indicates a decrease in the ergotropic central regulatory component and a shift in central regulation in the trophotropic direction. An increase in the amplitude of the respiratory wave indicates a decrease in microcirculatory pressure and / or a deterioration in venous outflow. Deterioration of outflow blood from the microcirculatory bed leads to an increase in the number of red blood cells, which is accompanied by an increase in the amplitude of the respiratory wave. A decrease in the amplitude of the pulse wave at elevated or normal values of average perfusion indicates a decrease in the flow of arterial blood into the microcirculatory bed (Bajrovic et al., 2000).
Conclusions
Our placebo-controlled experimental study confirmed the negative effect of the lipid peroxidation system on OS and behavioral activity of rats in the acute phase of pain stress. Analysis of the changed indicators revealed a clear correlation between the disturbance of oxidative metabolism processes and the degree of psycho-emotional disorders that arose. Subsequent correction of the resulting disorders by the course effect of PPBM on the AP confirmed the adaptive and stress-protective capabilities of laser puncture as one of the modern non-invasive, easily dosed, non-toxic effects on the body. This is confirmed by the statistically significant placebo-controlled positive dynamics of the studied indicators in the experimental group compared to the control, where after the end of the experiment significant violations remained compared to the initial values. The registered results indicate the stress-limiting effect of PPBM under experimental pain stress and con-
firm its analgesic effect in in vivo conditions. The demonstrated chains of interaction between various indicators of lipid peroxidation and parameters of behavioral activity of animals in the
process of adaptation of the body to acute pain stress allow us to recommend laser acupuncture technology for wider use in the complex rehabilitation of patients with pain syndrome.
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