RU2002265C1 - Method of blood treatment - Google Patents

Abstract

The invention relates to medicine, more specifically to hematology, and can be used to improve the structural and functional state of membranes, in particular, red blood cells. The aim of the invention is to improve the functional state of the erythrocyte membrane. SUMMARY OF THE INVENTION Blood is treated with laser radiation in the yellow spectral region with a wavelength of 578 nm in a pulsed-periodic mode with a light -8 pulse of 15x10 and a pulse repetition rate of 8 kHz at a dose of 1 J per t cm . The effectiveness of improving the structural and functional state of cell membranes was evaluated by the interaction of the system / lipid peroxidation / lipid peroxidation-antioxidant activity / AOA /

Description

The invention relates to medicine, more specifically to hematology. Normal membrane function, & erythrocytes, in particular, plays an important role in the vital activity and homeostasis of the body. Violations of the structural and functional state of erythrocyte cell membranes are associated with a pathological process in the body.

There are analogues of a blood processing method for the correction of plasma membranes, in particular, red blood cells with tocopherols, vitamin D, Aevit preparations.

However, these chemicals can have various indirect effects, as well as being metabolized and excreted in time.

A known method of treating blood by irradiating with laser radiation in the red region of the spectrum of native donor blood or by intravenous irradiation of blood, i.e. blood is irradiated in two ways - extracorporeal and intravascular. The similarity of the results of these methods due to the fundamental physical community of instrumental approaches and, most importantly, the similarity of the induced biological effects is proved. It should be borne in mind that helium-neon laser radiation acts on the body mainly through the eyebrow. It was established that upon irradiation of blood, a distinct protective effect of helium-neon laser radiation is observed in preserving the structure of erythrocyte membranes or in correcting its changes caused by the pathological process. However, physiological correction requires optimization of radiation doses, since if critical doses are exceeded, a transition from stimulation to irreversible damage to the membrane structure is possible. Helium-neonose laser radiation affects lipid peroxidation processes.

Despite the protective nature of the protection of helium-neon laser erythrocyte membranes and the improvement of donor blood properties, there are the following disadvantages: the exposure to laser radiation in the red region of the spectrum is continuous for a long time; irradiation does not completely restore the phospholipid component in the erythrocyte membranes; the structural and functional state of the membranes is not restored: a) lipid peroxidation is sharply increased, b) antioxidant activity is reduced, which creates a low protection of cell membranes from destruction; the lipid composition of the membranes changes, causing the loosening of the membranes, which leads to increased sensitivity of phospholipids to peroxidation.

The purpose of the invention is to improve the functional state of erythrocyte membranes.

The goal is achieved by irradiating blood or erythrocyte mass with laser radiation in the yellow region of the 578 nm long wavelength spectrum with a light pulse duration of 1.5 10 s, a pulse repetition rate of 8 kHz, an average output power of 15 mW and an average power density of 2 W / cm2 at a dose of 1 J per 1 cm. Irradiation is carried out from a Yakhrom laser using copper vapor.

The functional state of the membranes was assessed by the ability of the cell to peroxidate hemolysis after irradiation and after incubation of a blood sample under physiological conditions (Tris-HC buffer pH 7.4, with the addition of KCI and NaCI at T 37 ° C with free access of oxygen); accumulation of lipid peroxidation metabolite (LPO) - malondialdehyde (MDA) after irradiation and incubation of samples; the ability of red blood cells to degrade malondialdehyde (D MDA) and the ratio of the degradation rate of malondialdehyde to the total content of maldealdehyde, which indirectly indicates the activity of aldehyde dehydrogenase and the ability of a cell to get rid of MDA: total antioxidant activity (AOA) and the ratio of AO A / MDA, which indicates the protection of the cell from peroxidation.

Blood was taken from a group of pregnant women with late toxicosis of varying severity in order to track and improve the correction effect of impaired metabolic processes (lipid peroxidation, AOA, the rate of degradation of MDA) as a result of irradiation of red blood cells with various laser radiation spectra and to identify their differences. We studied the red spectrum in a continuous mode — a wavelength of 632.8 nm, in pulsed modes — the red spectrum — a wavelength of 628 nm, the yellow spectrum — 578 nm, and the green spectrum — 510 m. The number of samples was 56 crocs and six women after surgery with complications.

It was established that in erythrocytes of women with pathology, abnormalities were detected, which can be described as imbalance of the LOL-AOA system. This fact is evidenced by the following data: increased mechanical hemolysis, indicating an increase in the viscosity and stiffness of the membrane, as a result of an earlier increase in

them FLOOR; sharply reduced antioxidant activity and antioxidant protection, creating conditions for raising lipid peroxidation and destruction of cell membranes; MDA was increased after incubation (by 47%) and the intensity of MDA degradation was 42.5%, which is typical for membrane loosening and greater accessibility of its phospholipids to ROL, i.e. some disorganization of cell membranes is possible.

A prototype method was used to correct violations of the plasma membranes of red blood cells: a red spectrum in a continuous mode from helium-neon laser radiation. Irradiation of a 5% suspension of red blood cells led to slight changes in hemolysis and MDA content; incubation of the sample led to an even greater increase in both peroxide hemolysis and MDA content (by 57 and 52%, respectively), which indicates a rise in LPO activity after irradiation of red blood cells with this laser spectrum; the rate of degradation of MDA has not changed; the ratio of DMDA / MDA decreased, indicating a lower ability of the cell to get rid of MDA and the possibility of accumulation of this metabolite; antioxidant activity increased slightly; the ratio of AOA to MDA has not changed.

Thus, despite a slight increase in AOA, the remaining indicators show that the LPO intensity even increases somewhat after exposure to the red laser spectrum in a continuous mode, increasing the imbalance in cell membranes.

When analyzing the results of the action of -various spectra on erythrocyte membranes, the advantage of the yellow spectrum was revealed, which is offered to improve the functional state of the membranes and to correct violations of the structural and functional organization of cell membranes.

The proposed method.

Irradiation of the sample with a 5% suspension of red blood cells taken from pregnant women with late toxicosis, a yellow spectrum with a wavelength of 578 nm in a pulsed mode led to: a decrease in hemolysis before incubation to normal values; an increase in hemolysis after incubation to a lesser extent (by 40%} than upon irradiation with the red spectrum in a continuous mode: there is no increase in MDA both before and after incubation, which allows us to judge the good functioning of all structures of plasma membranes and better degradation of MDA a erythrocytes and better protection of phospholipids from lipid peroxidation to increase the intensity of degradation of MDA and AOA, respectively, with a content of 5 MDA. not the ratio of D / MDA and AOA / MDA. indicating the physiological effect of laser radiation in the yellow spectrum on metabolic processes.

0 The results of the two methods are presented in the table, which shows that the laser radiation in the yellow region of the spectrum acts on the pathologically altered erythrocyte membrane more physiologically

5 than the red spectrum in continuous mode. To determine the optimal parameters of laser radiation, doses of 1 J and 2 J per 1 cm3 of solution were studied. It was found that irradiation with a laser beam in

0 yellow spectral region at a dose of 2 J leads to a sharp increase in the content of MDA (from 1.73 to 2.35 nmol / 106 er) and the indicator of degradation of MDA (from 49.3 to 65.9%) with a simultaneous decrease in AOA and antioxidant protection

5 (respectively, from 0.97 to 0.83 and from 0.56 to 0.36). The facts obtained indicate the depletion of antioxidant protection, which, when repeated exposures can lead to an imbalance of the LPO-AOA system,

0 deterioration in the functional state of cell membranes. A dose of 1 J acts on the membrane softer, more physiologically: the concentration of MDA decreases (from 1.26 to 0.79 nmol / 106 er) before incubation; increases

5 the degree of MDA degradation (from 37.5 to 41.3%) and the ratio of MDA degradation to the concentration of MDA (from 29 to 52); significantly increases AOA (from 1.74 to 3.66 μmol Ig 106 er. for 1 min) and the ratio of AOA to MDA (from 1.6 to

0 4.21). Therefore, a dose of 1 J leads to a better correction of metabolic processes in the cell membrane and can be used in various diseases, including pathology

5 pregnant women.

Initial background of the state of the patient erythrocyte membranes: hemolysis spontaneous before incubation 4.1%. after incubation - 4.6% (increase of 12%); MDA concentration before incubation 1.13 n / mol 106 er; after - 2.91 n / mol 106 er. (an increase of 148%), which indicates a sharp deterioration in the functional state of the membranes; degradation rate of 39.7%; ratio indicator

5 degradation to the concentration of MDA 35.1%, which is higher than normal. This indicates a sharp deterioration in the structural and functional state of cell membranes; AOA 1.19 mk

mol 10 er. in 1 min and antioxidant protection 1.05,

IRRADIATION DOSE 1 J per 1 cm3 Example 1. (prototype method). A patient erythrocyte suspension was irradiated with a red laser spectrum with a wavelength of 632.8 nm in a continuous mode from a helium-neon unit. Irradiation dose 1 J, exposure time 30 min, State of the cell membranes: spontaneous hemolysis before incubation 5.5% (slightly increased compared to the initial background, after incubation 7.1. I.e. an increase of 29%; MDA concentration before incubation 1 , 00 nmol / 106 er., After 2.25, which indicates a huge increase (125% increase) and thereby a sharp deterioration in the functional state of erythrocyte membranes; degradation rate

43.1%, which indicates an almost double increase in comparison with the physiological course of pregnancy - 21.9 ± 2.24%; the ratio of DMDA / MDA is 43.1, while normal

20.2 ± 3.28%, which indicates a sharp deterioration in the structural and functional state of cell membranes; the antioxidant activity was sharply reduced - 1.21 μmol 106 er, and the antioxidant Ed protection (AOA / MDA 0.49) in 1 min. It can be seen from the above example that irradiation with the red spectrum in the continuous mode practically does not contribute to membrane restoration.

Example 2 (the proposed method). Patient-C erythrocyte suspension was irradiated. laser radiation in the yellow spectral region with a wavelength of 578 nm, a pulse duration of 1.5 s, a repetition rate of 8 kHz at a dose of 1 J, exposure time 100 s.

The state of erythrocyte membranes: hemolysis of irradiated blood before incubation decreased almost to normal compared to the initial background - 0.94, after incubation - t, 3, a 40% increase; MDA concentration before incubation 1.33 nmol / 10 er., after - 1.5, i.e. practically unchanged, which is a positive point in stabilizing the state of the membranes; the rate of degradation is higher and amounted to 54%, which indicates the physiological effect in relation to the metabolic process; the ratio of DMDA / MDA 36%; AOA increased -2.11 μmol 10 er.for 1 min, and the antioxidant had a protection of 1.4.

The example shows that the yellow spectrum at a dose of 1 J in the pulsed mode has a positive effect on lipid peroxidation and AOA of cell membranes.

IRRADIATION DOSE 2 J on 1 cm3. Example 3 (prototype method). A patient C. erythrocyte suspension was irradiated with a red laser spectrum continuously from a helium-neon unit, dose 2 J. Exposure time 60 min.

The state of erythrocyte membranes: hemolysis after irradiation before incubation 1.45%, after - 2.09%, i.e. the increase was 44.1%; MDA concentration of 2.73 nmol / 106 er., after incubation, 2.73, i.e. not changed;

the degradation rate was high 48.6%; the DMDA / MDA indicator is below the norm - 17.9: the AOA indicator is reduced - 1.7 μmo / 10 er. in 1 min; the rate of antioxidant protection was also not high - 0.62.

It can be seen from the example that increasing the dose leads to a slight improvement in the condition of the membranes.

Example 4 (the proposed method). Irradiation of a suspension of red blood cells of the patient

C. at a dose of 2 J with a yellow spectrum of laser radiation with a wavelength of 578 nm, a pulse duration of 1.5 s, a pulse repetition rate of 8 kHz, an exposure time of 200 s.

The state of cell membranes: hemolysis before incubation, 1.51%, after - 1.86%, i.e. an increase of 23%; the concentration of MDA before incubation 2.35, after - 1.20 nmol / 106 er; MDA degradation rates and D ratio

MDA / MDA were high (65% and 55%, respectively), indicating an increased ability of red blood cells to degrade exogenous MDA; AOA 2.3, and the ratio of AOA / MDA 1.9.

It can be seen from the example that increasing the dose did not lead to a sharp improvement in the state of the membranes, but only led to the stress of the POL-AOA system.

Thus, the proposed method

blood processing is the most physiological and best in the correction of plasma membranes compared to the red spectrum in continuous mode. Achieving the goal occurs when the exposure time is reduced by 15 times, due to the optimization of the exposure parameters, and

namely: the yellow spectrum in pulsed mode at a dose of 1 J per 1 cm3.

55

(56) Gamale N.F. Abstract of the All-Union Conference: The effect of low-energy laser radiation on blood. Kiev: 1989.S. 180-182.

The action of laser radiation in the red region of the spectrum in continuous mode and in the yellow region of the spectrum in pulsed mode on lipid peroxidation sites. characterizing the state of the plasma membranes of red blood cells

P 0 05 J, - with a breakdown without incubation (breakdowns without irradiation - with a breakdown in the prototype method

Claims (1)

The claims METHOD FOR PROCESSING BLOOD by exposure to low-intensity laser radiation, characterized in that, in order to increase efficiency and reduce exposure time to maintain and restore functional state neither erythrocyte membranes are exposed to blood by laser radiation in the yellow spectral region with a wavelength of 578 im in the intermittent periodic quasi-intermittent mode with a light pulse duration of t, 5 s and a pulse repetition rate of 8 kHz at a dose of 1 J per t cm3.