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Khabibullayev B. B., Batyrbekov A. A.
Republican Scientific Center of Immunology of the Ministry of Health of the Republic of Uzbekistan, Tashkent E-mail: shga2065@yandex.ru
THE INFLUENCE OF HARMALA AND KARELINIA CASPIA ON THE IMMUNE STATUS FOR RAY DISEASE IN EXPERIMENT
Abstract: It was established, that Harmala and Karelinia caspia possess ability increase immune response to sheep erythrocytes and titer antierythrocytes antibodies in peripheral blood, stimulated proliferation cells in central (thymus, bone morrow) and peripheral (lymph nodes) organs of immunity of mice radiation sickness.
Keywords: Harmala, Karelinia caspia, radial illness, immune response, organs of immunity, erythrocytes, leukocytes.
Currently, plants attract the close attention ofresearch-ers as a source of obtaining biologically active drugs with different vectors ofeffects on the body. A rich source ofob-taining herbal medicines is the Central Asian region [9]. Herbal preparations are used for many somatic [2, 4], oncological diseases [5, 7], liver diseases [6], with radiation effects [3, 8]. Many plants have hypoglycemic activity [1].
The aim of the work is to study the influence of the hypoglycemic plant-based means (Peganum harmala) and Karelinia caspia (Karelinia caspia) on immunogenesis in radiation sickness.
Material and methods
In the experiments, white mongrel mice of2-3 months of age weighing 20-22 g were used. For modeling radiation sickness (RS), mice were irradiated in total at a dose of 5 Gy. After 5 days, they were intraperitoneally immunized with sheep red blood cells (SRBC) at a dose of 2 x 108
and after 5 days the number of antibody-producing cells (ABPC) of spleen was determined [10]. There also were determined the titer of antibodies to SRBC in the blood, the number of cells in the thymus, in the bone marrow, in the mesenteric lymph nodes, the number of erythrocytes and leukocytes in the peripheral blood of mice.
Results and discussion
The results of studies on the effect of harmala and Karelinia caspia on immunological reactivity in mice with RS are given in Table 1. As can be seen from this table, in the spleen of the mice of the control group an average of 7756.3 ± 366.4 antibody-producing cells are formed, and in animals with RS in 10, 3 times less. Consequently, after the radiation exposure, a deep secondary immunodeficiency is formed. It is established that all the studied herbal remedies have the ability to correct to a certain extent violations in the immune status in irradiated mice.
Table 1. - Effect of Harmala and Karelinia caspia on the immune response to sheep red blood cells in mice with radiation sickness (RS) (M ± m)
Dose. mg/ kg Number of NCS x 106 The number of ABPC at
Group IR whole spleen IR 106 cells of the spleen IR
1 2 3 4 5 6 7 8
1. control (n = 8) - 163.6 ± 3.9 - 7756.3 ± 366.4 - 47.6 ± 2.6 -
2. RS (n = 8) - 39.9 ± 0.9a -4.10 756.3 ± 32.0a -10.3 19.0 ± 1.0a - 2.51
Section 7. Medicine
1 2 3 4 5 6 7 8
3. RS + Harmala (n = 8) 50.0 71.8 ± 1.7ab +1.80 2681.3 ± 113.8ab +3.55 37.5 ± 1.9ab + 1.97
4. RS + KC (n = 8) 100.0 59.9 ± 1.4abc +1.50 2206.3 ± 89.4abc +2.92 37.0 ± 1.8ab + 1.95
5. RS + KC + veh. (filler) (n = 8) 100.0 63.9 ± 1.5abc +1.60 2431.3 ± 102.6ab + 3.21 38.2 ± 2.0ab + 2.01
Note: here and in the following tables, KC is Karelinia caspia, KC + veh. (filler). - Karelinia caspia with vehicle (filler), NCS - nucleated cells of the spleen; ABPC - antibody-producing cells; IR - the index of the ratio: (-) - in relation to 1 gr., (+) - in relation to 2 gr., A - authentically to 1 gr., B - authentically to 2g., C - authentically to 3g.
In mice receiving Harmala diet, 2.681.3 ± 113.8 ABPC are formed in the spleen, which is significantly 3.55 times higher than in the immunodeficiency group. In animals with RS who received Karelinia caspia, the immune response to sheep blood cells is 2.92 times higher than in untreated mice. Immunostimulating activity of Harmala is significantly higher than in Karelinia caspia. Kareliania Caspian with a filling of 3.21 times increases the depressed number ofABPC in mice with RS. Thus, the studied herbal remedies have the ability to some extent to increase the immune response to SRBC in irradiated animals. When calculating AOC for 1 million spleen cells, the following results were obtained. In the control group, this indicator is 47.6 ± 2.6 ABPC, and in mice with RS - 2.51 times less.
Under the influence of Harmala, this indicator is 1.97 times higher compared to the untreated group, the Karelinia caspia - by 1.95 times, the Karelinia caspia with vehicle (filler) - by 2.01 times. There is no significant difference between the indicators. Thus, when calculating the ABPC for both the whole spleen and 1 million sple-
nocytes, the ability of the studied herbal remedies to significantly enhance immunological reactivity in mice after radiation exposure was established. As can be seen from Table 1, the total number of NCS in the control is 163.6 ± ± 3.9 '106, and after the radiation exposure, this index decreases by 4.10 times. When the Harmala is administered, the total number of splenocytes in mice with RS significantly increases in comparison with the untreated group by 1.80 times. With the introduction of Karelinia caspia, the number of NCS in irradiated mice increases 1.50 times. In the group receiving Karelinia caspia with vehicle (filler), the number of splenocytes increases by 1.60 times. The stimulating activity of harmal is significantly higher than in Karelinia caspia without and with a reservoir. Thus, the studied herbal remedies have the ability to increase the immunological reactivity of the organism and the total number of NCS in mice with RS.
Further we investigated the effect of herbal remedies for antibody titer to SRBC in the peripheral blood of mice with RS (Table 2).
Table 2.- Influence of Harmala and Karelinia caspia on the titer of antibodies to erythrocytes of the sheep in the blood of mice with radiation sickness (RS) (M ± m)
Group Dose. mg / kg Titer of antibodies to SRBC (log2) IR
1. control (n = 8) - 5.4 ± -
2. RS (n = 8) - 2.0 ± -2.70
3. RS + Harmala (n = 8) 50.0 2.8 ± + 1.40
4. RS + KC (n = 8) 100.0 2.5 ± + 1.25
5. RS + KC + veh (filler) (n = 8) 100.0 2.6 ± + 1.30
On the 4 th day after immunization, the antibody titer to SRBC in the control group is 5.4 ± 0.2, and in irradiated mice is 2.70 times less (p < 0.05). When administered to mice with RS Harmala, the titer of antibodies
to SRBC in the blood is significantly increased by 1.40 times in comparison with the previous group. Kareliania caspia without vehicle (filler) increases the antibody titer to SRBC by 1.25 times, and with the vehicle (filler) - by
1.30 times. However, in both cases, the difference with the irradiated and untreated group proved to be unreliable. Consequently, only under the influence of Harmala there is a significant increase in the titer of antibodies to SRBC in mice with RS.
Next, we studied the effect of herbal remedies in the state central and peripheral organs of immunity system in mice with RS. The results of these studies are presented in (Table 3).
Table 3.- The Effect of Harmala and Karelinia caspia on the number of cells in immunity organs in mice with radiation sickness (RS) (M ± m)
Group Dose, mg/ kg Thymus' cells x 106 Cell IR Bone marrow cells IR Lymph nodes cells f IR
1. control (n = 8) 39.4± - 12.7± - 25.3± -
2. RS (n = 8) - 12.7± -3.10 4.5± -2.82 9.4± -2.72
3. RS + Harmala (n = 8) 50.0 17.1± +1.35 6.2± + 1.38 12.1± +1.29
4. RS + KC (n = 8) 100.0 16.2± +1.28 5.7± + 1.27 11.0± +1.17
5. RS + KC + veh. (filler)(n = 8) 100.0 16.7± +1.32 5.9± + 1.31 11.5± +1.22
As can be seen, the total number of cells in the thymus of the control group mice is 39.4 ± 1.7 x 106, and in animals with RS it is 3.10 times smaller. When irradiated mice of harmala, Karelinia caspia with vehicle (filler) are injected, the number of cells in the thymus significantly increases 1.35 times, 1.28 times and 1.32 times, respectively. The stimulating activity of the studied samples does not differ significantly from each other. Consequently, all studied plant products are able to increase the number of cells in the thymus in mice with RS. The total number of cells in the bone marrow in irradiated mice is reduced by 2.82 times. Under the influence of Harmala, the number of cells in the bone marrow increases significantly by 1.38 times. In irradiated mice that received Karelinia caspia, the number of bone marrow cells significantly increased by 1.27 times in comparison with the untreated group.
The stimulating activity of the Karelinia caspia is significantly lower than in the harmaly. In the group of mice with RS, who received Karelinia caspia with vehicle (filler), the number of cells in the bone marrow increases by 1.31 times (p < 0.05), and this level does not differ significantly from the data obtained in animals receiving Harmala. Thus, one can draw a conclusion about the ability of the studied agents to stimulate the proliferation of cells in the central (thymus, bone marrow) immunity organs in irradiated animals.
Next, the effect of plant remedies on the total number of cells in the peripheral organs of immunity (mesenteric lymph nodes) was studied in mice with RS (Table 3). In the control group, the number of cells in the lymph nodes is 25.3 ± 0.8'106, and in irradiated mice is 2.72 times less. All studied herbal remedies significantly increase the number of cells in lymph nodes in mice with RS: harmala - 1.29 times, Karelinia caspia - 1.17 times Karelinia caspia with filler - 1.22 times. By stimulating activity, the studied samples do not differ significantly from each other. Thus, all the studied herbal remedies have the ability to increase the number of cells in the central and peripheral organs of immunity in irradiated animals.
The results of studies on the evaluation of the effect of herbal remedies on hematologic indices are given in Table 4. In the control group, the number of red blood cells is 4.7 ± 0.1 x 109 / ml, and in irradiated mice is 2.76 times less. In irradiated mice receiving Harmala, the number of erythrocytes in the blood increases significantly by 1.24 times. Under the influence of Karelinia caspia, the number of erythrocytes increases by 1.12 times (p < 0.05). The stimulating activity of harmal is significantly higher than that of Karelinia caspia. Addition of vehicle (filler) to Karelinia caspia increases its stimulating properties: the number of erythrocytes increases by 1.18 times. Consequently, the studied herbal remedies can in-
Section l. Medicine
crease the number of erythrocytes in peripheral blood the ability to reliably increase the number of leukocytes
in animals with RS. in the blood of irradiated mice.
The number ofblood leukocytes in irradiated mice is reduced by 2.88 times. The studied herbal remedies have
Table 4.- The Influence of Harmala and Karelinia caspia on the number of red blood cells and leukocytes in peripheral blood in mice with radiation sickness (RS) (M ± m)
Group Dose. mg|kg Erythrocytes x 109|ml IR Leukocytes x 106|ml IR
1. control (n = 8) - 4.7 ± - б.9 ± -
2. RS (n = 8) - 1.7 ± -2.7б 2.4 ± -2.SS
3. RS + Harmala (n = 8) 50.0 2.1 ± +1.24 3.0 ± +1.25
4. RS + KC (n = 8) 100.0 1.9 ± +1.12 2.S ± +1.17
5. RS + KC + veh. (filler) (n = 8) 100.0 2.0 ± +1.1S 2.9 ± +1.21
Under the influence of Harmala, the number of leu- The stimulating activity of Harmala is significantly higher kocytes increases by 1.25 times (p < 0.05) and Karelinia than that of Karelinia caspia. caspia with vehicle (filler) - by 1.21 times (p < 0.05).
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