[Effect of inhibitors of cell metabolism on postradiation repair and degradation of DNA in rat thymocytes].
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Biomedical subjects
Publications and source records attributed to N I Riabchenko.
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Exposure to radiation, as well as holding under conditions of limited mobility during 24 h, induced decrease in thymus cell number, increase in number of DNA breaks. The content the products of lipid peroxidation reactive with thiobarbituric acid in blood serum of mice decreased as well. The stress effect is comparable with radiation doses in the range of 50-60 cGy.
The animals were irradiated within adaptive dose of 0.1 Gy and 5 hours later with a challenge dose of 2 Gy. The adaptive dose reduced the effects induced by the challenge dose of 2 Gy: the increased content of the products of lipid peroxidation reactive to thiobarbituric acid in blood serum, and the increased number of breaks in thymus DNA of irradiated mice.
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Apoptosis in peripheral blood lymphocytes of healthy donors and cancer patients after gamma-irradiation with different doses was studied by the flow cytometry method. Wide intra- and interindividual variabilities of the lymphocyte radiosensitivity were observed. The radiosensitivity did not depend on the subpopulation composition of the lymphocyte pool. The persons with very low and high lymphocyte radiosensitivities were found significantly more often among the cancer patients than among the healthy donors. One can suggest that this method is useful as a biomarker of future cancer risk and prognosis of radiotherapy efficiency.
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Accumulation and excretion of beta-carotene in rat liver tissue were studied when the carotinoid was intraperitoneally administered as olive oil as suspension and a water liposomal suspension. Single injection of the liposomal form of beta-carotene caused an increase in its content in liver tissue. If beta-carotene was administered at a dose of 10 mg/kg its maximal content 44 mg/g of the tissue was observed within one day, and at a dose of 50 mg/kg, its maximal content constituted 178 mg/g within two days. Administration of beta-carotene as an oil suspension at a dose of 50 mg/kg led to a considerably slight accumulation in rat liver tissue with the maximal value of 12 mg/kg of the tissue within two days. Possible use of beta-carotene various forms, as well as experimental models for study their biological activity are discussed.
The influence of a tumor necrosis factor, administered 16 h before irradiation of rats, on the radiation response of thymus and bone marrow cells has been investigated. Three and 6 h after irradiation the following indices were analyzed: the number of apoptotic cells in the thymus; the accumulation of polydeoxyribonucleotides and the appearance of single-strand breaks in DNA of bone marrow and thymus cells; and the electrophoretic properties of thymocyte DNA. The injection of a tumor necrosis factor reduced the number of polydeoxyribonucleotides, inhibited internucleosome DNA fragmentation, and did not influence the formation of single-strand breaks in DNA.
Human peripheral blood lymphocytes were exposed to 60Co gamma-rays (a dose of 3 Gy) and cultivated during seven days in the presence of PHA and BrdU. It was shown that the metaphases of the first and second mitosises occurred during cultivation of the irradiated and unirradiated lymphocytes, being evidence about of irregularity of the coming into division of various fractions of lymphocytes. The time of cultivation did not influence a rate of aberrations in metaphases of the first and second mitosises of the irradiated lymphocytes. During the first and the subsequent mitosises the number of exchange chromosome aberrations decreased and reached a control level in metaphases of the fourth and fifth mitosises. The number of paired fragments at second and third mitosises increased a little and started to decrease only in metaphases of the fourth and fifth mitosises. The decrease in chromosome aberrations with prolongation of the cultivation of lymphocytes after irradiating is a consequence of elimination of cells with chromosome damages during sequential mitotic divisions.
The radioprotective and antistressful activities of L-arginine and the "Pronumol" preparation, in which L-arginine is contained in the complex of proteins with nucleic acids, were studied. In mice repeated peroral intake of L-arginine and "Pronumol" partially prevented radiation-induced and stress-induced lipid peroxidation and DNA degradation in thymus, increased hemopoietic stem cell survival, and prevented an increase in chromosome aberration frequency in bone marrow cells of irradiated mice. When repeatedly administered per os before irradiation, "Pronumol" increased survival of intestinal stem cells in irradiated mice and prevented thymus cell devastation induced by radiation and stress.
The V-79 Chinese hamster cells were irradiated by gamma-rays in dose of 0.5 Gy at powers of doses 0.48 Gy/min (an acute irradiation) and 0.0485 MGy/min (a prolonged irradiation). The acute and prolonged irradiation in a dose of 0.5 Gy enlarges frequency of the appearance of micronucleus (MN). Subsequent cultivation of the irradiated cells during 20 generations enlarges frequency of MN, and for prolonged an irradiation the boosted frequency of MN, is saved during 40-60 generations. After an acute irradiation the number of MN starts to reduce after 20 doublings.
Solutions of the ferrous sulfate, of the albumin and of the suspension of liposomes were irradiated by mixed gamma-neutron radiation (fission spectrum neutrons, contribution of gamma-component to the absorbed dose up to 20%) at the pulse reactor BARS-6 with single-pulse (duration 100 micros) or continuous radiation (duration 60 min). It was shown, that after the pulse irradiation the concentration of the malonic dialdehyde in liposomes was in 3-4 times higher than after the continuous radiation at equal absorbed doses (p < 0.05). On the contrary, the irradiation of the liposomes suspension as well as of the solutions of the ferrous sulfate and of the albumin in a mode of single-pulse or of continuous mode did not reveal the statistically significant differences in the production of Fe3+ ions and of peroxides of the albumin for two mode of the radiation action.
The irradiation with mixed gamma-neutron radiation was carried out at the pulse nuclear reactor on fast neutrons BARS-6 in a regimen of one pulse (100 micros) and in a regimen of continuous irradiation during 60 minutes. Was shown, that the irradiation of mice with pulse radiation was 1.3-1.8 times more effective in the induction of the chromosome aberrations in bone marrow cells in comparison with the continuous regimen of irradiation. At the same time, other biological tests (yield of chromosome aberrations in human lymphocytes, decreasing the number of cells in thymus) demonstrated that pulsed and continuous regimens have almost equal biological effectiveness.
Regularities of chromatin degradation in thymocytes and LS/BL tumor cells have been investigated. It has been shown that the rate of DNA degradation by Ca/Mg-dependent endonuclease in LS/BL tumor cells is 25 times lower than that in thymocytes, and radiation does not induce chromatin degradation. The alkylating agent TS 160 causes chromatin degradation in both LS/BL cells and thymocytes. In contrast to radiation TS 160 inhibits the endogenous chromatin degradation by Ca/Mg-dependent endonuclease in thymocytes.
The administration of dextran sulfate causes an increase in the thymidine content of rat blood comparable with the postirradiation thymidinemia. In contrast to radiation dextran sulfate does not increase polydeoxynucleotide content of thymus. Isoptin, a calcium antagonist, decreases thymidinemia induced by dextran sulfate to a greater extent than that induced by ionizing radiation. Thymidinemia induced by dextran sulfate is supposed to be due to its effect on cell membranes. In radiation-induced thymidineamia this mechanism is of lesser significance.
Total DNA and DNA of chromatin degradation products obtained from rat thymocytes 6 h after irradiation with a dose of 10 Gy were separated electrophoretically. Relative shares of mononucleosomes and their oligomers were determined. Experimental distributions of DNA fragments differ from those calculated on the basis of the assumption of a random breakage of bonds between the nucleosomes.
It was shown that in conditions optimal for Ca/Mg endonuclease, chromatin endonucleolysis in the nuclei and thymocytes occurs due to internucleosome fragmentation of DNA. Irradiation activates chromatin degradation in thymocytes washed by a buffer containing 0.25 M sucrose, 10 mM tris-HCl, pH 7.2, 3 mM MgCl2, and does not influence this process in thymocytes washed by 10 mM tris-HCl, pH 7.2, 3 mM MgCl2.