The effect of combined application of quercetin and indralin on postradiation repair of the hematopoietic system in acute radiation sickness

Hybrid F1 mice (CBA × C57Bl/6) were subjected to irradiation at a nonlethal dose of 6.7 Gy that brought on acute radiation sickness. In the experiments performed, we observed the beneficial effect of combined application of quercetin injected 30–60 min prior to irradiation with γ rays and the emergent radioprotector indralin injected after irradiation on development of postradiation repair in hematopoietic tissue. This effect was expressed as accelerated formation of endogenous spleen colonies and recovery of spleen weight and attenuation of leucopenia 12 and 16 days after acute irradiation. Treatment with only quercetin was not radioprotective.     

Antibody-forming capacity of mouse spleen cells after hypoxic hypoxia and the administration of erythropoietin

In CBA mice the absolute and relative (per 10(6) spleen cells) number of antibody-forming cells (AFC) in the spleen was cut by half on the 1st, 4th, and 7th days after acute hypoxia (12 hours, 6700 m), and on the 1st and 4th days after cessation of chronic hypoxia (16 days, 16 hours, 6700 m). The number of AFC in the spleen returned to the normal level on the 7th day after cessation of chronic hypoxia. Single or double erythropoietin injections caused approximately a 1.15--2-fold decrease in spleen AFC number in posthypoxic mice in comparison with control animals.     

Radioprotective properties of indralin combined with cystamine and mexamine

The study of indralin radioprotective properties at its joint application with cystamine and mexamine was carried out in the experiments on inbred mice and rats. The mice and rats were exposed to whole-body y-irradiation at a dose of 9.0 and 9.5 Gy, correspondingly. A combined parenteral administration ofindralin and cystamine at a dose of 25 mg/kg showed ponentiaton of indralin radioprotective properties up to a level of the ED50 effect versus the absence of or a weak radioprotective effect in the case of their separate application. In the experiments on rats, indralin (50 mg/kg) and mexamine (12 mg/kg) injected intraperitoneally almost completely eliminated the animal mortality from the intestinal syndrome of acute radiation sickness amounting in the control radiation group to 60% on the 7th day after exposure to radiation at a dose of 9.5 Gy. However, at the above conditions, radioprotectors at these doses had a low-level radioprotective action at the onset of the bone marrow syndrome of acute radiation sickness. Combined application of indralin and mexamine at the same doses and at the same conditions led to a radiation protection 50% as high as in the case when radioprotectors were applied separately at a double dose.     

The cytokinesis-block micronucleus assay as a biological dosimeter in spleen and peripheral blood lymphocytes of the mouse following acute whole-body irradiation

To evaluate the application of the cytokinesis-block (CB) micronucleus (MN) assay as a biological dosimeter following in vivo exposure to ionising radiation we determined the micronucleus frequency in spleen and peripheral blood lymphocytes of the mouse, serially, for 14 days following acute whole-body irradiation. The baseline MN frequency of spleen lymphocytes (7.86 +/- 0.68, mean +/- 1 SD) was significantly (p less than 0.001) elevated when compared to that for peripheral blood lymphocytes (4.10 +/- 0.53). Immediately after irradiation there was a substantial dose-related increase in MN, but the MN frequencies in spleen lymphocytes (120.2 +/- 9.4 for 1 Gy; 409.5 +/- 38.4 for 2 Gy) were significantly (p less than 0.009) elevated compared to those in peripheral blood lymphocytes (78.0 +/- 7.0 for 1 Gy; 200.2 +/- 10.9 for 2 Gy). During the 14 days after irradiation, the MN frequency in spleen lymphocytes declined gradually to approximately half of the value observed immediately after irradiation. By contrast the MN frequency in peripheral blood lymphocytes increased during the week after irradiation, but ultimately MN frequencies in blood and spleen became approximately the same by day 14. Study of isolated murine lymphocytes irradiated in vitro showed that the number of MN generated by a given dose of radiation was approximately 2-3 times greater than the number generated by in vivo irradiation. These results suggest that measurement of MN in vivo after irradiation can be used as an in vivo dosimeter. However, precise dosimetry is probably affected by factors such as kinetic changes in different lymphocyte populations and possibly by in vivo factors which influence sensitivity of cells to radiation.     

唐古特大黄多糖组分1对急性电离辐射损伤小鼠的保护作用

目的：探讨唐古特大黄多糖组分1（RTP1）对急性电离辐射损伤小鼠的保护作用。方法：采用昆明种小鼠,随机分为5组：正常对照组（Normal Control,NC）、辐射对照组（Irradiation Control,IC）以及RTP1低剂量组（200 mg/kg）、中（400 mg/kg）和高剂量组（800 mg/kg）,采用灌胃给药方式,连续14 d,NC组和IC组则给予等量的生理盐水,第14 d除NC组外,各组小鼠均接受2.0 Gy/只60Coγ射线照射1次,照射后24 h,检测小鼠胸腺和脾脏指数,测定肝脏超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性及丙二酰二醛（MDA）水平以及小鼠外周血象和骨髓嗜多染红细胞（PCE）微核数。结果：RTP1能够升高小鼠的胸腺、脾脏指数,增加肝脏SOD和GSH-Px活性,降低MDA水平,升高外周血中白细胞数并降低骨髓PCE微核数,与IC组比较有统计学意义（P〈0.05或P〈0.01）。结论：RTP1对辐射所致的小鼠损伤具有一定的保护作用。     

The effect of myelopid on antibody formation in gamma-irradiated animals

The effect of a new immunocorrecting preparation, Myelopid, on the antibody-forming cell content of mouse spleen after gamma-irradiation (1-3 Gy) has been investigated. The preparation administered after immunization of mice with sheep erythrocytes increases the number of antibody-forming cells in the spleen, the effect being a function of radiation dose and time interval between the exposure and immunization. The preparation is ineffective when delivered after irradiation, but prior to immunization.     

Mesenchymal stromal cells but not cardiac fibroblasts exert beneficial systemic immunomodulatory effects in experimental myocarditis

Systemic application of mesenchymal stromal cells (MSCs) in inflammatory cardiomyopathy exerts cardiobeneficial effects. The mode of action is unclear since a sufficient and long-acting cardiac homing of MSCs is unlikely. We therefore investigated the regulation of the immune response in coxsackievirus B3 (CVB3)-induced acute myocarditis after intravenous application of MSCs. Wildtype mice were infected with CVB3 and treated with either PBS, human MSCs or human cardiac fibroblasts intravenously 1 day after infection. Seven days after infection, MSCs could be detected in the spleen, heart, pancreas, liver, lung and kidney, whereby the highest presence was observed in the lung. MSCs increased significantly the myocardial expression of HGF and decreased the expression of the proinflammatory cytokines TNFα, IL1β and IL6 as well as the severity of myocarditis and ameliorated the left ventricular dysfunction measured by conductance catheter. MSCs upregulated the production of IFNγ in CD4+ and CD8+ cells, the number of IL10-producing regulatory T cells and the apoptosis rate of T cells in the spleen. An increased number of CD4+CD25+FoxP3 could be found in the spleen as well as in the circulation. In contrast, application of human cardiac fibroblasts had no effect on the severity of myocarditis and the systemic immune response observed after MSCs-administration. In conclusion, modulation of the immune response in extracardiac organs is associated with cardiobeneficial effects in experimental inflammatory cardiomyopathy after systemic application of MSCs.     

Low-dose total-body γ irradiation modulates immune response to acute proton radiation

Health risks due to exposure to low-dose/low-dose-rate radiation alone or when combined with acute irradiation are not yet clearly defined. This study quantified the effects of protracted exposure to low-dose/low-dose-rate γ rays with and without acute exposure to protons on the response of immune and other cell populations. C57BL/6 mice were irradiated with ??Co (0.05 Gy at 0.025 cGy/h); subsets were subsequently exposed to high-dose/high-dose-rate proton radiation (250 MeV; 2 or 3 Gy at 0.5 Gy/min). Analyses were performed at 4 and 17 days postexposure. Spleen and thymus masses relative to body mass were decreased on day 4 after proton irradiation with or without pre-exposure to γ rays; by day 17, however, the decrease was attenuated by the priming dose. Proton dose-dependent decreases, either with or without pre-exposure to γ rays, occurred in white blood cell, lymphocyte and granulocyte counts in blood but not in spleen. A similar pattern was found for lymphocyte subpopulations, including CD3+ T, CD19+ B, CD4+ T, CD8+ T and NK1.1+ natural killer (NK) cells. Spontaneous DNA synthesis by leukocytes after proton irradiation was high in blood on day 4 and high in spleen on day 17; priming with γ radiation attenuated the effect of 3 Gy in both body compartments. Some differences were also noted among groups in erythrocyte and thrombocyte characteristics. Analysis of splenocytes activated with anti-CD3/anti-CD28 antibodies showed changes in T-helper 1 (Th1) and Th2 cytokines. Overall, the data demonstrate that pre-exposure of an intact mammal to low-dose/low-dose-rate γ rays can attenuate the response to acute exposure to proton radiation with respect to at least some cell populations.     

Contrasting feature in the repopulation of host-type T cells in the spleens of F1----P and P----F1 radiation bone marrow chimeras

The regeneration and persistence of host- and donor-derived T cells were examined in the thymus as well as the spleen of mouse radiation bone marrow chimeras of two semiallogeneic combinations (F1----P, P----F1) with different Thy-1 markers on T cells of donor and host origins. An unexpectedly large number of host-type T cells were recovered from the spleens of F1----P chimeras, amounting to as high as 45 and 25% of total T cells at 6 and 14 weeks after bone marrow transplantation (BMT), respectively. To the contrary, the residual host-type T cells in the spleens of P----F1 chimeras disappeared quickly, resulting in less than 0.1% of total T cells at 6 weeks after BMT. It was also revealed that the number of host-type T cells in the spleens of F1----P chimeras decreased in proportion to increase of radiation dose given to the recipients.     

Efficiency of polysaccharide translam for early treatment of acute radiation illness

Antiradiation therapeutic efficiency of translam (1-->3; 1-->6-beta-D-glucan) produced by enzymatic synthesis out of laminarin, polysaccharide of Laminaria cychorioides, has been studied in four animal species (mice, guinea-pigs, dogs, monkeys). A stable curative effect has been observed following its administration within first 24 h after radiation exposure at doses that cause acute radiation sickness (about LD90). The preparation is nontoxic and has a broad therapeutic range which permits its practical application.     

Radioprotective properties of indralin combined with cystamine and mexamine

The radioprotective properties of indralin when it is used in combination with cystamine and mexamine are studied in inbred mice and rats. The mice and rats are irradiated with γ rays emitted by ⁶⁰Co at doses of 9.0 and 9.5 Gy, respectively. A combined parenteral administration of indralin and cystamine in mice at doses of 25 mg/kg each is revealed to potentiate the radioprotective properties of indralin up to a level close to the ED₅₀ effect, while the separate application of these drugs in doses of 25 mg/kg each has no or a very weak radioprotective effect. Indralin (50 mg/kg) and mexamine (12 mg/kg) injected intraperitoneally in rats are found to almost completely eliminate the animal mortality caused by gastrointestinal acute radiation syndrome; the mortality in the control radiation group reaches 60% on the seventh day after the animals have been exposed to radiation at a dose of 9.5 Gy. However, if bone-marrow acute radiation syndrome develops under the above condition of super-lethal dose, the radioprotectors have a low radioprotective effect. Under the this condition, the combined application of indralin and mexamine in the same doses has 50% of radioprotective effect reached by applying these radioprotectors separately in double doses.     

Repair processes of hemopoiesis after applying cyclophosphamide. II. Quantitative changes in basic proteins and DNA in the bone marrow, spleen, and thymus

The effect of cyclophosphamide on the organ weight, DNA and basic protein content in nuclei of bone marrow, thymus and spleen of rats was studied. Animals were examined 6, 12 and 24 hours and 3, 5, 10, 20 and 30 days after intraperitoneal application of cyclophosphamide in the doses of 100 and 200 mg X kg-1 of body weight. From the 5th day after application a marked decrease in weight and cellularity of all organs was observed. Amounts of DNA and basic proteins in nuclei of bone marrow and spleen were higher as compared with controls and their increase occurred prior to recovery of cellularity and organ weight, that was initiated within 5 and 10 days after cyclophosphamide treatment. Changes in the thymus persisted for a longer period of time and recovery was incomplete even at the day 30 after cyclophosphamide application. In accordance with these data no increase in DNA and basic protein amounts in the thymus nuclei was observed.     

Organ-specific gene expressions in C57BL/6 mice after exposure to low-dose radiation

The possibility that radiation-induced alterations in gene expression are tissue specific and are related to apoptosis was examined using samples from brain, heart, lung, spleen and intestine from female C57BL6 mice after exposure to 0.2 Gy radiation. Apoptosis was the highest in spleen and intestine, moderate in lung, and absent in brain and heart. However, the mRNA expression of Trp53 and Cdkn1a (p21) after irradiation was not different among the organ types, and immunohistochemistry revealed that all the organs expressed these two proteins after irradiation. When expression patterns of 23 genes in the organs were examined by RT-PCR, neogenine, Apo1, nuclease sensitive element binding protein 1, syntaxin, cyclin G1, hNOP56, paraoxonase and glutathione peroxidase were overexpressed after irradiation in all the organs sampled, suggesting them as universal exposure markers for low-dose radiation. Sialyltransferase may be a candidate for radiation detection in spleen and intestine, which are radiosensitive organs. Because Sod1 (Cu/ZnSOD) and alphaB crystalline were expressed only in spleen, and protein tyrosine kinase and platelet membrane glycoprotein lib were expressed in both spleen and lung, these genes may also be potential markers for detection of radiation exposure, especially low-dose radiation, in these tissues. These data suggested possible tissue-specific markers of low-dose radiation exposure and suggested potential novel genetic modifiers of radiation response.     

"Ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism

In this article is presented the result of the experiments on mice-hybrids F1(CBA x C57B1/6), which indicates the presence of the reaction of "ischemia/reperfusion" for stem cells of two "critical" cell renewal systems of organism (bone marrow and intestinal epithelium) during the irradiation under the conditions of hypoxic radioprotector application. The additional injection of the source of nitric oxide radicals-sodum nitroprusside (SNT) to the mice right after the irradiation under the conditions of hypoxic protection by serotonin, resulted the substantial increase of the survival rate of hematopoietic stem cells (registered by the methods of endogenous and exogenous colony forming in spleen) and stem cells of intestinal epithelium (registered by the method of intestinal "microcolonies"). The similar radioprotective effect was also registered during the test of survival rate of mice under tests of "bone marrow" and "intestinal" forms of radiation lethality that is evidence of the importance of the realization of phenomenon "ischemia/reperfusion" in the reaction of whole organism on the acute radiation injury. As SNP weakens the manifestation apoptosis and necrosis through competition with active forms of oxygen (AFO) during the period of "reperfusion" on basis of the found out phenomenon experimental model for studying mechanisms of stem cells damage in vivo induced by AFO and for the search of the modifiers weakening or strengthening such damage can be developed.     

Spleen and thymus lymphoid tissue in hypoxia (a biometric study)

The spleen and thymus have been studied macro- and microscopically in rats (180-200 g body mass) on the 1st, 3d, 5th, 7th, 14th and 28th days of adaptation to a decreased atmospheric pressure in the altitude chamber corresponding to lifting to 5,000 and 7,500 m (after a preliminary gradual acclimatization) and on the 14th, 28th, 42d, 56th days of readaptation. A relative mass of the organs, the white pulp section area--the transversal section area of the spleen ratio, the summation section area of its lymph nodules have been estimated. In the thymus the cortico-medullary index (CMI) has been defined. A relative mass of the spleen increases during the first week of hypoxia, and during adaptation period it somewhat decreases and stabilizes, remaining higher than in the control. At the altitude of 5,000 m the cross section area of the lymph nodules decreases by 17% on the 28th adaptation day and at the altitude of 7,500 m--by 27% beginning from the 14th up to the 28th adaptation days. In the thymus the CMI, after some decrease during the first days of hypoxia at the altitude of 5,000 m, increases and normalizes on the 28th adaptation day, and at the altitude of 7,500 m stabilizes on the 14th - 28th days of hypoxia. When the rats are at the altitudes of 5,000 and 7,500 m the thymus lymphoid tissue reacts more quickly to the hypoxia effect and much sooner normalizes during the readaptation period than does the white pulp of the spleen. The main changes in the lymphoid tissue of the spleen and thymus take place on the 7th - 28th days of hypoxia.     

Pentadecapeptide BPC 157 positively affects both non-steroidal anti-inflammatory agent-induced gastrointestinal lesions and adjuvant arthritis in rats

Besides a superior protection of the pentadecapeptide BPC 157 (an essential fragment of an organoprotective gastric juice peptide BPC) against different gastrointestinal and liver lesions, an acute anti-inflammatory and analgetic activity was also noted. Consequently, its effect on chronic inflammation lesions, such as adjuvant arthritis, and non-steroidal anti-inflammatory agents (NSAIAs)-induced gastrointestinal lesions was simultaneously studied in rats. In gastrointestinal lesions (indomethacin (30 mg/kg sc), aspirin (400 mg/kg ig) and diclofenac (125 mg/kg ip) studies, BPC 157 (10 μg or 10 ng/kg ip) was regularly given simultaneously and/or 1 h prior to drug application (indomethacin). In the adjuvant arthritis (tail-application of 0.2 mL of Freund's adjuvant) studies (14 days, 30 days, 1 year) BPC 157 (10 μg or 10 ng/kg ip), it was given as a single application (at 1 h either before or following the application of Freund's adjuvant) or in a once daily regimen (0–14th day, 14–30th day, 14th day–1 year). Given with the investigated NSAIAs, BPC 157 consistently reduced the otherwise prominent lesions in the stomach of the control rats, as well as the lesions in the small intestine in the indomethacin groups. In the adjuvant arthritis studies, the lesion's development seems to be considerably reduced after single pentadecapeptide medication, and even more attenuated in rats daily treated with BPC 157. As a therapy of already established adjuvant arthritis, its salutary effect consistently appeared already after 2 weeks of medication and it could be clearly seen also after 1 year of application. Taking together all these results, the data likely point to a special anti-inflammatory and mucosal integrity protective effect.     

Multivariate statistical analysis of tissue basophils in acute radiation sickness

By means of mark scale estimation method, discriminant and factor analyses, changes in integral indices on the state of mast cells in the rat mesentery have been investigated in dynamics of medullary, intestinal and cerebral forms of an acute radiation sickness. The integral indices are calculated basing on the morphometric parameters of the cells, that are obtained after the histological preparations are treated in a special automatic system for analysing images. At the medullary form of the acute radiation sickness, the greatest structural rearrangements of the mast cells take place during the first hours after the radiation, as well as during the climax of the disease, at the intestinal form, the analogous changes are revealed 1-3 days after the effect, and at the cerebral form--3 h after the radiation. The integral indices, calculated by means of the three methods, are well correlated with each other.     

Molecular signals in antigen presentation. II. Activation of cytolytic cells in vitro after ultraviolet radiation or combined gamma and ultraviolet radiation treatment of antigen-presenting cells

Murine low-density spleen cells have potent antigen-presenting ability in a hapten-specific cytolytic T lymphocyte (CTL) system using the hapten azobenzenearsonate (ABA). Exposure of these cells to 0.33 KJ/m2 of ultraviolet radiation (UVR) after coupling to hapten results in markedly inhibited antigen-presenting function that can be substantially corrected or bypassed by interleukin 1 (IL 1). These results have been interpreted to reflect an inhibition of Lyt-1+ T cell activation by UVR-treated APC. Treatment of these cells sequentially with 1500 rad of gamma-radiation (GR) prior to hapten coupling, followed by 0.33 KJ/m2 of UVR radiation after coupling, results in an antigen-presenting defect only minimally improved by IL 1. However, partially purified interleukin 2 (IL 2) can completely bypass or correct this defect. Thus, combined GR and UVR induces a different or more profound defect in APC function when compared to UVR alone. However, these cells do provide a signal(s) other than hapten necessary for CTL activation because ABA-coupled high density spleen cells do not activate CTL cells, even with the addition of IL 2. Fluorescence-activated cell sorter analysis demonstrates that exposure of these low density spleen cells to GR or UVR results in decreased I-A antigen expression at 24 hr than either alone. The addition of nonhapten-coupled low-density APC partially reconstitutes the ability of combined GR/UVR-treated LD-APC to present antigen, and this effect is enhanced by the administration of exogenous IL 1. This occurs despite a lack of significant accessory cell activity by the LD-APC for the ABA hapten, and indicates that combined GR/UVR-treatment of APC is not functionally equivalent to completely removing them.     

The effect of internal exposure to 90Sr on hematopoietic stem cells in CBA mice

After acute intake of 90Sr the changes of d-9 CFUs number in mice (CBA) bone marrow, spleen and peripheral blood were investigated. The obtained results indicated similar quantitative changes in bone marrow and spleen CFUs on exposure to the 90Sr when radiation doses did not cause the decrease in life-time (1.11 kBq/g). Sarcomogeneous doses of 90Sr (29.6 kBq/g) resulted in drastic changes of hemopoietic system: spleen haematopoiesis activation and suppression of bone marrow functions. On the first day after 90Sr injection (29.6 kBq/g) the increase in number of peripheral blood CFUs (circulating pool) was observed.