OBJECTIVETo investigate the effects of microwave radiation on thymocytes in mice at different power densities.

METHODSThe experimental animals were whole-body exposed to microwave radiation with frequency of 2,450 MHz, power density of 1, 5, 15 mW/cm(2) respectively 1 h everyday for 30 days. Then the thymus were taken out after the mice were decapitated. Thymus index, morphological characteristics of thymus were examined. The changes of thymus T-cell subgroups, cell cycle progression in thymocytes and cellular apoptosis were detected with flow cytometry (FCM).

RESULTSThe body weights of animals in 5, 15 mW/cm(2) irradiation groups [(28.10 +/- 1.46), (27.50 +/- 2.52) g] were lower than that of the control [(31.95 +/- 2.51) g] (P < 0.05). Pathological observation showed dark red piece of nucleus, some nuclei inclined to one side, slight increase in hassall body. The expressions of CD8 in 5, 15 mW/cm(2) irradiation groups (29.14% +/- 1.68%, 29.18% +/- 0.81%) were higher than that in control group (26.95% +/- 1.27%) (P < 0.05). The percentages of G(2) + M phase thymocytes in both radiation groups (12.24% +/- 1.82%, 11.19% +/- 1.36%) were lower than that in control group (14.58% +/- 0.64%) (P < 0.01). Thymocytic apoptosis rates in the three experimental groups (7.18% +/- 0.99%, 10.06% +/- 1.58%, 9.45% +/- 0.92%) were higher than that in control (4.25% +/- 1.63%) (P < 0.01), but the evident difference between 5 mW/cm(2) and 15 mW/cm(2) was not found (P > 0.05).

CONCLUSIONSub-chronic microwave exposure (2 450 MHz, 5, 15 mW/cm(2)) could induce thymocyte apoptosis, cause pathological changes in thymus, and affect cell cycle progression, thus may inhibit the immune function of the animal.

Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; Female ; Male ; Mice ; Microwaves ; adverse effects ; T-Lymphocytes ; radiation effects ; Thymus Gland ; cytology ; radiation effects

OBJECTIVETo investigate the role of apoptosis in radiation-induced mouse thymus lymphocyte damage and repair and provide the basis for understanding the molecular mechanism of radiation-induced lymphocyte damage and repair as well as the prevention and treatment of acute radiation sickness.

METHODSWe studied the dynamic changes of apoptosis of mouse thymus lymphocytes and the expression of bax and bcl-2 gene products after 2, 4, 6 and 8 Gy of whole body gamma-irradiation using in situ terminal labeling, DNA electrophoresis and immunohistochemical techniques.

RESULTSAt the early stage after irradiation, the percentage of apoptotic lymphocytes increased rapidly in accordance with the increasing of radiation doses, while the counts of the thymus and peripheral lymphocytes decreased sharply, showing an opposite change to lymphocyte apoptosis. After 6 Gy gamma-irradiation, typical morphological characteristics of thymus apoptotic lymphocytes in early, middle and late stages were found by transmission electron microscopy. The thymus lymphocytes displayed characteristic DNA ladders 4 hr and 8 hr after 2-6 Gy gamma-irradiation,using DNA gel electrophoresis techniques. Abnormal expression of bcl-2 and bax gene products were shown in irradiated lymphocytes.

CONCLUSIONSApoptosis plays an important role in the process of radiation-induced mouse thymus lymphocyte damage and repair. Bcl-2 and Bax proteins may regulate the process of lymphocyte apoptosis.

Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; Gamma Rays ; Lymphocytes ; physiology ; radiation effects ; Male ; Mice ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Thymus Gland ; pathology ; radiation effects ; Time Factors ; bcl-2-Associated X Protein

We previously determined that AKR/J mice housed in a low-dose-rate (LDR) (137Cs, 0.7 mGy/h, 2.1 Gy) gamma-irradiation facility developed less spontaneous thymic lymphoma and survived longer than those receiving sham or high-dose-rate (HDR) (137Cs, 0.8 Gy/min, 4.5 Gy) radiation. Interestingly, histopathological analysis showed a mild lymphomagenesis in the thymus of LDR-irradiated mice. Therefore, in this study, we investigated whether LDR irradiation could trigger the expression of thymic genes involved in the DNA repair process of AKR/J mice. The enrichment analysis of Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways showed immune response, nucleosome organization, and the peroxisome proliferator-activated receptors signaling pathway in LDR-irradiated mice. Our microarray analysis and quantitative polymerase chain reaction data demonstrated that mRNA levels of Lig4 and RRM2 were specifically elevated in AKR/J mice at 130 days after the start of LDR irradiation. Furthermore, transcriptional levels of H2AX and ATM, proteins known to recruit DNA repair factors, were also shown to be upregulated. These data suggest that LDR irradiation could trigger specific induction of DNA repair-associated genes in an attempt to repair damaged DNA during tumor progression, which in turn contributed to the decreased incidence of lymphoma and increased survival. Overall, we identified specific DNA repair genes in LDR-irradiated AKR/J mice.
Animals ; DNA Repair/*radiation effects ; Dose-Response Relationship, Radiation ; Female ; Gene Expression Regulation/*radiation effects ; Gene Regulatory Networks/radiation effects ; Lymphoma/etiology/*genetics ; Mice ; Mice, Inbred AKR ; Oligonucleotide Array Sequence Analysis ; *Radiation, Ionizing ; Reverse Transcriptase Polymerase Chain Reaction ; Thymus Gland/*radiation effects ; Thymus Neoplasms/etiology/*genetics

This study was purposed to investigate the effect of G-CSF on the proliferation, differentiation, and cell cycle distribution of thymocytes in sublethally irradiated mice. Female BALB/c mice were exposed to 6.0 Gy γ-ray irradiation and then randomly divided into control and G-CSF treatment group. In the treatment group rhG-CSF 100 µg/(kg·d) was given subcutaneously for 14 continuous days and to make sure the first injection was given within 1 hour after irradiation. Cell cycle distribution and apoptosis of thymocytes were detected within 72 hours after irradiation. Subpopulations of CD4(-)CD8(-) cells and sequential changes in the distribution of CD4(+)CD8(+), CD8(+)CD4(-), CD8(-)CD4(+) cells were detected by a three-color flow cytometry during a four-weeks period after irradiation. The results showed that in G-CSF treatment group marked increase of cells in G(0)/G(1) phase (G-CSF vs control: 82.0 ± 5.0% vs 75.9 ± 2.8%) (p < 0.05) and a decrease of cells in S phase (G-CSF vs control: 10.2 ± 4.8% vs 15.7 ± 2.3%) (p < 0.05)could be observed as early as 6 hours after irradiation, but G-CSF seems have no evident effects on the cells in G(2)/M phase. G-CSF could also protect thymocytes against apoptosis. 6 and 12 hours after irradiation the apoptosis rates of thymic cells in G-CSF treatment group were 11.5 ± 2.4% and 15.5 ± 3.3% respectively, while in the control group the apoptosis rates were 16.5 ± 2.2% and 22.6 ± 0.7% respectively. Comparison between the two group demonstrated significant difference (p < 0.05). CD4(-)CD8(-) double negative thymocytes (DN)can be defined as DN1-4 according to their maturation. G-CSF treatment resulted in a significant increase in DN1 thymocytes and promoted their proliferation and differentiation to a more mature DN3 and DN4 stage. G-CSF could enhance the recovery of CD4(+)CD8(+) thymocytes and mitigate their relapse during reconstitution. The percentage of CD4(+)CD8(+) thymocytes in the G-CSF treatment group 28 days after irradiation was significantly higher than that of the control group (71.0 ± 6.3% vs 25.5 ± 6.3%) (p < 0.05). It is concluded that G-CSF has a positive effects on the thymic cell cycle distribution, proliferation and differentiation, which may contribute to the reconstitution of central immune system after acute irradiation.
Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; pharmacology ; therapeutic use ; Lymphocyte Count ; Mice ; Mice, Inbred BALB C ; Radiation Injuries, Experimental ; therapy ; Thymus Gland ; cytology

OBJECTIVETo observe the effects of signal factors of corticosterone (CS), cAMP, cGMP, Ca2+ andprotein kinase C (PKC) on lymphocyte apoptosis in mouse thymus induced by X-rays of 4 Gy in vitro.

METHODSThe DNA lytic rate for thymocytes was measured by fluorospectrophotometry.

RESULTSThe DNA lyric rate for thymocytes 4-8 hours after irradiation with 2-8 Gy was significantly higher than that in the control (P<0.01). As compared with the control, the DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.05-0.4 microg/mL ionomycin (Iono, P<0.05 or P<0.01) or 0.05-0.4 ng/mL phorbol myristate acetate (PMA, P<0.05 or P<0.01), respectively, was significantly increased, while the rate for thymocytes treated with 50 ng/mL cGMP was not significantly increased. The DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.2 and 0.4 microg/mL Iono (P<0.05), and 0.2 and 0.4 ng/mL PMA (P<0.05) plus 4-Gy irradiation, respectively, was significantly higher than that treated with single 4-Gy irradiation, while the rate for thymocytes treated with 50 ng/mL cGMP plus 4-Gy irradiation was not increased. When both 0.4 microg/mL Iono and 0.4 ng/mL PMA acted on the thymocytes, the DNA lytic rate for thymocytes was significantly higher than that in the control (P<0.01), the DNA lytic rate for thymocytes treated with both 0.4 microg/mL Iono and 0.4 ng/mL PMA plus 4-Gy irradiation was significantly higher than that treated with single 4-Gy irradiation (P<0.05), but was not significantly higher than that treated with 0.4 microg/mL Iono plus 4-Gy irradiation or 0.4 ng/mL PMA plus 4-Gy irradiation.

CONCLUSIONCS, cAMP, Ca2+, and PKC signal factors can promote thymocyte apoptosis induced by larger dose X-rays.

Animals ; Apoptosis ; drug effects ; radiation effects ; Calcium ; pharmacology ; Corticosterone ; pharmacology ; Cyclic AMP ; pharmacology ; Cyclic GMP ; pharmacology ; Ionomycin ; pharmacology ; Male ; Mice ; Protein Kinase C ; metabolism ; Spectrometry, Fluorescence ; Tetradecanoylphorbol Acetate ; pharmacology ; Thymus Gland ; cytology ; drug effects ; X-Rays

OBJECTIVETo investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes.

METHODSFluorescent staining and flow cytometry analysis were employed for the measurement of protein expression.

RESULTSIn time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P < 0.05, P < 0.01, and P < 0.05, respectively) and at 24 h for splenocytes (P < 0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P < 0.05-P < 0.01) and from 8 h to 72 h for splenocytes (P < 0.05-P < 0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P < 0.05-P < 0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P < 0.05) and 0.5-6.0 Gy for splenocytes (P < 0.05-P < 0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure.

CONCLUSIONThe results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays.

Animals ; Cyclin D1 ; biosynthesis ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; Cyclin-Dependent Kinases ; biosynthesis ; Dose-Response Relationship, Radiation ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Proto-Oncogene Proteins ; Radiation Dosage ; Spleen ; cytology ; metabolism ; radiation effects ; Thymus Gland ; cytology ; metabolism ; radiation effects ; X-Rays

PURPOSE: Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidyl choline to phosphatidic acid (PA) and choline. Recently, PLD has been drawing much attentions and considered to be associated with cancer process since it is involved in cellular signal transduction. In this experiment, oleate-PLD activities were measured in various tissues of the living rats after whole body irradiation. MATERIAL AND METHODS: The reaction mixture for the PLD assay contained 0.1microCi 1,2-di[1-14C]palmitoyl phosphatidylcholine, 0.5mM phosphatidylcholine, 5mM sodium oleate, 0.2% taurodeoxycholate, 50mM HEPES buffer (pH 6.5), 10mM CaCl2, and 25mM KF. phosphatidic acid, the reaction product, was separated by TLC and its radioactivity was measured with a scintillation counter. The whole body irradiation was given to the female Wistar rats via Cobalt 60 Teletherapy with field size of 10cm x 10cm and an exposure of 2.7Gy per minute to the total doses of 10Gy and 25Gy. RESULTS: Among the tissues examined, PLD activity in lung was the highest one and was followed by kidney, skeletal muscle, brain, spleen, bone marrow, thymus, and liver. Upon irradiation, alteration of PLD activity was observed in thymus, spleen, lung, and bone marrow. Especially PLD activities of the spleen and thymus revealed the highest sensitivity toward gamma-ray with more than two times amplification in their activities. In contrast, the PLD activity of bone marrow appears to be reduced to nearly 30%. Irradiation effect was hardly detected in liver which showed the lowest PLD activity. CONCLUSION: The PLD activities affected most sensitively by the whole-body irradiation seem to be associated with organs involved in immunity and hematopoiesis. This observation strongly indicates that the PLD is closely related to the physiological function of these organs. Furthermore, radiation stress could offer an important means to explore the phenomena covering from cell proliferation to cell death on these organs.
Animals ; Attention ; Bone Marrow ; Brain ; Cell Death ; Cell Proliferation ; Choline ; Cobalt ; Female ; Hematopoiesis ; HEPES ; Humans ; Hydrolysis ; Kidney ; Liver ; Lung ; Muscle, Skeletal ; Oleic Acid ; Phosphatidic Acids ; Phosphatidylcholines ; Phospholipase D* ; Phospholipases* ; Radiation Effects ; Radioactivity ; Rats* ; Rats, Wistar ; Scintillation Counting ; Signal Transduction ; Sodium ; Spleen ; Taurodeoxycholic Acid ; Thymus Gland ; Whole-Body Irradiation