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Ionizing radiation-induced mouse micronucleus in peripheral blood reticulocytes dose - response relationship

Author: ZhangLing
Tutor: LiuLiBo
School: Jilin University
Course: Radiation Medicine
Keywords: Ionizing radiation Biological dosimeter Reticulocyte Micronucleus Flow cytometry Apoptosis
CLC: R144
Type: Master's thesis
Year: 2011
Downloads: 73
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Abstract

With the rapid development of nuclear science and technology, ionizing radiation technology has been in medicine, industry, agriculture, energy, geology and other fields has been widely used. Ionizing radiation to human beings in enormous economic benefits at the same time, due to various reasons will cause radiation damage to humans, or even catastrophic consequences. Early quickly and accurately estimate the personnel exposure doses of radiation injury in the early diagnosis, treatment and prognosis as well as the assessment of late effects is essential. Therefore, looking for a quick, accurate, high-throughput biological radiation dosimeter radiation injury has become the focus of research questions. Purpose of ICR mice for the study, the use of flow cytometry different dose rates of mice after X-ray irradiation of micronuclei in peripheral blood reticulocytes dose-effect relationship. Detected under the same conditions of different X-ray irradiation dose rate of mice after bone marrow cell micronucleus dose-response relationship, both as verified by comparing the experimental results objectivity and authenticity. Of micronuclei in peripheral blood reticulocytes as radiation biological dosimeter applications provide an experimental basis. Simultaneous detection of different dose rates 2GyX irradiated mice bone marrow nucleated erythroid cell apoptosis, apoptosis rate from initial inquiry micronucleus formation mechanism. Method 1. Different dose rate X-ray irradiated mice micronuclei in peripheral blood reticulocytes detection of 96 ICR mice by radiation dose rates were randomly divided into four groups, namely 0.5Gy/min group, 1.OGy/min group, 2.OGy/min group and 3.5Gy/min groups, each further divided into 0 Gy, 1.OGy, 2.OGy, 3.OGy, 4.0 Gy dose and 5.OGy six points, each dose 4 mice. Single body X-ray irradiation after 24h, the eye at peripheral blood, blood samples were fixed at -70 ℃ low temperature refrigerator for more than 24h, CD71 antibody were added to the solution, RNAase A and PI solution, using a FACScan flow cytometer collect 20,000 reticulocyte erythrocyte micronucleus assay performed. (2) X-ray irradiation at different dose rates of mice after bone marrow cell micronucleus test the packet ICR mice after blood in the eye killed by cervical dislocation, whichever bilateral femur marrow smears, fixed with methanol, Giemsa staining, counted under a microscope 2000 bone marrow polychromatic erythrocytes micronucleus rate and calculate the micronucleus rate expressed in parts per thousand. 3 different dose rates 2GyX irradiated mice bone marrow erythroid nucleated cell apoptosis detection 20 ICR mice by radiation dose rates were randomly divided into five groups, OGy / min group, 0.5Gy/min group, 1Gy/min group, 2Gy/min group, 3.5Gy/min groups. The first group as a control group, after four dose groups were 2Gy. After irradiation 24h, dislocated sacrificed syringe 5ml cold PBS buffer flushing bilateral bone marrow in mice, 300 mesh filter omentum into single cell suspension was prepared after centrifugation. Adjusting the cell concentration of 1.0 × 106 cells / ml or so, take the cell solution were added 1m1 CD71 antibody, Buffdr buffer, Hochest33342 solution and PI was added. Using a FACScan flow cytometer collect 10,000 CD71-positive cells to detect apoptosis. Results 1. Different dose rate X-ray irradiated mice micronuclei in peripheral blood reticulocytes dose-effect relationship 0-5Gy dose range of different X-ray dose rate irradiation micronuclei in peripheral blood reticulocytes rates are increasing dose is raised, and can be fitted to the linear regression equation, but the difference between the slope of the dose rate groups are different. Statistical analysis showed that: 0.5Gy/min group 1.OGy/min and 2.0Gy/min regression coefficient comparison between groups with statistically significant difference (t0.5Gy/min vs IGy / min = 6.881, P lt; 0.05; t0.5Gy/min vs 2Gymin = 4.767.P lt; 0.05); 1.0Gy/min group 2.0Gy/min and 3.5Gy/min regression coefficient comparison between groups with statistically significant difference (tlGy / min vs 2Gymin = 2.612, P lt; 0.05; tlGy / min vs 3.5Gy/min 4.896, P lt; 0.05); 2.0Gy/min group 3.5Gy/min regression coefficient comparison between groups with statistically significant difference (t2Gymin vs 3.5Gy / min = 2.756, P lt; 0.05); 0.5Gy/min group 3.5Gy/min regression coefficient comparison between groups was not statistically different. (2) X-ray irradiation at different dose rates of mice after bone marrow cell micronucleus dose-response relationship in the dose range of 0-5Gy different X-ray dose rate irradiation bone marrow cell micronucleus rates are increasing dose is raised, and can be fitted to the linear regression equation, but the difference between the slope of the dose rate groups are different. Statistical analysis showed that: 0.5Gy/min group and 1.OGy/min group, 2. OGy / min group and the comparison between the regression coefficients 3.5Gy/min groups have statistically significant difference (t0.5Gv/min vslGv / min = 7.63, P lt; 0.05; t0.5Gy/min vs 2Gymin = 6.47, P lt; 0.05: tO.5Gy/min vs 3.5Gy/min = 2.51, P lt; 0.05); 1.0Gy/min group and 2.0Gy / min and 3.5Gy/min regression coefficient comparison between groups with statistically significant difference (t1MGy/min vs 2Gv/min = 2.32, P lt; 0.05; t, Gy / min vs 3.5Gy/min = 5.05.P lt; 0.05); 2.0Gy/min group 3.5Gy/min regression coefficient comparison between groups with statistically significant difference (t2Gymin vs 3.5Gy/min = 3.56.P lt; 0.05). 3.FCM detection of micronuclei in peripheral blood reticulocytes and artificial microscopic bone marrow cell micronucleus relationship between FCM and artificial microscopy results of two methods showed a good dose-response relationship, but slightly higher than the results by FCM artificial microscopy results, statistical analysis showed that both methods detected no statistically significant difference between the results (t0.5Gy/min = 1.724.P gt; 0.05; t1.0Gy/min = 2.157, P gt; 0.05; t2. 0Gy/min = 1.288.P gt; 0.05: t3.5Gy/min = 1.333.P gt; 0.05), and both have a good correlation between (r0.05Gy/min = 0.987, P lt; 0.05; r1 .0.Gy/min = 0.993.P lt; 0.05; r2.0Gy/min = 0.992, P lt; 0.0S: r3.5Gy/min = 0.989.P lt; 0.05), 4. different irradiation dose rate 2GyX After bone marrow erythroid nucleated cells apoptosis rate in mice receiving different dose rates 2Gy X-ray irradiation, bone marrow nucleated erythroid cells was significantly higher than the control group with a statistically significant difference (P lt; 0.05), and with the increase of radiation dose rate, with each exposure group were statistically significant difference (P lt; 0.05). Conclusion 1 in the 0 ~ 5Gy dose range of different X-ray irradiation dose rate in peripheral blood reticulocytes with micronuclei increased with increasing radiation dose, the dose-response relationship can be fitted to the linear regression equation, but different doses rate between the two groups of different slopes. 2 In the dose range of 0 ~ 5Gy different X-ray irradiation dose rate of bone marrow polychromatic erythrocytes with micronuclei increased with increasing radiation dose, dose-response relationships can be fitted to the linear regression equation, but different dose rates slope between different groups. 3.FCM with two detection methods artificial microscopy showed no difference between the two and has a good correlation. Flow cytometry of peripheral blood reticulocytes rapid detection of micronuclei as biological radiation dosimeter, but prior to clinical application, requires further study. 4 apoptosis and micronucleus formation in cells to ionizing radiation damage are two forms of reaction, both exist simultaneously, a lower dose rate irradiation in micronucleus formation-based, high-dose-rate irradiation with apoptosis-based .

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