發(fā)布時(shí)間：2018-04-08 22:31

  本文選題：α粒子　切入點(diǎn)：γ射線　出處：《復(fù)旦大學(xué)》2012年碩士論文

【摘要】：隨著核能和核科學(xué)技術(shù)的發(fā)展和廣泛應(yīng)用,電離輻射對(duì)生態(tài)環(huán)境和人類健康的影響越來越受到重視。a粒子是環(huán)境和職業(yè)暴露中的一種重要射線,除室內(nèi)氡所產(chǎn)生的低劑量α輻射外,核電站反應(yīng)堆事故所導(dǎo)致的α粒子照射將可能對(duì)公眾產(chǎn)生顯著的輻射損傷。在核事故中常常存在各種射線的聯(lián)合照射,包括不同比例的a和γ射線。另外,放射性同位素所產(chǎn)生的α射線在核醫(yī)學(xué)腫瘤治療中的應(yīng)用也日益廣泛。由于α粒子具有較高的傳能線密度(LET)和較強(qiáng)的相對(duì)生物學(xué)效應(yīng),它對(duì)人體健康的影響一直是放射生物學(xué)研究領(lǐng)域中最活躍的前沿之一。人體造血系統(tǒng)是具有較高輻射敏感性的組織之一,而微核是輻射損傷的敏感指標(biāo)。本研究采用淋巴細(xì)胞檢測(cè)分析了受a粒子、γ射線以及不同比例射線聯(lián)合照射后細(xì)胞微核率的劑量效應(yīng),檢測(cè)了兩種射線照射間隔時(shí)間對(duì)微核形成的影響。通過本研究,以期為不同類型射線聯(lián)合照射的生物劑量評(píng)價(jià)及輻射防護(hù)提供新的實(shí)驗(yàn)依據(jù)。 生物劑量計(jì)是物理劑量的有效補(bǔ)充,為了研究適用于高通量快速分析的生物劑量計(jì),本文還初步探討了蛋白分子γ-H2AX.CDKN1A作為生物劑量計(jì)的可能性。 第一部分：α粒子、丫射線聯(lián)合照射劑量效應(yīng)研究 目的探討α粒子、γ射線單獨(dú)及聯(lián)合照射引起淋巴細(xì)胞微核率的劑量效應(yīng),分析不同LET射線是否存在適應(yīng)性反應(yīng)。 方法采用人B淋巴母細(xì)胞HMy2.CIR(簡(jiǎn)稱為HMy),分別按以下條件進(jìn)行輻射：①以0.1、0.25、0.5、0.75、1、2、3、4、5Gy的γ射線照射；②以0.01、0.025、0.05、0.1、0.15、0.2、0.5、0.75和1Gy的241Ama粒子照射；為了對(duì)細(xì)胞進(jìn)行α照射,將淋巴細(xì)胞培養(yǎng)于自制的以3μm Mylar膜為底的培養(yǎng)皿中,Mylar膜預(yù)先以相對(duì)分子質(zhì)量為150-300kd的多聚賴氨酸包被過夜,使得淋巴細(xì)胞能夠貼壁生長(zhǎng)。③對(duì)于聯(lián)合照射,先采用0.025、0.1、0.2、0.5Gy的α粒子照射,然后立刻以0.25、0.75、2、3和4Gy的γ射線進(jìn)行照射；或者先采用0.25、0.75、1、2Gy的γ射線后,然后立刻以0.025、0.1、0.2、0.5Gy的α粒子進(jìn)行照射；④0.025、0.1Gy的α粒子照射后4h給予2Gy的γ射線,或0.1Gyγ射線照射后4h給予0.2、0.5a粒子照射。用細(xì)胞分裂阻滯法(CB法)檢測(cè)細(xì)胞微核發(fā)生率。結(jié)果①對(duì)γ射線照射,微核率劑量效應(yīng)符合線性平方模型Y=c+aD+βD2；②對(duì)α粒子照射,當(dāng)照射劑量小于0.25Gy時(shí),細(xì)胞微核率隨劑量的增加線性增加,當(dāng)其劑量進(jìn)一步增加時(shí),微核率劑量效應(yīng)曲線呈現(xiàn)下弓型,可以采用反映輻射旁效應(yīng)的BaD模型Y=c+αD+σ(1-exp(-δD))exp(-βD)進(jìn)行很好的擬合。③對(duì)于聯(lián)合照射,先照射α粒子時(shí),當(dāng)α粒子劑量較低時(shí),微核率的劑量效應(yīng)與Y射線照射時(shí)的相似；但當(dāng)α粒子劑量較大時(shí),微核率的劑量效應(yīng)則更接近于a粒子照射。同時(shí),0.2、0.5Gyα粒子照射聯(lián)合γ射線照射引起的微核率顯著高于單獨(dú)照射時(shí)的微核率之和(t=5.22～11.86,P0.01)。先照射γ射線時(shí),給予小劑量γ射線0.25Gy后立刻給予0.2、0.5Gy的α粒子,細(xì)胞微核率顯著高于單獨(dú)照射時(shí)的微核率之和(t=1.92,0.30,P0.01)；但當(dāng)先給予0.75、2Gy γ射線后再照射0.2Gyα粒子時(shí),微核率與單獨(dú)照射時(shí)微核率之和無顯著差別,而給予0.5Gy α粒子顯示引起的微核率高于單獨(dú)照射時(shí)的微核率之和(t=2.79,4.42,P0.05),且有統(tǒng)計(jì)學(xué)意義。一般說來,先照射α粒子的聯(lián)合照射引起的微核率高于先照射γ射線的聯(lián)合照射。④對(duì)于適應(yīng)性試驗(yàn),小劑量α粒子照射后Oh與4h給予γ射線照射,所引起的微核率無差異；而先給予0.1Gyγ射線,4h后給予α粒子照射,可以降低細(xì)胞微核率(P0.05)。 結(jié)論α粒子照射具有與γ射線不同的輻射損傷規(guī)律,輻射旁效應(yīng)可能在其中發(fā)揮了重要作用。對(duì)于先α粒子后γ射線的聯(lián)合照射可以協(xié)同增強(qiáng)輻射損傷：而先γ射線后α粒子的聯(lián)合輻射,當(dāng)α射線劑量小于0.2Gy時(shí),聯(lián)合照射表現(xiàn)為拮抗效應(yīng)；當(dāng)α射線劑量大于0.2Gy時(shí),聯(lián)合照射產(chǎn)生協(xié)同效應(yīng)。另外,當(dāng)兩次照射之間具有適當(dāng)?shù)臅r(shí)間間隔時(shí),小劑量的γ射線可誘發(fā)輻射適應(yīng)性反應(yīng),而小劑量的a粒子則未出現(xiàn)適應(yīng)性反應(yīng)。 第二部分；新型蛋白生物劑量計(jì)探討 目的探討DNA損傷修復(fù)相關(guān)蛋白γ-H2AX、CDKNIA作為生物劑量計(jì)的可能性。 方法人B淋巴母細(xì)胞HMy2.CIR(HMy)給予一定劑量α粒子或γ射線照射后,分析γ-H2AX及CDKN1A的時(shí)間響應(yīng)關(guān)系,獲知表達(dá)量較高的時(shí)間點(diǎn),并在此時(shí)間點(diǎn)上,以流式細(xì)胞儀檢測(cè)受0.01-1Gy α粒子或0.1-5Gyγ射線照射后,γ-K2A、CDKNIA蛋白表達(dá)量的劑量效應(yīng)。 結(jié)果①經(jīng)過α粒子和γ射線照射后,細(xì)胞γ-H2AX蛋白表達(dá)的第一次峰值分別出現(xiàn)在照射后0.5h和2h；對(duì)這兩種輻射,γ-H2AX蛋白在照射后24h時(shí)仍有較顯著表達(dá)；經(jīng)不同劑量α粒子、γ射線照射后,γ-H2AX蛋白劑量效應(yīng)具有一定的規(guī)律性。②經(jīng)過α粒子輻射后,細(xì)胞CDKN1A蛋白表達(dá)的第一次峰值出現(xiàn)在照射后3h,在照射后24h出現(xiàn)第二次高表達(dá)；Y射線照射后此蛋白的表達(dá)在照射后24h出現(xiàn)了增高。 結(jié)論不同LET輻射后,細(xì)胞γ-H2AX蛋白和CDKN1A蛋白表達(dá)有一定的時(shí)間響應(yīng)規(guī)律,γ-H2AX蛋白表達(dá)具有一定的劑量效應(yīng)。
[Abstract]:With the development of nuclear energy and nuclear science and technology and wide application of ionizing radiation on the ecological environment and human health has been paid more and more attention of.A particles is an important environment and occupation in the X-ray exposure, in addition to low doses of radiation generated by the indoor radon alpha, alpha particle irradiation in reactor of nuclear power plant accident caused by may the radiation produced significant damages to the public. There are often various rays combined with radiation in nuclear accident, including the different proportion of a and gamma ray. In addition, the application of alpha ray isotope produced in nuclear medicine in the treatment of cancer is increasing. The alpha particle has high linear energy transfer (LET) and strong relative biological effect, its impact on human health has been one of the most active research areas in radiation biology. The human hematopoietic system with high radiation sensitivity of a tissue, The micronucleus test is a sensitive indicator of radiation damage. This study uses lymphocytes analyzed by a particles, gamma ray dose effect and micronucleus rate of different proportion of X-ray combined with irradiation, detected the effects of two irradiation time interval on micronucleus formation. Through this study, in order to provide new experimental evidence for biological dose evaluation of different types of radiation combined with radiation and radiation protection.
Biological dosimeter is an effective supplement to physical dose. In order to study the biological dose meter suitable for high-throughput and fast analysis, we also probed into the possibility of protein molecule gamma -H2AX.CDKN1A as a biological dosimeter.
The first part: A Study on the dose effect of alpha particle and joint irradiation
Objective to investigate the dose effect of alpha particles, gamma ray alone and combined irradiation on the micronucleus rate of lymphocyte, and to analyze the adaptive response of different LET rays.
Methods using human B lymphoblastoid cell line HMy2.CIR (HMy), respectively according to the following conditions: 1. The radiation of gamma ray irradiation on 241Ama 0.1,0.25,0.5,0.75,1,2,3,4,5Gy; the 0.01,0.025,0.05,0.1,0.15,0.2,0.5,0.75 and 1Gy particles; for Zhaoshe to cell culture dish will be in lymphocyte culture based on 3 m Mylar film as the bottom of the self-made. Mylar film in advance with a relative molecular weight of 150-300kd poly-L-lysine overnight, making lymphocytes adherent. The combined irradiation, first using 0.025,0.1,0.2,0.5Gy's alpha particle irradiation, then immediately irradiated with gamma ray 0.25,0.75,2,3 and 4Gy; or by 0.25,0.75,1,2Gy gamma rays, and then immediately exposure to alpha particles of 0.025,0.1,0.2,0.5Gy; the 0.025,0.1Gy alpha particle irradiation after gamma ray 4H giving 2Gy, 0.1Gy or gamma irradiation 4h give 0.2,0.5a particle irradiation. Split block method (CB method) using cell micronucleus rate were detected. Results of gamma ray irradiation, the micronucleus rate of dose effect consistent with the linear quadratic model of Y=c+aD+ beta D2; the alpha particle irradiation, when the dose is less than 0.25Gy, the micronucleus rate increased linearly with the increase of dose, when the dose was further increased, the micronucleus rate showed a dose effect curve under the arch type, can be used to reflect the bystander effect of BaD model Y=c+ D+ (1-exp (- alpha sigma delta D)) exp (beta D) fitted very well. The first alpha particle irradiation combined with irradiation, when the dose of alpha particles when low dose Y irradiation effect and micronucleus rate is similar; but when alpha particles at high doses, dose effect on micronucleus rate is more close to the a particle irradiation. At the same time, 0.2,0.5Gy induced by alpha particle irradiation combined with gamma ray irradiation of the micronucleus rate was significantly higher than those of single irradiation The micronucleus rate and (t=5.22 - 11.86, P0.01). The first gamma rays and alpha particles giving small doses of gamma ray 0.25Gy after 0.2,0.5Gy immediately, the micronucleus rate of micronucleus rate was significantly higher than that of single irradiation and the (t=1.92,0.30, P0.01); but first give 0.75,2Gy gamma ray irradiation after 0.2Gy alpha the particle, the micronucleus rate and micronucleus rate of single irradiation and no significant difference, but given the 0.5Gy alpha particle display micronucleus rate induced micronuclei rate was higher than that of single irradiation and the (t=2.79,4.42, P0.05), and the difference was statistically significant. In general, combined with radiation caused by irradiation of alpha particle irradiation to the micronucleus rate higher than the first gamma rays. 4 for adaptability test, small dose of alpha particle irradiation after Oh and 4H treated with gamma irradiation, caused by the micronucleus rate had no difference; and give the 0.1Gy ray, 4H treated with alpha particles irradiation, can reduce the micronucleus rate (P0.05).
Conclusion alpha particle irradiation with radiation injury and regularity of different gamma ray radiation, bystander effect may play an important role in them. The first alpha particles combined with gamma rays can enhance radiation damage: first gamma ray after alpha particles combined with radiation, when alpha radiation dose is less than 0.2Gy, combined with radiation performance antagonistic effect; when the alpha dose is more than 0.2Gy, combined with radiation synergy. In addition, when has the proper time interval between the two irradiation, gamma ray radiation can induce a small dose of adaptive response, while the a particles in small doses did not show adaptive responses.
The second part of the new protein biological dosimeter
Objective to investigate the possibility of DNA damage repair related protein gamma -H2AX and CDKNIA as a biological dosimeter.
Methods human B lymphoblastoid cell line HMy2.CIR (HMy) to give a certain dose of alpha particle or gamma ray irradiation, gamma -H2AX and CDKN1A analysis of the time response relationship, get a higher expression of the time, and this time, flow cytometry was used to detect 0.01-1Gy alpha particles or 0.1-5Gy gamma ray irradiation, gamma -K2A, CDKNIA protein expression dose amount.
Results after the alpha particle and gamma ray irradiation, the first peak expression of -H2AX cells appeared 0.5h in gamma irradiation and 2H; on the two kinds of gamma radiation, there is still a significant expression of -H2AX protein in 24h after irradiation; with different doses of alpha particles, gamma irradiation, gamma dose -H2AX protein the effect has a certain regularity. After the alpha particle radiation, the first peak cell CDKN1A expression in 3h after irradiation, second high expression appeared at 24h after irradiation; Y after irradiation the expression of this protein in irradiated 24h had increased.
Conclusion after different LET radiation, the expression of gamma -H2AX protein and CDKN1A protein has a certain time response law, and the expression of gamma -H2AX protein has a certain dose effect.

【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R814.2

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