超氧化物歧化酶对荷瘤体免疫细胞的辐射防护作用
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摘要
目的：研究超氧化物歧化酶(SOD)对Lewis肺癌荷瘤小鼠受照射后淋
巴细胞脂质过氧化的影响，以及对肺癌患者外周血受照后脂质过氧化酶
活性的作用，旨在阐明SOD对淋巴细胞脂质过氧化系统的辐射抗性及其
对放疗引起的免疫功能降低的防护作用。
材料和方法：建立实验肿瘤动物模型，将Lewis肺癌皮下荷瘤小鼠随
机分为三组：对照组：照射组：SOD+照射组。后两组用医用直线加速器，
6 MeV X线对小鼠进行全身照射，照射结束后取脾脏，制备成脾细胞匀
浆，应用752型分光光度仪测定荷瘤小鼠脾细胞OD值并计算丙二醛
(MDA)、总SOD(T-SOD)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶
(GSH-PX)、过氧化氢酶(CAT)含量。取病理确诊的未经过治疗的小
细胞肺癌患者的空腹抗凝静脉血标本，每人份6ml，分为3等份，分为3
个实验组：对照组，照射组，SOD+照射组。后两组用医用直线加速器
6MVX线对血标本进行照射，照射后用752型分光光度仪测定肺癌患者
外周血淋巴细胞OD值及丙二醛(MDA)、总-SOD(T-SOD)、谷胱甘肽
(GSH)含量。
结果用SPSS10.0软件行单因素方差分析处理。
结果：荷瘤鼠照射组与对照组相比，照射后脾细胞MDA明显增加
(p＜0.05)，两组比较差异显著；而照射后荷瘤鼠脾脏中T-SOD，GSH，
GSH-PX，CAT均下降(p＜0.01)，两组比较差异显著。SOD+照射组与照
超氧化物歧化酶对荷瘤体免疫细胞的辐射防护作用中文摘要
射组比较，SOD+照射组荷瘤鼠脾脏组织匀浆中MDA含量明显下降，而
T一SOD，GSH，GSH一pX，CAT等均增加，均有显著差异(p<0.05)。
肺癌患者照射组与对照组相比，照射后外周血中Ml)A增加(P<0.05)，
T一SOD，GSH下降(p<0.05)，两组比较均有显著差异。soD+照射组与
照射组比较，soD+照射组血清中MDA含量明显下降(p<0.05)，而
T一SOD，GsH无显著变化(p>0.05)。
结论:1.电离辐射可以引起荷瘤小鼠脾脏GSH一PX，GsH，CAf等抗氧化
酶活性下降，自由基中间产物MDA增加;SOD增加受照射荷瘤小鼠脾
脏中GSH一PX，GSH，CAT等抗氧化酶活性，减少MDA含量，有明显的辐
射防护作用。
2.电离辐射可以引起肺癌患者外周血T一SOD活力和GSH含量明
显下降，MDA含量显著增加，而SOD能降低MDA含量，对受照射人
静脉血中T一SOD活力和GSH含量略有增加，但尚无统计学意义。提示
SOD可减轻电离辐射引起脂质过氧化反应，增强机体对辐射损伤的耐受
性，有明显的辐射防护作用。
Objective: to research superoxide dismutase (SOD) action on lymphocyte lipid excessive oxidation of mice bearing Lewis pulmonary carcinoma after radiation , and the action on human lymphocyte number, proportion and lipid excessive oxidation. Try to clarify SOD radiaton resistance for lymphocyte and the protection on imrmmological function reduction after radiation.
Material and method: 3 random groups of mice bearing Lewis pulmonary carcinoma include contrast group, radiation group, SOD + radiation group. The second one and the third one received total radiation, 6MeV X ray,5Gy by medical linear accelerator. The third one were injected SOD into abdominal cavity 30 minutes before radiation. After radiation, immediately draw the spleen. 752-type spectro-light degree meter detect OD value of the mice spleen lymphocyte, then we calculate MDA, T-SOD, GSH, GSH-PX, CAT in the spleen. Untreated small cell lung cancer patients diagnosticated by pathology were collected venous anticoagulation blood. The blood samples were separated 3 shares as the similar mice groups, received similar treatment and detection as mice samples. Result: Compared with the unradiation control group, the mice control group MDA in spleen rise obviously (p<0.05), but all the T-SOD, GSH,
GSH-PX, CAT values reduce (p<0.01). The SOD + radiation group MDA value reduce, T-SOD, GSH, GSH-PX, CAT values rise (p<0.05), compared with radiation group. The radiation group MDA value rise T-SOD, GSH, GSH-PX, CAT values reduce (p<0.05), compared with control group. The SOD + radiation group MDA value reduce (p<0.05), T-SOD, GSH, GSH-PX, CAT values similar (p>0.05), compared with radiation control group. Experiment in vitro tell us that radiation make T-SOD, GSH in human serum reduced, MDA rose. And SOD makes an adverse result. SOD can relieve lipid peroxidation by radiation, reinforce body tolerance to radiation.
Conclusion: 1. Radiation can reduce the mice spleen antioxidase, GSH-PX,GSH,CAT, rise free radical media products MDA. SOD product adverse result, can protect the harm from radiation. 2. Radiation can reduce T-SOD activity and GSH value in lung cancer blood, while rise MDA value. SOD product adverse result, can protect the harm from radiation.
巴细胞脂质过氧化的影响，以及对肺癌患者外周血受照后脂质过氧化酶
活性的作用，旨在阐明SOD对淋巴细胞脂质过氧化系统的辐射抗性及其
对放疗引起的免疫功能降低的防护作用。
材料和方法：建立实验肿瘤动物模型，将Lewis肺癌皮下荷瘤小鼠随
机分为三组：对照组：照射组：SOD+照射组。后两组用医用直线加速器，
6 MeV X线对小鼠进行全身照射，照射结束后取脾脏，制备成脾细胞匀
浆，应用752型分光光度仪测定荷瘤小鼠脾细胞OD值并计算丙二醛
(MDA)、总SOD(T-SOD)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶
(GSH-PX)、过氧化氢酶(CAT)含量。取病理确诊的未经过治疗的小
细胞肺癌患者的空腹抗凝静脉血标本，每人份6ml，分为3等份，分为3
个实验组：对照组，照射组，SOD+照射组。后两组用医用直线加速器
6MVX线对血标本进行照射，照射后用752型分光光度仪测定肺癌患者
外周血淋巴细胞OD值及丙二醛(MDA)、总-SOD(T-SOD)、谷胱甘肽
(GSH)含量。
结果用SPSS10.0软件行单因素方差分析处理。
结果：荷瘤鼠照射组与对照组相比，照射后脾细胞MDA明显增加
(p＜0.05)，两组比较差异显著；而照射后荷瘤鼠脾脏中T-SOD，GSH，
GSH-PX，CAT均下降(p＜0.01)，两组比较差异显著。SOD+照射组与照
超氧化物歧化酶对荷瘤体免疫细胞的辐射防护作用中文摘要
射组比较，SOD+照射组荷瘤鼠脾脏组织匀浆中MDA含量明显下降，而
T一SOD，GSH，GSH一pX，CAT等均增加，均有显著差异(p<0.05)。
肺癌患者照射组与对照组相比，照射后外周血中Ml)A增加(P<0.05)，
T一SOD，GSH下降(p<0.05)，两组比较均有显著差异。soD+照射组与
照射组比较，soD+照射组血清中MDA含量明显下降(p<0.05)，而
T一SOD，GsH无显著变化(p>0.05)。
结论:1.电离辐射可以引起荷瘤小鼠脾脏GSH一PX，GsH，CAf等抗氧化
酶活性下降，自由基中间产物MDA增加;SOD增加受照射荷瘤小鼠脾
脏中GSH一PX，GSH，CAT等抗氧化酶活性，减少MDA含量，有明显的辐
射防护作用。
2.电离辐射可以引起肺癌患者外周血T一SOD活力和GSH含量明
显下降，MDA含量显著增加，而SOD能降低MDA含量，对受照射人
静脉血中T一SOD活力和GSH含量略有增加，但尚无统计学意义。提示
SOD可减轻电离辐射引起脂质过氧化反应，增强机体对辐射损伤的耐受
性，有明显的辐射防护作用。
Objective: to research superoxide dismutase (SOD) action on lymphocyte lipid excessive oxidation of mice bearing Lewis pulmonary carcinoma after radiation , and the action on human lymphocyte number, proportion and lipid excessive oxidation. Try to clarify SOD radiaton resistance for lymphocyte and the protection on imrmmological function reduction after radiation.
Material and method: 3 random groups of mice bearing Lewis pulmonary carcinoma include contrast group, radiation group, SOD + radiation group. The second one and the third one received total radiation, 6MeV X ray,5Gy by medical linear accelerator. The third one were injected SOD into abdominal cavity 30 minutes before radiation. After radiation, immediately draw the spleen. 752-type spectro-light degree meter detect OD value of the mice spleen lymphocyte, then we calculate MDA, T-SOD, GSH, GSH-PX, CAT in the spleen. Untreated small cell lung cancer patients diagnosticated by pathology were collected venous anticoagulation blood. The blood samples were separated 3 shares as the similar mice groups, received similar treatment and detection as mice samples. Result: Compared with the unradiation control group, the mice control group MDA in spleen rise obviously (p<0.05), but all the T-SOD, GSH,
GSH-PX, CAT values reduce (p<0.01). The SOD + radiation group MDA value reduce, T-SOD, GSH, GSH-PX, CAT values rise (p<0.05), compared with radiation group. The radiation group MDA value rise T-SOD, GSH, GSH-PX, CAT values reduce (p<0.05), compared with control group. The SOD + radiation group MDA value reduce (p<0.05), T-SOD, GSH, GSH-PX, CAT values similar (p>0.05), compared with radiation control group. Experiment in vitro tell us that radiation make T-SOD, GSH in human serum reduced, MDA rose. And SOD makes an adverse result. SOD can relieve lipid peroxidation by radiation, reinforce body tolerance to radiation.
Conclusion: 1. Radiation can reduce the mice spleen antioxidase, GSH-PX,GSH,CAT, rise free radical media products MDA. SOD product adverse result, can protect the harm from radiation. 2. Radiation can reduce T-SOD activity and GSH value in lung cancer blood, while rise MDA value. SOD product adverse result, can protect the harm from radiation.
引文
1、谷铣之，刘泰福，潘国英主编。肿瘤放射治疗学[M]。人民卫生出版社。190-233．
2、陈瑗，脂过氧化物与辐射损伤．中华放射医学与防护杂志，1984，4：63-67。
3、方允中，李文杰．自由基与酶 第一版，北京：科学出版社，1987：251-254。
4、Halliwell B, Gulteridge JMC.Oxygen free radicals and iron in relation to biology and medicine; some problems and concepts.Arch. Biochem. Biophys, 1986,246:501-514.
5、刘树铮，医学放射生物学[M]．北京原子能出版社，1986，20-22。
6、Kazuhiro Nomura, Hirotaka Imai, Tomoko Koumura,et,al. Mitochondrial phospholipid hydroperoxide glutathione peroxidase suppresses apoptosis mediated by a mitochondrial death pathway.[J] The journal of Biological Chemistry. vol.274,No.41,Issue of October 8, 1999, 29294-29302.
7、Mirochnitchenko O, Inouye M.Effect of overexpression of human Cu,Zn superoxide dismutase in transgenic mice on macrophage functions. J Immunol 1996 Feb 15;156(4): 1578-86.
8、Rodgers, M.A.J. and Powers, E.L. Oxygen and Oxy-radiation in Chemistry and Biology, New York: Academic press, 1981.
9、Petkau A.Role of superoxide dismutase in modification of radiation injury. Br J Cancer Suppl 1987 Jun;8:87-95.
10、赵厚安，方允中．整体照射对小鼠肝、脾亚细胞结构各部分中锰-超氧化物歧化酶及铜锌-超氧化物歧化酶活力的影响．生物化学与生物物理学报，1989，21：195-200
11、Petkau A, Sargent MD, Chelack WS, et,al. Response of human bone marrow progenitor cells to X-rays in vitro. Int J Radiat Biol 1988 Oct;54(4):593-600
12、.Pani G, Bedogne B, Anzevino R et al. Detegulated manganese superoxide dismutase expression and resistance to oxidative injury in p53-deficident cells. Cancer Res, 2000,60(14):3927-3939
13、陈晓穗，骆训懿，周丽君等 重组人铜锌超氧化物歧化酶对小鼠抗氧化酶和免疫细胞的辐射防护作用．中华放射医学与防护杂志，1997，17(3)：189-192。
14、Finnegan MT, Herbert KE, Evans MD,et,al. Evidence for sensitisation of DNA to oxidative damage during isolation. Free Radic Biol Med 1996;20(1):93-8.
15、谷先之，刘泰福，潘国英主编．肿瘤放射治疗学[M]．北京：人民卫生出版社，1981．
16、孙存普，张建中，段绍瑾主编．自由基生物学导论[M]．合肥：中国科学技术出版社，1999．194-210．
17、毕爱华主编．医学免疫学．人民军医出版社，1995，240
18、Sweetman SF, Strain J J, McKelvey-Martin VJ.Effect of antioxidant vitamin supplementation on DNA damage and repair in human lymphoblastoid cells. Nutr Cancer 1997;27(2): 122-30
19、郑荣梁主编．自由基生命科学进展[M]．北京：原子能出版社，2001．39-43．
20、Takahashi H, Hashimoto Y, Aoki N, et al. Copper, zinc-superoxide dismutase protects from ultraviolet B-induced apoptosis of SV40-transformde human keratinocytes:the protection is associated with the increased levels of antioxidant enzymes. J Dematol Sci, 2000,23:12-21
21、Prarer A, Terry NHA, Murray D. Influence of intracellular thiol and polyamine levels on radioprotection by aminorhiols. Int J Radiat Biol, 1993,64:71-81
22、Park DS, Morris EJ, Stefanis L,et al. Multiple pathways of neuronal death induced by DNA-damaging agents,NGF deprivation,and oxidative stress. J Neurosci, 1998,18;830-840
23、Ushakova T, Melkonyan H, Nikonova L, et al. Modification of gene expression by dietary antioxidants in radiation-induced papooses of mice splenocytes. Free Radic Biol Med, 1999,26:887-891
24、Mcbrien DCH,Slater TF. Free Radiation, Lipid Peroxidation and Cancer (London) 1982;317.
2、陈瑗，脂过氧化物与辐射损伤．中华放射医学与防护杂志，1984，4：63-67。
3、方允中，李文杰．自由基与酶 第一版，北京：科学出版社，1987：251-254。
4、Halliwell B, Gulteridge JMC.Oxygen free radicals and iron in relation to biology and medicine; some problems and concepts.Arch. Biochem. Biophys, 1986,246:501-514.
5、刘树铮，医学放射生物学[M]．北京原子能出版社，1986，20-22。
6、Kazuhiro Nomura, Hirotaka Imai, Tomoko Koumura,et,al. Mitochondrial phospholipid hydroperoxide glutathione peroxidase suppresses apoptosis mediated by a mitochondrial death pathway.[J] The journal of Biological Chemistry. vol.274,No.41,Issue of October 8, 1999, 29294-29302.
7、Mirochnitchenko O, Inouye M.Effect of overexpression of human Cu,Zn superoxide dismutase in transgenic mice on macrophage functions. J Immunol 1996 Feb 15;156(4): 1578-86.
8、Rodgers, M.A.J. and Powers, E.L. Oxygen and Oxy-radiation in Chemistry and Biology, New York: Academic press, 1981.
9、Petkau A.Role of superoxide dismutase in modification of radiation injury. Br J Cancer Suppl 1987 Jun;8:87-95.
10、赵厚安，方允中．整体照射对小鼠肝、脾亚细胞结构各部分中锰-超氧化物歧化酶及铜锌-超氧化物歧化酶活力的影响．生物化学与生物物理学报，1989，21：195-200
11、Petkau A, Sargent MD, Chelack WS, et,al. Response of human bone marrow progenitor cells to X-rays in vitro. Int J Radiat Biol 1988 Oct;54(4):593-600
12、.Pani G, Bedogne B, Anzevino R et al. Detegulated manganese superoxide dismutase expression and resistance to oxidative injury in p53-deficident cells. Cancer Res, 2000,60(14):3927-3939
13、陈晓穗，骆训懿，周丽君等 重组人铜锌超氧化物歧化酶对小鼠抗氧化酶和免疫细胞的辐射防护作用．中华放射医学与防护杂志，1997，17(3)：189-192。
14、Finnegan MT, Herbert KE, Evans MD,et,al. Evidence for sensitisation of DNA to oxidative damage during isolation. Free Radic Biol Med 1996;20(1):93-8.
15、谷先之，刘泰福，潘国英主编．肿瘤放射治疗学[M]．北京：人民卫生出版社，1981．
16、孙存普，张建中，段绍瑾主编．自由基生物学导论[M]．合肥：中国科学技术出版社，1999．194-210．
17、毕爱华主编．医学免疫学．人民军医出版社，1995，240
18、Sweetman SF, Strain J J, McKelvey-Martin VJ.Effect of antioxidant vitamin supplementation on DNA damage and repair in human lymphoblastoid cells. Nutr Cancer 1997;27(2): 122-30
19、郑荣梁主编．自由基生命科学进展[M]．北京：原子能出版社，2001．39-43．
20、Takahashi H, Hashimoto Y, Aoki N, et al. Copper, zinc-superoxide dismutase protects from ultraviolet B-induced apoptosis of SV40-transformde human keratinocytes:the protection is associated with the increased levels of antioxidant enzymes. J Dematol Sci, 2000,23:12-21
21、Prarer A, Terry NHA, Murray D. Influence of intracellular thiol and polyamine levels on radioprotection by aminorhiols. Int J Radiat Biol, 1993,64:71-81
22、Park DS, Morris EJ, Stefanis L,et al. Multiple pathways of neuronal death induced by DNA-damaging agents,NGF deprivation,and oxidative stress. J Neurosci, 1998,18;830-840
23、Ushakova T, Melkonyan H, Nikonova L, et al. Modification of gene expression by dietary antioxidants in radiation-induced papooses of mice splenocytes. Free Radic Biol Med, 1999,26:887-891
24、Mcbrien DCH,Slater TF. Free Radiation, Lipid Peroxidation and Cancer (London) 1982;317.