抗氧化物对日本沼虾离体细胞的影响
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
本文以日本沼虾为实验材料,首先在M199基础培养基中加入不同浓度的抗氧化物(维生素C、维生素E、硒)及氨氮对日本沼虾肌肉细胞进行传代培养,利用溴化四唑蓝(MTT)法检测传代细胞增殖活力,得出:当Vc浓度达到100mg/L、VE浓度达到200mg/L、Na_2SeO_3浓度达到150mg/L、NH_4Cl浓度为150mg/L时,它们的增殖活力最高。然后选取其增殖活力最高的抗氧化物浓度加入到培养基中培养日本沼虾的肌肉细胞与血细胞,在氨氮胁迫后测定离体细胞抗氧化酶的变化,结果如下:(1)维生素C对氨氮胁迫下日本沼虾离体细胞的影响表明:Vc组肌肉细胞在NH_4Cl胁迫下超氧阴离子含量(O_2~-)高于对照组,超氧化物歧化酶(SOD)活力也高于对照组(NH_4Cl浓度为400mg/L的SOD除外);Vc组血细胞在NH_4Cl胁迫下超氧阴离子含量(O_2~-)高于相应的对照组,而超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GPX)活力却低于相应的对照组,过氧化氢酶(CAT)活力高于对照组(NH_4Cl浓度为400mg/L的CAT除外)。(2)硒(以下简称Se)对日本沼虾离体细胞的影响表明:日本沼虾离体肌肉细胞SOD活力随着Se浓度增大而升高,当达到150mg/L时酶活力最高并随后开始下降,这与不同浓度的硒对细胞增殖活力的影响一致;Se组肌肉细胞在NH_4Cl胁迫下超氧阴离子含量低于相应的对照组(NH_4Cl浓度为400mg/L的O_2~-含量除外),SOD活力高于相应的对照组(NH_4Cl浓度为400mg/L的SOD除外);Se组血细胞在NH_4Cl胁迫下SOD活力低于相应的对照组,而GPX活力高于相应的对照组。(3)维生素E对氨氮胁迫下日本沼虾离体细胞的影响表明:VE组肌肉细胞在NH_4Cl肋迫下超氧阴离子含量低于相应的对照组(NH_4Cl浓度为400mg/L的O_2~-含量除外),而SOD活力高于相应的对照组;VE组血细胞在
    
    摘要
    NH4CI胁迫下超氧阴离子含量、SOD活力均高于相应的对照组。(4)维生素E
    与硒(以下简称VESe)对氨氮胁迫下日本沼虾离体细胞的影响表明:VESe组
    叭肉细胞在NH4CI胁迫一「超氧阴离子含量高于对照组,SOD活力也高于对照组
    (NH;el浓度为40Omg/L的SOD除外);VESe组血细胞在NH4CI胁迫下SOD
    与GPX活力均高于相应的对照组。
This paper presents results obtained on macrobrachium nipponense muscle cell and haemocytes in primary culture.
    To begin with, when vitamin C is 100mg/L, vitamin E is 200mg/L, Na2SeO3 is 150mg/L, NH4Cl is 150mg/L in M199 medium separately, muscle cell proliferation activity is the highest in subculture. The next, when muscle cell and haemocytes in primary culture are exposed to high ammonia-N, the production of superoxide anions (O2-) and the activities of antioxidant enzymes are caused to change by vitamin C, vitamin E, Na2SeO3 separately. The results showed as follow: (1) Vc group are higher than control group in the production of superoxide anions(O2-) and the activity of SOD of primary culture muscle cells ; Vc group is higher than control group in the production of superoxide anions(O2-) of primary culture haemocytes. but Vc group are lower than control group in the activities of SOD and GPX of primary culture haemocytes. Vc group is higher than control group in the activity of CAT. (2) Se group is lower than control group in the production of superoxide anions (02") of primary culture muscle cells. But Se group
    is higher than control group in the activity of SOD; Se group is lower than control group in the activity of SOD of primary culture haemocytes. But Se group is higher than control group in the activity of GPX of primary culture haemocytes; In addition, the activity of SOD is positive relation to the concentration of Na2SeO3 in primary culture muscle cells (3) VE group is lower than
    
    
    
    
    control group in the production of superoxide anions (O2-) of primary culture muscle cells. But Se group is higher than control group in the activity of SOD; VE group are higher than control group in the production of superoxide anions (O2-) and the activity of SOD of primary culture haemocytes. (4) VESe group are higher than control group in the production of superoxide anions (O2-) and the activity of SOD of primary culture muscle cells; VESe group are higher than control group in the activities of SOD and GPX of primary culture haemocytes .
引文
[1] 孙建贻,张道源,段中华,谭德清.洪湖日本沼虾的种群繁殖生物学.湖泊科学,1998,10(2):72-77.
    [2] 胡国宏、朱蕊成、张俊辉.日本沼虾的人工繁殖与养殖试验.内陆水产,2000,25(12):31-35.
    [3] 王立新,杨朝霞.动物细胞培养及应用.黄牛杂志,2000,26(3):45-48.
    [4] 司徒镇强,吴军正.细胞培养.世界图书出版公司,1996,1-5.
    [5] 张晓华,王立平,徐怀恕.对虾组织培养研究进展及其开发应用的潜在价值.海洋湖沼通报,1996,23(2):78-82.
    [6] Chen SN,Kou GH.Infection of cultured cells from the lymphoid organ of Penaeus monodon Fabricus by monodon-type baculovirus(MBV).J.Fish Dis., 1989,12:73-76.
    [7] Chert SN,Shin HH,Kou GH. Primary cell cultures from tissuesof Penaeid shrimps and their susceptibilities to monodon-type baculovirus(MBV).Reports on Fish Disease Research, 1997,16:1-14.
    [8] Chen SN,Shih HH,Kou GH. Primary cell cultures from tissues of penaeid shrimps and their susceptibilities to Monodon-type Baculovirus(MBV).COA Fish Ser., 1995,53:1-14.
    [9] Chen SN,Wang CS. Establishment of cell culture system from penaeid shrimp and their susceptibility to white spot disease and yellow head viruses.Methods Cell Sci., 1999,21(4):199-206.
    [10] Fuerst JA,Sambhi SK,Paynter JL, et al. Isolation of a bacterium resembling Pirellula spp. From primary tissue culture of the giant tiger prawn(Penaeus monodon).Applied and Environmental Microbiology, 1991,57(11):3127-3134.
    [11] Itami T, Maeda M,Kondo M et al. Primary culture of lymphoid organ cells and haemocytes of kuruma shrimp,Penaeus japonicus.Methods Cell Sci.,21(4):237-244.
    [12] Lu YA, Nadala EC, Brock JA, Loh PC,1991.A new virus isolate from infectious hypodermal and hematopoietic necrosis virus(IHHNV) infected penaeid shrimps.J.Virol.Methods, 1999,31 (2/3):189-195.
    [13] Lu Y, Tapay LM,Loh PC et al.Development of a quantal assay in primary shrimp cell culture for yellow head baculovirus(YBV) of penaeid shrimp.Journal of Virological Methods,52(1-2):231-236.
    [14] Tapay LM,Lu Y, Gose RB et al,Development of an in vitro quantal assay in primary cell cultures for a non-occluded baculo-like virus of penaeid shrimp.Journal of Virological Methods,64(1):37-41.
    [15] 苗宏志,童裳亮,徐斌,姜明,刘晓云.利用对虾原代细胞增殖对虾杆状病毒HHNBV的研
    
    究.生物工程学报,2000,16(2):221-224.
    [16] 苗宏志,童裳亮,徐斌,姜明,刘晓云.中国对虾淋巴组织培养中的病毒及病理观察.水产学报,1999,23(2):169-173.
    [17] 汪岷,姜明,樊廷俊,刘晓云,汪晓峰,徐怀恕.中国对虾淋巴组织培养细胞中立克次氏体增殖的形态学研究.海洋科学,200,24(3):3-5.
    [18] 汪岷,樊廷俊,姜明,王馗,李红岩,徐怀恕.中国对虾(Penaeus chinensis)淋巴组织培养细胞中一种球形病毒的超微结构的研究.青岛海洋大学学报,2000,30(2):111-114.
    [19] Fraser CA,Hall MR. Studies on primary cell cultures derived from ovarian tissue of Penaeus monodon.Methods Cell Sci., 1999,21(4):213-218.
    [20] 李霞.浅谈虾、贝类细胞培养方法和应用前景.国外水产,1993,4:1-3.
    [21] Hose J E and Martin G G. Defense functions of granulocytes in ridgeback prawn Sicyonia ingentis.J. Invertebrate Patho., 1989,53:335-346.
    [22] 薛庆善.体外培养的原理与技术.科学出版社,2001,8-16.
    [23] 王秀玲,董苍转.细胞培养在毒理学研究中的应用.国外医学卫生学分册.2000,27(3):178-181.
    [24] 张天宝.睾丸细胞体外培养方法及在生殖毒理研究中的应用.卫生毒理学杂志,1995,9(2):121-123.
    [25] 蔡生力,杨丛海.17α-羟孕酮对离体培养的对虾卵巢组织发育的促进作用.海洋水产研究,2000,21(2):7-10.
    [26] 李爱杰.维生素C对中国对虾的营养和免疫作用及其最适剂型.粮食与饲料工业.1998,2:33-36.
    [27] Yamamoto,Y. et al.Biochemical studies on L-ascorbic acid in aquatic animals,Bull.Jap.Soc.Sci.Fish, 1977,43(1):53-57.
    [28] Wolf K,Quimby M C.Established eurythemic line of fish cells in vitro.Science, 1962,135:1065-1066.
    [29] 徐志吕,刘铁斌,雷清新,李爱杰.中国对虾(Penaeus chinensis)Ⅴ.维生素营养的研究中国对虾维生素C营养.青岛海洋大学学报,1994,24(3):364-377.
    [30] Guary, M.,Kanazawa,A.,Tanaka,N..Requirement for ascorbic acid Nutritional requirements of prawn. Mam.Fac. Fish. KagoshinaUniv.,1976,25:53-57.
    [31] Magarelli,RC.,Hunter, B.,Lightner, D.V..Black death, an ascorbic acid deficiency disease in Peraeid shrimp.Comp. Biochem.Physiol., 1979,63A: 103-108.
    [32] 王安利,等.中国对虾配合饵料中维生素C添加量的研究.91全国海水养殖学术讨论会论文集,1992,380-386.
    [33] 周洪琪,邱小琮,华雪铭.水产动物的营养与免疫、抗病能力.健康养殖专栏
    [34] 王伟庆等.抗坏血酸聚磷脂盐对中国对虾生长、缺氧耐受力及免疫抵抗力的影响.海洋湖沼通报,1996,1,42-49.
    [35] 谢平,许毅.微量元素锌硒与免疫.微量元素与健康研究,1998,15:68.
    
    
    [36] Zikic R V, Stajn A S, Ognjanovic BI,et al.The effect of cadmium and selenium on the antioxidant enzyme activities in ratheart.J Environ Pathol Toxicol Oncol,1998,17(3-4):259.
    [37] 张叔人,张友会,陆燕蓉.硒对巨噬细胞功能的调变.中华微生物和免疫学杂志,1988,8(2):73.
    [38] 王新刚,邬惠琼,柴莲花.硒多糖与亚硒酸钠的抗突变性研究.同济医科大学学报,1997,26(2):89.
    [39] 刘国廉,李煜,樊飞跃,等.微量元素硒的防癌抗癌作用.中国药理学与毒理学杂志,1997,11(2):28.
    [40] 惠天朝,施明华,朱荫湄.硒对罗非鱼慢性镉中毒肝抗氧化酶及转氨酶的影响.中国兽医学报,2000,20(1):264-266.
    [41] Combs GF, Noguchi T, Scott ML. Machanisms of action of selenium and vitamin E in protection of biological membranes[J].Fed Proc., 1975,34:2090.
    [42] 谢巧雄,朱心玲.亚硒酸钠对四氯化碳损伤鱼肝原代细胞与肝组织的保护作用[J].水生生物学报.1996,20(3):229-235.
    [43] 呙于明.家禽营养与饲料.北京中国农业大学出版社,1997,119.
    [44] 邹仕庚.猪与鸡的维生素E营养.饲料研究,1998,6:14-17.
    [45] Ray WM and Chien YH.Effects of stocking density and aged sediment on tiger prawn Penaeus monodon, nursery system.Aquaculture, 1992,104(3-4):231-248.
    [46] Chen JC and Kou YZ.Effects of ammonia on growth and molting of penaeus japonicus.A quaculture, 1992,104:249-260.
    [47] Chen JC and Kou YZ.Accumulation of ammonia in the hemolymph penaeus japonicus exposed to ambient ammonia.Dis.Aquat.Org., 1991,11: 187-191.
    [48] Chen JC,Cheng SY andChen CT.Changes of haemocyanin,protein and free aminoacid levels in the haemolymph of penaeus japonicus exposed to ambient ammonia.Comp.Biochem. Physiol., 1994,109A:339-347.
    [49] Young-Lai WW, Charmantier-Daures M,Chharmantier G. Effect of ammonia on survival and osmoregulation in different life stages of the lobster homarus americanus.Mar. Biol.,1991, 110:293-300.
    [50] Chen JC.Study on the free amino acid levelsin the hemolymph,gill,hepatopancreas and muscle of Penaeus monodon exposed to elevated ambient ammonia. Aquatic.Toxicol.,2000,50(1-2):27-37.
    [51] Chen JC,Cheng SY. Hemolymph PCO2,hemocyanin,protein levels and urea excretions of Penaeus monodon exposed to ambient ammonia. Aquatic.Toxicol., 1993,27(3-4):281-292.
    [52] Chen JC, Lin CY. Responses of oxygen consumption, Ammonia-Nexcretion and Urea-N excretion of Penaeus chinensis exposed to ambient ammonia at different salinity and pH levels.Aquaculture, 1995,136(3-4)243-255.
    [53] Chen JC, Nan FH. Effect of ambient ammonia on ammonia-N excretion and ATPase
    
    activity of Penaeus chinensis. Aquat.Toxic.,1992,23:1-10.
    [54] 孙舰军,丁美丽.氨氮对中国对虾抗病力的影响.海洋与湖沼.1999,30(3):267-272.
    [55] 方允中,李文杰.1994.自由基与酶.1994,科学技术出版社.
    [56] 浅田浩二.生物体内的活性氧清除系统—抗氧化酶.日本医学介绍,1994,15(1):293-294.
    [57] 黎婉玲,阎书凤.氧自由基与超氧化物歧化酶的文献综合利用(-).广州医药,1998,29(1):4-7.
    [58] Thomas CE and Aust SD. Rat liver microsomal NADPH-dependent release of iron from ferritin and lipid peroxidantion. Free Radical Biol.Med., 1985,1:293-300.
    [59] 魏静,陆承平,黄捷,杨丛海.克氏原螯虾的血淋巴细胞原代培养.畜牧与兽医,1999,31(5):11-12.
    [60] 陈志阳,王国华,杨敏等.一种简便快速检测淋巴细胞增殖的方法.中国生化药物杂志,2000,21(5):245-247.
    [61] 林忠宁等.MTT法检T淋巴细胞增殖功能的方法学探讨与应用.中国卫生检验杂志,2000,10(1):8-10.
    [62] Marcelo M, Ricardo C, Jenny R et al. Measurement of reactive oxygen intermediate production in haemocytes of the penaeid shrimp, Penaeus vannamei.Aquaculture, 2000,191:89-107.
    [63] 静天玉,赵晓瑜.用终止剂改进超氧化物歧化酶邻苯三酚测活法.生物化学与生物物理进展,1995,22(1):84-86.
    [64] Beers RFand Sizer IW.Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase.J.Biol.Chem., 1952,195:133-140.
    [65] 夏奕明,朱莲真.血和组织中谷胱甘肽过氧化物酶活力的测定方法.卫生研究,1987,16(4):29-33.
    [66] Mosmann T. Rapid colorimetric assay for cellar growth and survival:application proliferation and cytotoxity assays.J Immunol Methods, 1983,65(1):5.
    [67] 兰州大学生物系细胞遗传教研室.细胞生物学实验.高等教育出版社,1986,18-20.
    [68] 薛建华,刘忠权,李政年,常家文,肖存杰.维生索C和谷胱甘肽对离体肺泡巨噬细胞超微弱发光的影响.营养学报,2000,22(4):322-324.
    [69] 徐松波,张学莉,姜毅民.食品营养与健康指南[M].北京:中国医学出版社,1992,110-111.
    [70] 衰老防御自由基损害的抗衰老药物现状与进展.
    [71] Burdon RH and Rice-Evants C.Free radicals and the regulation of mammalian cell proliferation. Free Rad Res Comms.,1989,6(6):345.
    [72] 呼文亮,需静洪,张海莲.维生素E和硒对正常及其恶性细胞增殖的联合作用.延安大学学报,1995,14(1):59-62.
    [73] Chan HW-S and Coxon DT.Lipid hydroperoxides.In:Autoxidation of unsaturated lipids. London.Acadmic Press, 1987,17-50.
    [74] 杨俊诚,朱永懿,陈景坚.硒的生物学研究进展.农业环境与发展.1994,11(4):17-21.
    
    
    [75] 李勇等.硒对体外培养大鼠胚胎的发育毒性实验研究.四川环境,1994,13(4):24-27.
    [76] 卞伟,王冬武,杨红权,罗梦良.氨氮对三角帆蚌及鱼中毒死亡分析.内陆水产,1996,5:2-3.
    [77] 李丹萍.氨氮对鱼类的影响.科学养鱼,1998,5:14.
    [78] B. Fauconneau. Sensitivity of muscle satellite cells to pollutants:an in vitro and in vivo comparative approach. Aquatic Toxicology,2001,53(3-4):247-263.
    [79] Kevin Dowling and Carmel Mothersill. The further development of rainbow trout primary epithelial cell cultures as a diagnostic tool in ecotoxicology risk assessment. Aquatic Toxicology, 2001,53(3-40):279-289.
    [80] 于爱平,马景德.活性氧与细胞凋亡.前卫医药杂志,1999,1(4):254—255.
    [81] Allen R G, Balin A K, Oxidative influnce on development and differentiation: A overview of a free radical theroy of development. Free Radical Biology & Medicine,1989, 6(6):631-661.
    [82] McCord J M,Keele B B, Fridovich I.Proc.Natl.Acad.Sci.USA, 1971;68:1024.
    [83] 陈瑗,周玫.自由基医学.北京:人民军医山版社,1991:199.
    [84] Hallwell B et al. Free radical in biology and medicine. Clarendon Press.Oxford, 1989, 106-123.
    [85] 李培峰,方允中.活性氧所导致超氧化物歧化酶疏水性的变化.生物化学与生物物理进展,1995,22(1):47-50.
    [86] 李培峰,方允中.活性氧对蛋白质的损伤作用.生物化学杂志,1995,11(1):76.
    [87] Guary M,A Kanazawa,N Tanaka and HJ Ceccaldi.Nutritional requirements of prawn-Ⅵ. Requirements for ascorbic acid. Mem.Fac.Vish.,Kagoshima Univ., 1976,25:52-57.
    [88] Lightner DV, B Hunter, PC Magarelli,Jr.and LBColvin.Ascorbic acid:Nutritional requirement and role in wound repair in penaeid shrimp. Proc.World Maricul.Soc., 1979, 10:513-528.
    [89] 黄兴国,贺建华,鲁玖华,左建军.硒的营养作用及其在畜禽生产中的应用.湖南农业大学学报(自然科学版),2002,28(1):85-42.
    [90] Yiming X et al.Biochemical studies of Se- deficient population in China:Measurement of selenium,glutathione peroxidase and other oxidant defense indices in blood.Journal of Nutrition, 1989, 119:1318.
    [91] Rotruck JT.Selenium:Biochemical role as a component or glutathione peroxidase.Science, 1972, 179(2):588.
    [92] Heng Long H et al.Changes in free radicals,trace elements,and neurophysiological function in rats with liver damage induced by D-galactosamine.Biological Trace Element Research, 1992,34:19.
    [93] Johansson E et al.Selenium and its protection against the effects of mercury and silver.Journal of Trace Elements Electrolityte Health Disease, 1991,5:273.
    [94] Mills GC Hemoglobin catabolism, 1, glutethione peroxidase, an erythrocyteenzyme, which protects hemoglobin from oxidative break,J,Biol.Chen., 1957,229:189-197.
    
    
    [95] Flo é L,Gunzler W A,Schock H H.Glutathion peroxidase:a selenoenzyme.FFBS Lett., 1973,32:132-134.
    [96] 王春霞,王子健,彭安等.用自旋捕集技术研究硒化合物对活性氧自由基的消除作用.生物化学与生物物理学报,1994,26(6):585-590.
    [97] 徐辉碧等.硒化合物毒性的自由基机理.华中理工大学学报,1991,19(1):13.
    [98] 李贵明,李金有.硒对大鼠血清蛋白、血脂的亚慢毒性作用研究.中国公共卫生,1997,13(11):690.
    [99] 张欣文,王旭辉,白莉华等.亚硒酸钠对果蝇谷胱甘肽过氧化物酶活力及寿命的影响.卫生研究,2000,29(3):166-167.
    [100] 杜泽吉,苏燎原,孙国器.二甲亚砜和维生素E对照射小鼠脂质过氧化的影响.辐射研究与辐射工艺学报,1994,12(1):26-29.
    [101] Dandapat J, Chainy G B, Rao K J. Dietary vitamin E modulates antioxidant defence system in giant freshwater prawn, Macrobrachium rosenbergii.,2000,127(1):101-115.
    [102] 蔡中华,邢克智,董双林.维生素E对鲤鱼健康的影响.动物学报,2001,47(专刊):120-124.
    [103] 张士珍,王兴亚,齐志明等.硒和维生素E在硒缺乏动物自由基代谢中作用机制的研究.畜牧兽医学报,1997,28(4):311-317.
    [104] 刘丽华,呼文亮,曹守义.硒与维生素E对需谷胱甘肽抗氧化酶的保护作用.广东微量元素科学,1999,6(4):8-10.
    [105] Hu Wengliang, Goldring C E P, Rao N H et al. Effect of peroxidation and antioxidants on glutathione peroxidase activity in mammalia fibroblasts in vitro. J Xi'an Univ., 1993,5(1):22-26.
    [106] 徐瑷民等.微量元素和VE对大鼠血清脂质过氧化物和SGPT的影响.营养学报,1991,13(1):16.
    [107] Whitacre M E et al.Selenium:and mitocondrial integrity in the pancreas of the check.J Nutr., 1983,113:1972.
    [108] Combs G F et al.Mechanism of action of selenium and Vitamin E in protection of biological membrances.Fed Proc., 1975,30:2029.
    [109] Oski F A.Vitamin E:a radical defense.New England Journal of Medicine(USA),1980,303(8):454.
    [110] 王远亮.生物硒的研究进展.生物化学与生物物理进展,1987,(6):28.
    [111] Nishimoto M,Yangida GK,Stein JE,Baird WMand Varanasi U. The metabolism of benzo(a)pyrene by english sole (Parophrys vetulus):comparison between isolated hepatocytes in vitro and liver in vivo. Xenobiotica, 1992,22:949-961.
    [112] Masfaraud J-F, Devaux A, Pfohl-Leszkowicz A,Malaveille C and Monod G. DNA adduct formation and 7-ethoxyresorufin O-deethylase induction in primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene. Toxcol. In Vitro, 1992,6:523-531.
    
    
    [113] Sturm A,Cravedi JP, Perdu E et al. Effects of prochloraz and nonylphenol diethoxylate on hepatic biotransformation enzymes in trout:a comparative in vitro/in vivo-assessment using cultured hepatocytes. Aquatic Toxicology,2001,53:229-245.
    [114] Monod G,Devaux A,Valotaire Y, Cravedi JP. Primary cell cultures from fish in ecotoxicology.Fish Ecotoxicology, 1998,39-60.
    [115] Cravedi JP, Lafuente A,Baradat M,Hillenweck A,Perdu-Durand E. Biotransformation of pentachlorophenol, aniline and biphenyl in isolated rainbow trout(Oncorhynchus mykis) hepatocytes:comparison with in vivo metabolism.Xenobiotica, 1999, 29: 499-509.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700