【作者】 肖勤；

【导师】 王维娜； 王安利；

【作者基本信息】 河北大学 ， 水生生物学， 2003， 硕士

【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、Na₂SeO₃浓度达到150mg／L、NH₄Cl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NH₄Cl胁迫下超氧阴离子含量（O₂^-）高于对照组，超氧化物歧化酶（SOD）活力也高于对照组（NH₄Cl浓度为400mg／L的SOD除外）；Vc组血细胞在NH₄Cl胁迫下超氧阴离子含量（O₂^-）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（NH₄Cl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NH₄Cl胁迫下超氧阴离子含量低于相应的对照组（NH₄Cl浓度为400mg／L的O₂^-含量除外），SOD活力高于相应的对照组（NH₄Cl浓度为400mg／L的SOD除外）；Se组血细胞在NH₄Cl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NH₄Cl肋迫下超氧阴离子含量低于相应的对照组（NH₄Cl浓度为400mg／L的O₂^-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。更多还原

【Abstract】 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 thancontrol 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 .更多还原