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Thesis Advisor王德铭
Degree Grantor中国科学院水生生物研究所
Place of Conferral中国科学院水生生物研究所
Degree Discipline水生生物学
Abstract本实验报导了测定ATPase活力的快速方法。对化学法则ATPase活力所用的几种测磷方法进行了比较,通过实验研究了匀浆方法、缓冲液、酶纯化步骤。对酶浓度-活力曲线、测定方法的精密度也进行了研究。用此方法测定了草鱼鳃、肾Na~+ K~+ Mg~(2+)——ATPase活力并对结果进行单因素方差分析,证实了在正常情况下,不同批鱼在温度16 ± 4 ℃驯化三天以上后,鳃、肾Na~+ K~+ Mg~(++)——ATPase活力没有显著差异(P < 0.05)。在此基础上,对Hg~(2+)、Cr~(6+),三氯联苯(TCB),直链烷基苯磺酸钠(LAS)对草鱼组织ATPase的影响进行了离体实验。结果表明:Hg~(2+), TCB, LAS对草鱼组织Na~+k~+Mg~(++)-ATPase有较强的抑制作用,浓度对数对抑制百分数的几率单位作图有较好的线性关系。它们对草鱼组织Na~+K~+Mg~(++)——ATPase的半抑制浓度分别为1.8ppm (肾), 18.44ppm (鳃), 3.92ppm (鳃)。酶实验还表明,肾Na~+K~+Mg~(++)-ATPase比鳃Na~+K~+Mg Mg~(++)——ATPase对Hg~(2+)的敏感性大,Hg~(2+),LAS对ATPase的作用有失活作用存在。Hg~(2+)主要作用于Na~+K~+-ATPase。在毒物作用水平上,对本实验的方法进行了验证。总结了毒物作用规律,讨论了ATPase实验的意义,提出了可用毒物对ATPase的影响作为毒理学实验的一项指标并对这一指标应用的可行性及前景进行了讨论。
Other AbstractA rapid method for measurement of ATPase activity was presented in this paper. Three methods of determination of inorganic phosphate be used to chemically measuring ATPase activity were compared. Homogenizing method of tissue, buffer constitutent adopted in experiment, and purification procedure of enzyme are were confirmed by experiment. The relationship between enzyme concentration and activity and the precision of this method were also studied. Gill and kidney Na~+ K~+ Mg~+ - ATPase activities of grass crap were measured by this method. And one way variance analysis was used in data handling. At normal condition, it was proved that differences of Na~+ K~+ Mg~(++)-ATPase of gill and kidney between fishes which were traped at different seasons and acclimated at certain temperature over three days were not significant (p < 0.05) Then, tests of effects of Hg~(2+), Cr~(6+), TCB (trichlo lorinated biphenyls) LAS(linear alkyl benzylsulfonate) on ATPases of grass crap tissues in vitro were carried out. The results showed that Hg~(2+), TCB and TAS had prominent inhibition on ATPase of grass crap tissues, Satisfied straight lines were obtained by pltting of percentage inhibition of ATPase on a probability axis vs. toxicants concentration on a log arthmic axis. The concentration required to yield 50% inhibition of Hg~(2+), TCB LAS to grass crap tissues were I. 8ppm(kidney), 18.44ppm(gill), and 3.92ppm(gill), respectively. The enzyme test also indicated that the sensitivity of kidney ATPase to Hg~(2+) was greater than that of gill to Hg~(2+). It was found that inactivation existed in effects of Hg~(2+), LAS on enzyme. The main action site of Hg~(2+) was Na~+ K~+ - ATPase. The method used in the experiment was verified on the level of reaction of enzyme to toxicants. The pattern of toxicants action were summarized. The significance of ATPase test was discuss It was proposed that effect of toxicants on ATPase could be used as an index of toxicological experiment. And the feasibility and perspective of this index were also discussed.
Document Type学位论文
Recommended Citation
GB/T 7714
徐立红. 用环境毒物对草鱼组织ATPase的影响作为生态毒理学指标的研究[D]. 中国科学院水生生物研究所. 中国科学院水生生物研究所,1985.
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