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Alternative TitleComparative studies on tissue distribution of microcystins in crucian carp and Japanese white rabbit
Thesis Advisor谢平
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword微囊藻毒素 鲫鱼 日本长耳兔 腹腔注射 组织分布
Abstract近年来蓝藻水华频繁发生,其伴随产生的毒素与人类健康的关系受到了越来越多的关注。微囊藻毒素(MC)是已发现的各种不同蓝藻毒素中分布最广、危害最为严重的一类肝毒素,其通过强烈地抑制蛋白磷酸酶PP1和PP2A的活性而导致机体肝损伤。本文通过急性毒性实验,采用高效液相色谱-质谱联用法(HPLC-MS)测定了MC在鲫鱼和日本长耳兔中的各个组织器官分布情况,并结合前人的研究比较了MC在鱼类和哺乳动物中组织分布的差异,为评价MC的毒性提供了依据。主要结果和结论如下: 通过腹腔注射MC提取液(200和50 μg MC-LReq./kg bw),采用LC-MS法测定了注射后1、3、12、24、48和144 h鲫鱼各组织器官中的MC含量。腹腔注射后,MC很快进入了血液循环系统并被迅速运送到鲫鱼的各个器官,表现为各组织器官不同的毒素含量水平。MC-RR在所有检测出MC的样品中占了很高的比例,而MC-LR除了在200 µg/kg剂量组鲫鱼的肝脏中相对较高(11-597 ng/g DW)之外,其它样品中均很低,甚至没有被检测出来,如肠、脾、脑、肾及肌肉等,MC-LR和MC-RR在鲫鱼体内的分布存在的差异可能是因为MC的器官特异性。在两个剂量组中,MC的最大含量(MC-RR + MC-LR)均出现在注射后1 h的血液中,分别是3757 ng/g DW (200 µg/kg剂量组)和270 ng/g DW (50 µg/kg剂量组)。两个剂量组鲫鱼肾脏中的MC-RR含量和血液中的MC-RR含量显著付相关,表明血液对于运送MC-RR进入肾脏进一步得到排泄是很重要的。腹腔注射后鲫鱼肝重/体重比在两个剂量组均增大,50 µg/kg剂量组特别明显,鲫鱼肝重/体重比增大可能是因为肝脏水肿引起的。MC在鲫鱼肝脏中清除的很快,经过48 h,96.3% (200 µg/kg剂量组)和85.7% (50 µg/kg剂量组)累积的MC被清除。鲫鱼的心脏、性腺及脑中有不同程度的MC累积,提示MC对这些器官也可能具有毒性。 MC对日本长耳兔的急性毒性实验:通过腹腔注射MC提取液(200和50 μg MC-LReq./kg bw),采用LC-MS法测定了注射后1、3、12、24和48 h日本长耳兔各组织器官中的MC含量。腹腔注射50 μg/kg剂量的MC提取液后约3 h,超过半数的兔子死亡,说明MC对兔子的毒性很强。两个剂量组的兔子血液中检测到的MC水平均较低,仅为注射剂量的0.02%-0.40%。在50 µg/kg剂量组,MC含量(MC-RR + MC-LR)在肾脏中最高(62-158 ng/g DW),其次是肝脏(54-82 ng/g DW),在12.5 µg/kg剂量组,MC含量(MC-RR + MC-LR)在肝脏中最高(15-65 ng/g DW),而且在48 h与1 h比显著升高(p < 0.01),说明肝脏中的MC难以清除。胃肠道中也检测到相对较高的MC含量,胆、脾中的MC含量没有下降的趋势,可能是由于MC在体内的再循环引起的。肾、肺及脑中有不同程度的MC累积,提示MC对这些器官也可能具有毒性。
Other AbstractWith frequent occurrence of cyanobacterial blooms in recent years, cyanotoxins have become a great threat to aquatic animals, livestock, and human health. Among these toxins, microcystins (MCs) are the most common and dangerous hepatotoxins, which intensely inhibit protein phosphatases 1 and 2A in liver, leading to hepatic damage. In this paper, acute toxicity experiments were conducted in crucian carp and Japanese white rabbit to exame tissue distribution of MCs, using liquid chromatography coupled with mass spectrometry (HPLC-MS). Combining previous studies, we provided more information on toxicity of MCs by comparing tisuuse distribution of MCs between fish and mammal. The main results and conclusions were summarized as follows: Crucian carp were injected intraperitoneally with MC extract at two doses (200 and 50 μg MC-LReq./kg bw), and MC concentrations in various tissues were analyzed at 1, 3, 12, 24, 48 and 144 h postinjection, using LC-MS. MCs entered into the circulatory system rapidly after intraperitoneal injection, and were transported to various organs, which subsequently resulted in different levels of MCs in various tissues. In all samples with detectable toxins, MC-RR was dominant wheras MC-LR contents were extremely low or even undetectable, except in the liver sample at a dose of 200 µg/kg (11-597 ng/g DW). Such differences between the distribution of MC-RR and MC-LR in fish tissues might be due to organ specificity of MCs. In both dose groups, the highest concentrations of MCs (MC-RR + MC-LR) were found in blood at 1 h post-injection, 3757 ng/g DW (200 µg/kg dose group) and 270 ng/g DW (50 µg/kg dose group), respectively. MC-RR content in kidney was negatively correlated with that in blood in both dose groups, suggesting that blood was important in the transportation of MC-RR to kidney for excretion. After injection, liver to body weight ratio increased in crucian carp in both dose groups, especially in the 50 µg/kg dose group, which might be due to hydropic degeneration in liver. High depuration rate of MCs was observed in liver: after 48 h, 96.3% (200 µg/kg dose group) and 85.7% (50 µg/kg dose group) of the absorbed toxins were cleared. MCs accumulated in heart, kidney, gonad and brain at different levels, suggesting MCs might be toxic to these organs. Acute toxicity experiment in Japanese white rabbit: Japanese white rabbit were injected intraperitoneally with MC extract at two doses (50 and 12.5 μg MC-LReq./kg bw), and MC concentrations in various tissues were analyzed at 1, 3, 12, 24, and 48 h postinjection, using LC-MS. When rabbits were treated with 50 μg MC-LReq./kg bw via intraperitoneal injection, over half of the treated rabbits died at about 3 h post-injection, suggesting intense toxicity of MC to rabbit. MC contents in blood were very low in both dose groups, equal to only 0.02%-0.40% of the injected dose. In the 50 µg/kg dose group, the highest concentration of MCs was found in kidney, 62-158 ng/g DW, followed by liver, 54-82 ng/g DW. In the 12.5 µg/kg dose group, the highest concentration of MCs (MC-RR + MC-LR) was found in liver (15-65 ng/g DW) with a significant increase at 48 h compared with that at 1 h, suggesting that it was difficult for rabbit to clear up MCs in liver. Relative high MC content was observed in gastrointestinal tract and no decreace trend was showed in MC content in gallbladder and spleen, which might be attributed to the recirculation of adsorbed MC. MCs accumulated in lung, kidney and brain at different levels, suggesting that MCs might be toxic to these organs.
Document Type学位论文
Recommended Citation
GB/T 7714
雷和花. 微囊藻毒素在鲫鱼和日本长耳兔中组织分布的比较研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2008.
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