吡唑醚菌酯对斑马鱼早期生命阶段的亚致死效应研究
Sublethal Effects of Pyraclostrobin on Zebrafish during Early Life Stages
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摘要: 为明确吡唑醚菌酯对斑马鱼早期生命阶段的亚致死效应和慢性毒性，研究了不同浓度吡唑醚菌酯(0.328~32.0 μg·L-1)长期暴露斑马鱼胚胎30 d后，对斑马鱼生长发育，以及氧化胁迫、能量代谢和肝脏能量代谢酶活等生化指标的影响。研究发现，吡唑醚菌酯暴露30 d后，5.12 μg·L-1吡唑醚菌酯能够显著降低斑马鱼仔鱼的体长，2.05、5.12和12.8 μg·L-1吡唑醚菌酯长期暴露后可以显著增加仔鱼的体质量，0.819、5.12和12.8 μg·L-1吡唑醚菌酯可以引起仔鱼肝脏不同程度的空泡化。此外，为了进一步明确吡唑醚菌酯对斑马鱼胚胎发育过程中的毒性效应，分别于暴露2 d、6 d、10 d、16 d和30 d测定斑马鱼胚胎/仔鱼体内的丙酮酸、葡萄糖、胆固醇、甘油三酯、游离脂肪酸含量，以及与氧化胁迫和肝脏代谢相关的酶活。在30 d暴露期间，吡唑醚菌酯对斑马鱼过氧化氢酶(CAT)活性总体呈现先降低再升高的趋势，对过氧化物歧化酶(SOD)和游离脂肪酸也有不同程度的影响，吡唑醚菌酯处理对斑马鱼体内丙酮酸含量总体呈现先降低后升高的趋势，对甘油三酯含量总体呈现降低的趋势，而吡唑醚菌酯对不同生长阶段斑马鱼体内的葡萄糖和总胆固醇合成有一定的诱导作用，暴露6 d和30 d时所有吡唑醚菌酯处理组鱼体内葡萄糖含量显著增加。此外，吡唑醚菌酯对斑马鱼早期生命阶段的碱性磷酸酶(AKP)活性有一定的诱导作用，对谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性也有不同程度的影响。研究表明，吡唑醚菌酯可能通过诱导氧化应激、干扰葡萄糖、总胆固醇等糖类和脂类的合成，影响斑马鱼体内抗氧化系统和能量代谢，引起肝脏能量代谢相关酶的活性改变进一步干扰能量代谢，进而导致斑马鱼早期生命阶段的生长和肝脏发育异常。Abstract: To assess the sublethal effects and chronic toxicity of pyraclostrobin on zebrafish early life stage, the impact on the growth, oxidative stress, energy metabolism and enzymes involved in liver energy metabolism were determined after zebrafish embryos exposed to various concentrations (0.328~32.0 μg·L-1) of pyraclostrobin for 30 d. Result showed 5.12 μg·L-1 pyraclostrobin reduced the body length of larvae, 2.05, 5.12, and 12.8 μg·L-1 pyraclostrobin increased the body weight after 30 d exposure, while 0.819, 5.12 and 12.8 μg·L-1 pyraclostrobin caused different degrees of vacuolation in larvae liver. To further clarify the toxic effects of pyraclostrobin during zebrafish embryonic development stages, the contents of pyruvate, glucose, cholesterol, triglycerides and free fatty acids, and the enzymes related to oxidative stress and liver energy metabolism were determined at 2 d, 6 d, 10 d, 16 d, and 30 d. During the exposure, it was found the activities of catalase (CAT) first decreased and then increased, exposure to pyraclostrobin also affected the peroxide dismutase (SOD) activity and free fatty acids level during zebrafish different early life stages, the content of pyruvate decreased first then increased, the triglyceride levels decreased overall during 30 d exposure, while pyraclostrobin induced glucose and total cholesterol levels during zebrafish early different life stages, significantly increased the glucose levels at 6 d and 30 d. In addition, pyraclostrobin induced the alkaline phosphatase (AKP) activity during the early life stages of zebrafish, also affected the alanine aminotransferase (GPT) and glutamic-oxaloacetic transaminase (GOT) activities during 30 d exposure. The results suggested pyraclostrobin might induce oxidative stress, interfered with the synthesis of glucose and total cholesterol, affected the antioxidant system and energy metabolism in zebrafish, then altered the enzyme activity involved in liver energy metabolism further disturbed the energy metabolism, which caused negative effects on the growth and liver development during the early life stages of zebrafish.
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Key words:
- pyraclostrobin /
- zebrafish /
- chronic toxicity /
- early life stage
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