磺胺甲噁唑对绿藻的生长抑制作用及其潜在毒理学机制
New Insights into Growth Inhibition and Potential Toxicological Mechanisms of Sulfamethoxazole on Microalgae
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摘要: 磺胺甲噁唑(sulfamethoxazole, SMX)常用于动物和人类疾病治疗，导致其在多种环境介质中广泛检出。绿藻作为水生生态系统中的初级生产者，因其灵敏度较高而常用作毒性评估的模式生物。然而，目前关于SMX对绿藻的毒性研究普遍以传统毒性测试为主，缺乏对其生物大分子水平毒性效应的深入认识。本研究结合藻类生长抑制实验和傅里叶变换红外光谱(Fourier transform infrared spectroscopy, FTIR)分析深入探究SMX对绿藻生物大分子水平的毒性作用机制。藻类生长抑制实验结果表明，SMX对3株绿藻的EC50(置信度95%)依次为斜生栅藻(8.53 mg·L-1) < 四尾栅藻(14.30 mg·L-1) < 尖细栅藻(90.45 mg·L-1)。基于FTIR分析发现，SMX暴露引起绿藻生物大分子发生变化且存在明显的剂量效应关系。脂质水平上，脂质发生过氧化、改变脂肪酸烷基链长度，进而影响细胞膜的流动性和通透性。蛋白质水平上，酰胺Ⅰ和酰胺Ⅱ的变化揭示蛋白质二级结构发生变化。DNA水平上，DNA构型发生变化，由常见的B-DNA型转换为能抵御外界干扰A-DNA型。3株绿藻在生物大分子水平对SMX暴露的响应差异可能与其细胞壁结构组成差异有关。Abstract: Sulfamethoxazole (SMX) is widely used for human and veterinary medicine, leading to its frequent detection in various environmental matrices. Microalgae, as one of the major primary producers in the aquatic environment, have been commonly employed as indicators to evaluate the ecotoxicological effects of various environmental contaminants due to their high sensitivity. However, the toxicity mechanisms of SMX on microalgae have been usually evaluated by conventional toxicity tests, and little is known about its toxic mechanisms at the biomolecular level. In this study, the toxic mechanisms of SMX on microalgae at the biochemical level were investigated using the combination of the microalgal growth inhibition tests and Fourier transform infrared spectroscopy (FTIR) analysis. Based on microalgal growth inhibition tests, the EC50 (95% confidence intervals) of SMX to three tested microalgal species was observed with the following order: Scenedesmus obliquus (8.53 mg·L-1) < Scenedesmus quadricauda (14.30 mg·L-1) < Scenedesmus acuminatus (90.45 mg·L-1). Meanwhile, obvious concentration-related biomolecular alterations were induced by SMX exposure. Results suggested that SMX could cause the peroxidation of lipids and alterations in the length of lipid chains, leading to damage of the fluidity and permeability of cell membrane. Additionally, alterations in Amide Ⅰ and Amide Ⅱ indicated changes in the secondary structure of proteins. Furthermore, changes in DNA conformation were also induced by SMX, leading to the transition of B-DNA to A-DNA. Meanwhile, differences in FTIR spectral profiles caused by three tested microalgal species might be attributed to the differences in their cell walls composition and structure.
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