微米及奈米塑膠對瓶鼻海豚周邊血液單核細胞之影響

Effect of Microplastics and Nanoplastics on the Peripheral Blood Mononuclear Cells of Bottlenose Dolphins Tursiops truncatus

汪思宇(Szu-Yu Wang)

指導教授 ： 鄭謙仁

共同指導教授 ： 張惠雯(Hui-Wen Chang)

國立臺灣大學/獸醫專業學院/分子暨比較病理生物學研究所/碩士(2021年)

https://doi.org/10.6342/NTU202103170

摘要

現今塑膠垃圾污染嚴重，當塑膠垃圾流入海洋後，除了造成環境污染外，也會纏繞或因誤食進入海洋生物體內，造成海洋動物的傷害甚至死亡。塑膠性質穩定，且不易被大量分解，其易被降解成小片段塑膠或形成塑膠微粒，此時易被攝食進入海洋生物體內，造成許物理性、免疫性、代謝性和繁殖功能的損害；因此，評估塑膠微粒對海洋動物，特別是高階哺乳類的影響有其必要性。本篇論文利用遠雄海洋公園圈養瓶鼻海豚健康檢查所收集的血液，於實驗室分離周邊血液單核細胞(PBMC)，以兩種微米(Microplastics, MPs)及奈米(Nanoplastics, NPs)等級的塑膠微粒，在濃度10 μg/ml下共同培養。藉由染色、穿透式電顯及流式細胞儀和次世代定序分析評估細胞存活、氧化壓力誘導、細胞凋亡及細胞轉錄的改變。結果可觀察到在暴露不同濃度的塑膠微粒和奈米微粒並未造成PBMC的細胞毒性，即細胞存活率無明顯改變。再評估細胞凋亡方面，處理組隨同控制組因培養時間增加而有上升。此外，MPs可造成ROS 氧化壓力顯著增加。次世代定序mRNA 轉錄組分析結果顯示基因表現數量有發生改變，而且部分基因和免疫功能相關。在此，本研究顯示無修飾塑膠微粒和奈米微粒，在低劑量下仍會對瓶鼻海豚的PBMC免疫功能產生影響，因此，不應忽視其對鯨豚及高等哺乳類的衝擊。

關鍵字

瓶鼻海豚 ； 塑膠微粒 ； 奈米微粒 ； 細胞凋亡 ； 氧化壓力 ； 次世代定序

並列摘要

Nowadays, plastic wastes are stable and durable and are not easy to break down immediately. However, a slow degradation process could turn plastic wastes into small fragments or small particles which are easy to be ingested and may cause effects. Therefore, evaluation of the impact of plastic particles on the marine mammals is urgent. The aims of study were to investigate the effect of microplastics (MPs) and nanoplastics (NPs) on the peripheral blood mononuclear cell (PBMC) of bottlenose dolphins. PBMCs were co-cultured with 10μg/ml of MPs and NPs with diameter of 42 μm and 42 nm, respectively, for 24, 48, and 72h. The results showed MPs and NPs could be found intracellularly in PBMCs at each time point by light microscope and transmission electron microscope (TEM), respectively. Compared to the control group, no significant changes in viability, proliferation, and apoptosis were noticed in both MPs and NPs groups. However, significantly increased reaction oxygen species (ROS) production was only present in the MPs-treated group. In addition, the results of RNA sequencing showed significant changes in certain immune-related gene profiles the MPs and NPs treated group. The present study suggests, even in the low concentration of 10μg/ml, the impact of MPs and NPs on the PBMC is still present and should not be neglected

並列關鍵字

Cetacean ； microplastics ； nanoplastics ； apoptosis ； ROS ； NGS

參考文獻

Almeida, M., Martins, M. A., Soares, A. M. V., Cuesta, A., Oliveira, M. (2019). Polystyrene nanoplastics alter the cytotoxicity of human pharmaceuticals on marine fish cell lines. Environ Toxicol Pharmacol, 69, 57-65. doi:10.1016/j.etap.2019.03.019

Alshawafi, A., Analla, M., Alwashali, E., Ahechti, M., Aksissou, M. (2018). Impacts of Marine Waste, Ingestion of Microplastic in the Fish, Impact on Fishing Yield, M’diq, Morocco. International Journal of Marine Biology and Research, 3(2), 1-14. doi:10.15226/24754706/3/2/00125

An, R., Wang, X., Yang, L., Zhang, J., Wang, N., Xu, F., . . . Zhang, L. (2021). Polystyrene microplastics cause granulosa cells apoptosis and fibrosis in ovary through oxidative stress in rats. Toxicology, 449, 152665. doi:10.1016/j.tox.2020.152665

Andrady, A. L. (2011). Microplastics in the marine environment. Mar Pollut Bull, 62(8), 1596-1605. doi:10.1016/j.marpolbul.2011.05.030

Arthur, C., Baker, J. E., Bamford, H. A. (2009). Proceedings of the International Research Workshop on the Occurrence, Effects, and Fate of Microplastic Marine Debris, September 9-11, 2008, University of Washington Tacoma, Tacoma, WA, USA.