探討暴露聚苯乙烯塑膠微粒對小鼠大腦學習與記憶之分子機制與神經發炎的影響

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劉晏均(Yan-Jun Liu) 查詢紙本館藏

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生命科學系

論文名稱

探討暴露聚苯乙烯塑膠微粒對小鼠大腦學習與記憶之分子機制與神經發炎的影響
(The effects of polystyrene microplastics exposure on molecular mechanisms of learning and memory and neuroinflammation)

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★ 探討食入及吸入聚苯乙烯塑膠微粒對小鼠行為的影響

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摘要(中)

塑膠微粒(MPs)定義為直徑小於5 mm的塑膠顆粒，是一種新興環境污染物。因為塑膠微粒體積較小，它們能夠經由食物鏈進入生物體內，並積聚在周圍組織/器官中，隨後對器官造成損害。本實驗室先前研究發現，聚苯乙烯塑膠微粒(PS-MPs)會影響公鼠學習與記憶能力，然而，PS-MPs如何影響大腦學習及記憶的分子機制仍然未知。本篇研究結果顯示，小鼠口服2μm PS-MPs八周後，利用拉曼光譜分析，可觀察到2 μm PS-MPs在肝臟及大腦海馬迴腦區中累積，而海馬迴是學習與記憶的重要腦區，進一步針對八周PS-MPs公鼠海馬迴進行分子分析後，發現對突觸可塑性和記憶力至關重要的Activity-regulated cytoskeleton-associated protein (Arc)，其mRNA和蛋白質表達在海馬迴中顯著降低，但在PS-MPs母鼠中則沒有明顯差異。由於Arc蛋白會影響α-氨基-3-羥基-5-甲基-4-異噁唑丙酸受體(α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor, AMPAR)亞基之一的突觸蛋白GluA1的內吞作用，因此發現GluA1在八周PS-MPs公鼠的海馬迴中異常增加。此外，本篇研究也發現暴露PS-MPs的公鼠八周後會會增加海馬迴促炎因子TNF-α與IL-1β的mRNA表現量及小膠質細胞的活化。綜合上述結果顯示小鼠暴露PS-MPs會改變突觸相關蛋白的表達以及神經發炎，進而削弱海馬迴依賴性學習和記憶。

摘要(英)

Microplastics (MPs), defined as plastics particles less than 5 mm in diameter, are an emerging environmental pollutant in worldwide. Sine MPs are smaller in size, they can enter organisms via the food chain and accumulate in tissues/organs, subsequently causing damage to the organs. Previous study has confirmed that polystyrene micropartics (PS-MPs) can affect learning and memory ability in male mice. However, the molecular mechanism on how PS-MPs affecting learning and memory is still unknown. Using Raman spectroscopy analysis, 2 μm PS-MPs were found in the liver and the hippocampus of mice with oral administration of PS-MPs for eight weeks. Since the hippocampus is an important brain areafor learning and memory, Ifurther explored the molecular mechanism in the hippocampus. I found that the mRNA and protein expressions of activity-regulated cytoskeleton-associated protein (Arc), a key regulator for synaptic plasticity and learning and memory, was significantly decreased in the hippocampus, but there was no difference in PS-MPs female mice. The Arc protein can regulate α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) endocytosis. I found that GluA1, a subunit of AMPAR, was aberrantly increased in the hippocampus of PS-MPs male mice.Moreover, I also found that exposure to PS-MPs in male mice for eight weeks increased the mRNA expression of pro-inflammatory cytokines TNF-α and IL-1β in the hippocampus and induced microglia activation. Taken together, these results suggest that exposure to PS-MPs in mice alters synapse-associated protein expression and neuroinflammation, thereby impairing hippocampal-dependent learning and memory.

關鍵字(中)

★ 聚苯乙烯塑膠微粒
★ 海馬迴
★ 學習和記憶
★ 突觸可塑性分子Arc

關鍵字(英)

論文目次

中文摘要 i
英文摘要Abstract ii
聲明(Declaration) iii
誌謝 iv
目錄 v
圖目錄 viii
表目錄 ix
中英文對照表 x
一、緒論 1
1-1 塑膠微粒 1
1-2 塑膠微粒暴露的途徑 1
1-3 人類暴露塑膠微粒相關案例 3
1-4 暴露塑膠微粒對細胞分子層次的改變 4
1-5 聚苯乙烯塑膠微粒對周邊器官的影響-以無脊椎動物、魚類為例 5
1-6 聚苯乙烯塑膠微粒對周邊器官的影響-以齧齒類動物為例 6
1-7 聚苯乙烯塑膠微粒對中樞神經系統的影響-以無脊椎動物、魚類為例 8
1-8 聚苯乙烯塑膠微粒對中樞神經系統的影響-以齧齒類動物為例 8
1-9 聚乙烯塑膠微粒對中樞神經系統的影響-以齧齒類動物為例 9
1-10 研究動機與目的 9
二、研究方法 11
2-1 實驗動物 11
2-2 建立塑膠微粒動物模型 11
2-3 拉曼光譜之螢光PS-MPs組織萃取 11
2-4 西方墨點法 12
2-5 突觸神經體萃取 13
2-6 免疫螢光染色 14
2-7 RNA萃取 14
2-8 RNA定序(RNA -Sequencing) 14
2-9 定量聚合酶連鎖反應 15
2-10 統計分析 16
三、實驗結果 17
3-1 PS-MPs對於公和母鼠體重、運動能力及焦慮表現之影響 17
3-2 PS-MPs會累積至小鼠肝臟與大腦中 17
3-3 餵食PS-MPs之小鼠造成即刻早期基因(Immediate early gene, IEG)改變 18
3-4 攝入PS-MPs引起公鼠海馬迴神經突觸可塑性蛋白Arc表達改變 18
3-5 攝入PS-MPs引起公鼠海馬迴突觸蛋白的變化 19
3-6 攝入PS-MPs誘導公鼠海馬迴神經發炎 19
3-7 攝入PS-MPs誘導公鼠海馬迴小膠質細胞活化 20
四、討論 21
五、結論 25
參考文獻 38
附錄 44
附錄一、西方墨點法使用到之抗體 44
附錄二、免疫螢光染色使用到之抗體 45
附錄三、定量聚合酶連鎖反應使用到之Primer 46

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指導教授

黃佳瑜

審核日期

2022-7-26

推文