水溶性聚醯胺乙二胺樹枝狀分子與碳六十複合體之合成及其作為抗氧化劑與基因載體之探討

Synthesis of Water-Soluble PAMAM Dendron-Bearing Fullerene conjugates as Antioxidant and Gene Carrier

洪澄祥(Cheng-Hsiang Hung)

指導教授 ： 朱智謙

中山醫學大學/健康管理學院/應用化學系碩士班/碩士(2012年)

https://doi.org/10.6834/CSMU.2012.00036

摘要

本研究成功地透過高效率的點擊化學反應合成出以poly (amido amine) (PAMAM) 樹枝狀高分子修飾的兩性C60/PAMAM複合體。此C60/PAMAM複合體具有高度的水溶性，其外圍PAMAM之胺基基團帶有多重正電荷，可與帶有負電荷之核酸結合以當作基因載體。透過乳癌細胞MCF-7與子宮頸癌細胞HeLa的細胞吞噬作用效率之探討，證明此C60/PAMAM複合體具有作為基因載體之能力。而在進一步的研究中，我們將plasmid DNA pEGFP-C1與C60/PAMAM複合體進行基因轉染實驗，發現具有穩定的綠色螢光蛋白GFP的表現。另外，由DPPH自由基清除測試、β-胡蘿蔔素漂白試驗法與Fenton氧化反應降解染料Rhodamine B的結果顯示，此C60/PAMAM複合體可作為有效的自由基清除劑。因此，本研究所合成出來之水溶性C60/PAMAM複合體可做為具有基因載體及抗氧化劑之雙功能新型態生醫材料。

關鍵字

富勒烯 ； 樹枝狀高分子 ； 點擊化學 ； 基因轉染 ； 抗氧化

並列摘要

In this research, we have successfully conjugated fullerene (C60) and poly (amido amine) (PAMAM) dendrons via efficient “click reaction” to yield amphiphilic fullerene-PAMAM-dendron bearing multiple positive charges. On the basis of strong binding between cationic C60 adducts and negatively charged nucleic acids, the amphiphilic C60/PAMAM dendron complex could serve as a non-viral vector toward in vitro gene transfection. Cellular uptake study shows that the complexes of C60/PAMAM dendrons surely deliver the pEGFP-C1 into HeLa and MCF-7 cell lines at low nitrogen-to-phosphorous ratio. Therefore, the MCF-7 cell lines could be successfully transfected with the pEGFP-C1, revealing a stable GFP expression. Moreover, DPPH radical scavenging assay, β-carotene bleaching assay, and the assessment of Rh-B decomposition via Fenton reaction indicate that the complexes could exhibit a remarkable ROS scavenging capability. Accordingly, the newly designed materials performed a dual function of antioxidant and gene carrier.

並列關鍵字

Fullerenes ； PAMAM dendrimers ； Click reaction ； ROS ； Gene delivery

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