石墨烯-聚乙二醇-阿霉素复合物的制备及对淋巴瘤细胞的毒性研究

发布时间：2017-12-28 03:28

本文关键词：石墨烯-聚乙二醇-阿霉素复合物的制备及对淋巴瘤细胞的毒性研究　出处：《青岛大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的以淋巴瘤细胞系(Raji细胞)作为细胞模型,研究石墨烯-聚乙二醇-阿霉素复合物对淋巴瘤细胞的毒性,为纳米药物的应用提供理论依据。方法1、首先采用化学方法制备石墨烯-聚乙二醇(GO-PEG),然后将抗肿瘤药物阿霉素(DOX)负载到GO-PEG上获得石墨烯-聚乙二醇-阿霉素复合物(GO-PEG-DOX)。并通过透射电镜(TEM)、傅立叶分析变换红外光谱(FTIR)分别对GO及GO-PEG进行表征,通过紫外分光光度计(UV-Vis)检测GO-PEG负载DOX的载药程度。2、Raji细胞暴露于不同浓度的GO-PEG/GO-PEG-DOX(0-100μg/ml)中,分别在6h、12h、24h后,采用CCK-8法检测其对淋巴瘤细胞的毒性作用,并采用Annexin V-FITC法检测了GO-PEG(50μg/ml)/DOX(2μg/ml)/GO-PEG-DOX对淋巴瘤细胞的细胞凋亡的影响。结果成功制备获得石墨烯-聚乙二醇及石墨烯-聚乙二醇-阿霉素复合物,并证明其具有良好水溶性;实验结果显示石墨烯-聚乙二醇对淋巴瘤细胞的细胞毒性具有一定的时间和剂量依赖性,细胞毒性与石墨烯-聚乙二醇的浓度及接触时间呈正相关关系。GO-PEG组与空白对照组相比,P0.05,差异具有统计学意义。GO-PEG-DOX组与DOX组相比较,P=0.02,在统计学上具有显著性差异。但GO-PEG-DOX组与去除同浓度GO-PEG影响后与单纯阿霉素组相比较,p0.05,在统计学上无显著性差异。此实验结果表明GO-PEG不降低阿霉素对淋巴瘤细胞的细胞毒性;细胞凋亡检测结果显示阿霉素组/石墨烯-聚乙二醇-阿霉素组与正常对照组相比较,P均小于0.05,差异具有统计学意义。阿霉素组与石墨烯-聚乙二醇-阿霉素组比较显示P0.05,在统计学上无显著性差异。结论石墨烯-聚乙二醇对淋巴瘤细胞具有低毒性,石墨烯-聚乙二醇不降低阿霉素对淋巴瘤细胞的毒性。阿霉素可诱导淋巴瘤细胞发生细胞凋亡,但石墨烯-聚乙二醇不诱导恶性淋巴瘤细胞的凋亡,不影响阿霉素诱导细胞凋亡的作用。
[Abstract]:Objective to study the toxicity of graphene polyethylene glycol doxorubicin complex on lymphoma cells, using lymphoma cell line (Raji cell) as a cell model, and provide a theoretical basis for the application of nanomedicine. Method 1, first, we use graphene polyethylene glycol (GO-PEG) to prepare graphene polyethylene glycol (DOX). Then, the antitumor drug adriamycin (DOX) is loaded onto GO-PEG to obtain graphene polyethylene glycol doxorubicin complex (GO-PEG-DOX). The GO and GO-PEG were characterized by transmission electron microscopy (TEM), Fu Liye transform infrared spectroscopy (FTIR), and the loading degree of GO-PEG loaded DOX was detected by ultraviolet spectrophotometer (UV-Vis). 2, Raji cells were exposed to different concentrations of GO-PEG/GO-PEG-DOX (0-100 g/ml), 6h, 12h and 24h respectively after detecting the toxic effects on lymphoma cells by CCK-8 method, and using Annexin V-FITC method to detect the GO-PEG (50 g/ml) /DOX (2 g/ml) effects on cell apoptosis of lymphoma cell /GO-PEG-DOX. The result is successfully prepared by graphene and graphene - polyethylene glycol - polyethylene glycol doxorubicin complex, and prove that it has good water solubility; experimental results show that the graphene / PEG on lymphoma cell cytotoxicity with time and dose dependence, concentration and contact time was positively related with the cytotoxicity of graphene peg. The difference of P0.05 in group GO-PEG was statistically significant compared with that of blank control group. Group GO-PEG-DOX compared with group DOX, P=0.02, statistically significant difference. However, compared with the simple adriamycin group, there was no significant difference in P0.05 between the GO-PEG-DOX group and the simple adriamycin group after the removal of the same concentration of GO-PEG. The experimental results showed that GO-PEG did not reduce the cytotoxicity of adriamycin on lymphoma cells. The apoptosis assay showed that the P of doxorubicin group / graphene polyethylene glycol doxorubicin group was less than 0.05 compared with the normal control group, and the difference was statistically significant. There was no significant difference in P0.05 between adriamycin group and graphene PEG - adriamycin group. Conclusion graphene polyethylene glycol has low toxicity to lymphoma cells, and graphene polyethylene glycol does not reduce the toxicity of adriamycin to lymphoma cells. Doxorubicin can induce apoptosis in lymphoma cells, but graphene polyethylene glycol does not induce apoptosis in malignant lymphoma cells, and does not affect the apoptosis induced by adriamycin.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R733.1

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