纳米银对大鼠RG2胶质瘤细胞毒性作用的机制研究

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1、分类号：R651.1学校代码：10392学科专业代码：100210学号:2151003279硕士学位论文纳米银对大鼠RG2胶质瘤细胞毒性作用的机制研究TheMechanismofToxicityResearchoftheSilverNanoparticles(Ag-NPs)ontheRatGliomaCellsLineRG2学位类型：医学硕士所在学院：协和临床医学院申请人姓名：蔡然泽学科、专业：外科学（神经外科）导师：陈春美教授研究起止日期：2016年12月至2018年02月答辩委员会主席：林元相教授答辩日期：2018年05月18

2、日二○一八年五月1目录缩略词语„„„„„„„„„„„„„„„„„„„„„„„„3中文摘要„„„„„„„„„„„„„„„„„„„„„„„„„4英文摘要„„„„„„„„„„„„„„„„„„„„„„„„„6前言„„„„„„„„„„„„„„„„„„„„„„„„„„8材料与仪器„„„„„„„„„„„„„„„„„„„„„„„10实验方法„„„„„„„„„„„„„„„„„„„„„„„„13结果„„„„„„„„„„„„„„„„„„„„„„„„„„20讨论„„„„„„„„„„„„„„„„„„„„„„„„„„30结论„„„„„„„„„„„„„„„„„

3、„„„„„„„„„34参考文献„„„„„„„„„„„„„„„„„„„„„„„„35综述„„„„„„„„„„„„„„„„„„„„„„„„„„38致谢„„„„„„„„„„„„„„„„„„„„„„„„„„47研究生期间发表文章及获奖情况„„„„„„„„„„„„„„482缩略词语(Abbreviation)英文缩写英文全称中文全称Ag-NPsSilverNanpoparticles纳米银RG2RatGlioma2大鼠胶质瘤2型GBMGlioblastomamultiforme胶质母细胞瘤LGGsLowGradeGliomas低级别胶质瘤H

4、GGsHighGradeGliomas高级别胶质瘤ATCCAmericanTypeCultureCollection美国模式菌种收集中心DMEMDulbecco’smodifiedEagle’smedium培养基DMSODimethylsulphoxide二甲基亚砜CCK-8CellCountKit-8细胞计数试剂盒CCCytotoxicConcentrations细胞毒性浓度ROSReactiveOxygenSpecies活性氧簇TEMTransmissionElectronMicroscopy透射电子显微镜TBSTrietha

5、nolamineBufferedSaline三乙醇胺盐水缓冲液TBSTTriethanolamineBufferedSalineTween吐温三乙醇胺盐水缓冲液ODOpticalDensity吸光度PBSPhoshateBufferedSaline磷酸盐缓冲液FACSFluorescenceActivatedCellSorter免疫流式细胞检测FITCFluoresceinIsothiocyanate异硫氰酸荧光素PIPropidiumIodide碘化丙啶DCFDichlorofluorescein二氯荧光素DCFH-DADich

6、lorodihydrofluoresceinDiacetate二氯荧光黄双乙酸盐DNADeoxyribonucleicAcid脱氧核糖核酸hHour小时mgMilligram毫克mLMilliliter毫升μgMicroGram微克3纳米银对大鼠RG2胶质瘤细胞毒性作用的机制研究中文摘要目的胶质瘤是中枢神经系统最常见的原发性肿瘤，是一种侵袭性高、预后不良、术后复发率高的神经上皮源性肿瘤，临床上缺乏行之有效的治疗方法。胶质瘤恶性生长的基础是细胞增殖能力增强、细胞分化障碍和凋亡进程受阻，其中凋亡抑制因子bcl-2和凋亡活化因子casp

7、ase-3起着关键的作用。纳米银（Ag-NPs）作为一种新型抗肿瘤药物，具有稳定的物理化学特性、明显的肿瘤细胞组织穿透性与滞留效应、强大的表面修饰与负载性质、良好的生物相容性等特点。但是目前Ag-NPs是否对大鼠RG2神经胶质瘤细胞具有细胞毒性作用，及其作用的相关机制尚不明确。因此本研究建立体外Ag-NPs细胞毒性作用模型，以初步探讨Ag-NPs对胶质瘤细胞的相关毒性杀伤机制。方法本实验进行体外细胞研究，利用96孔和6孔培养板接种RG2细胞，加入不同浓度梯度的Ag-NPs（0、0.0125、0.025、0.05、0.1、0.2mg

8、/mL）连续培养，建立肿瘤细胞毒性杀伤模型。通过紫外-可见分光光度计和透射电镜（TEM）对Ag-NPs进行表征，观察其形貌、大小和分散度；通过CCK-8法检测Ag-NPs对RG2细胞增殖活性的影响；通过光学显微镜和TEM观察Ag-NPs对RG2细胞