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摘要：植物挥发油是从植物的根、茎、叶或果实中提取出来芳香物质，它用途广泛，含有多种生物活性物质，许多植物挥发油可以用于生物制药。有研究表明，一些植物挥发油具有抑制肿瘤细胞生长，诱导细胞凋亡的作用。紫苏在我国种植广泛，历史悠久，在食品和医药领域有着重要的价值。本次实验以来自我国十个不同产地的紫苏油为研究对象，探究其是否具有抗肿瘤活性及其可能的机制。35826
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实验分为MTT实验、台盼蓝染色、Western Blot几个部分。MTT实验、台盼蓝染色用于检测紫苏油对A549细胞活性的影响。实验结果表明，各个产地的紫苏油处理细胞后，细胞的死亡率显著增加。不同产地紫苏油的处理效果有所差别，其中贵州、甘肃紫苏油的处理效果最明显。通过计算各产地紫苏油的IC50，发现贵州紫苏油的IC50值最小，仅为0.039 μl/ml。为探究紫苏油的抗肿瘤机理，进一步用Western Blot方法检测紫苏油对凋亡相关蛋白caspase-8、caspase-3、Bcl-2、Bcl-XL、PARP、HSP70、P53等的影响。研究发现HSP70上调，caspase-8、caspase-3酶原被切割，Bcl-2、Bcl-XL下调，P53含量增加，PARP被切割后失活。以上实验结果表明，紫苏油具有潜在的抗肿瘤活性，且可以促进肿瘤细胞A549的凋亡。在紫苏油的作用下，caspase蛋白酶逐渐被切割活化，线粒体外膜通透性增加，启动外源与内源性的caspase依赖性凋亡途径，使A549细胞凋亡。
毕业论文关键词：紫苏油；A549细胞；细胞凋亡；抗肿瘤
Studies on antitumor activity of Perilla frutescens essential oil
Abstract: Plant essential oils are aromatic substances extracted from plant roots, stems, leaves and fruits. These essential oils used widely and have a variety of biological activity, especially used for drugs production. Recently, it has been confirmed that some plant essential oils can inhibit tumor cell growth and induce cell apoptosis. Perilla frutescens were widely planted in our country with a long history. It has high application value in the field of food and medicine. This experiment use P. frutescens essential oils from ten different origins as the experimental subject, to explore whether they have antitumor activity and possible mechanism.
Experiment is pided into MTT assay, trypan blue dye exclusion assay and western blot. MTT assay and trypan blue dye exclusion assay was used to detect the activity of A549 cells incubated with P. frutescens essential oils. The results show that the mortality rate of cell treated with P. frutescens essential oils increased significantly. Treatment effect of essential oils differs dramatically. Effects of Guizhou and Gansu P. frutescens essential oils are obvious. By calculating IC50 of each essential oil show that IC50 value of Guizhou P. frutescens essential oil is minimum, only 0.039μl/ml. To explore the anti-tumor mechanism of essential oils, we perform western blot to detect the change of protein concentration, such as caspase-8, caspase-3, Bcl-2, Bcl-XL, PARP, HSP70 and P53. HSP70 is increased, caspase-8, caspase-3 zymogens was cut, the concentrations of Bcl-2, Bcl – XL are decreased, P53 is increased, and PARP was cleaved and deactivated. These results showed that essential oils have potential antitumor activity by promoting the apoptosis of tumor cells. Apoptosis of A549 cells was induced by P. frutescens essential oils through the activation of the exogenous and endogenous caspase-depended of apoptotic pathway, which was induced by the activation of the cleaved caspase protease and the increase of the mitochondrial membrane permeability.
Key words: Perilla frutescens essential oils; A549 cell; apoptosis; antitumor
目录
引言 1
1 材料与方法 2
1.1供试材料 2
1.2细胞 2
1.3试剂 2
1.4细胞培养 2
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