摘要：黄曲霉是生活中重要的致病菌，能够引起人体发生全身系统性疾病，并导致食品发生腐败失去原有的营养价值。紫苏醛为紫苏中所提取的挥发油，具有一定的抗菌防腐作用。本实验通过检测不同浓度紫苏醛处理的黄曲霉胞内Ca2+浓度变化、活性氧的积累、线粒体膜电位、Cyt c的释放以及metacapase的活性，来探究Ca2+调控下紫苏醛对黄曲霉细胞凋亡的影响。实验结果表明：在一定浓度的紫苏醛（0。125 ~ 0。75 µl/ml）诱导下，黄曲霉胞内Ca2+浓度升高；并且在此基础上，细胞内活性氧发生积累、线粒体膜电位升高、Cyt c释放、metacapase活化。而且发现，经过较高浓度紫苏醛诱导的黄曲霉所得实验结果较其他浓度处理组更加明显。另外，我们通过TUNEL检测方法来观察细胞凋亡过程中细胞核DNA断裂情况，结果显示，紫苏醛诱导后的黄曲霉细胞中细胞核DNA发生断裂，且高浓度紫苏醛处理组结果效果更佳。从这些实验中可以看出，钙离子调控下紫苏醛对黄曲霉的细胞凋亡有着明显的调节作用。79901

毕业论文关键词：钙离子、紫苏醛、黄曲霉、细胞凋亡

Perillaldehyde triggers apoptosis in Aspergillus flavus via regulation of Ca2+

Abstract: Aspergillus flavus is an important pathogenic bacterium in our life, which can cause human systemic disease, leading to food corrupted and losing original nutritional value。 Perillaldehyde is volatile oil extracting from perilla, having some antibacterial preservative effect。 This experiment mainly test perillaldehyde at different concentrations of A。 flavus about Intracellular Ca2+ concentration, the accumulation of reactive oxygen species, mitochondrial membrane potential, Cyt c release and activity of metacapase, to explore effect of perillaldehyde on apoptosis of A。 flavus under calcium regulation。 Experimental results show that: in a certain concentration of perillaldehyde (0。125 ~ 0。75 μl/ml) induced A。 flavus intracellular Ca2+ concentration increasing。 And on this basis, intracellular reactive oxygen species accumulated, mitochondrial membrane potential increased, Cyt c released and metacapase was activated。 We found that after higher concentrations induced by perillaldehyde concentrations of A。 flavus from experimental results than other groups more visible。 In addition, we observe nucleus DNA fragmentation in apoptosis by TUNEL detection method。 This shows that nucleus DNA in A。 flavus was broken by perillaldehyde induction, and high concentration of perillaldehyde group’s result was better。 From these experiments can be seen, perillaldehyde induces apoptosis in A。 flavus via regulation of Ca2+。

Keywords: Calcium; Perillaldehyde; A。 flavus; Apoptosis。

目录

前言: 1

1 材料与方法 3

1。1 供试材料 3

1。1。1 原料 3

1。1。2 主要试剂 3

1。1。3 主要仪器 4

1。2 实验步骤 5

1。2。1 主要试剂与培养基的配制 5

1。2。2 配制培养基平板 5

1。2。3 黄曲霉活化与接种 6

1。2。4 孢子悬浮液的制备 6

1。2。5紫苏醛对黄曲霉细胞内钙离子浓度的影响 6

1。2。6 紫苏醛对黄曲霉细胞内活性氧积累的影响 6

1。2。7 紫苏醛对黄曲霉中线粒体膜电位的影响