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摘要：控制白色念珠菌引起的真菌感染是临床上较为棘手的一类难题，研究并开发新的有效的抗白色念珠菌药物成为了近年来抗真菌药物研发重点之一，而紫苏醛是一种成效显著、广谱、低毒的抗菌药物。本实验主要研究紫苏醛药液抗白色念珠菌的机制，观察不同浓度的紫苏醛药液对白色念珠菌的甾醇含量、细胞膜损伤，以简单研讨紫苏醛对真菌细胞膜的作用机理。用紫外可见分光光度测量麦角固醇含量，用流式细胞仪检测细胞膜损伤，结果显示紫苏醛可作用于细胞膜，抑制细胞膜中麦角固醇的合成，膜的通透性改变，损伤细胞膜。用流式细胞仪检测分析细胞周期和细胞凋亡，结果表明，紫苏醛能使细胞周期阻滞在G2/M期，诱导细胞发生凋亡。综上所述，不同浓度的紫苏醛均(0-0.4 µL/mL)对白色念珠菌的生长具有的抑制作用，抑制效果和浓度成正比，抗真菌机制主要归纳为抑制细胞膜中麦角固醇的合成导致细胞膜受损、阻滞细胞周期和诱导细胞凋亡。34461
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毕业论文关键词：紫苏醛；白色念珠菌；细胞周期；细胞凋亡
Influence ofperillaldehydeon cell cycle and apoptosis of Candida albicans
Abstract：Control the fungal infection caused byC.albicans is relatively difficult clinical problem. Consequently,research and development of new effective method of resistance of C.albicans has become one of the emphases on the antifungal drugs in recent yearsan,andperillaldehyde is a kind of effective, broad-spectrum, low toxicity of antibacterial drugs. Thisexperiment mainly study perillaldehyde solution mechanism of resistance of C.albicans,at different concentrations of perillaldehyde solution of C.albicans sterol content, cell membrane injury, in a simple discussion perillaldehyde on the mechanism of the fungal cell membrane. Ergosterol content is measured with a UV-spectrophotometer,usingflowcytometryinstrumentdetectioncell membrane damage, the results show that the perillaldehyde can be applied to the cell membrane, the synthesis of ergosterol in inhibition of cell membrane, change the permeability of membrane, damage the cell membrane. Using flow cytometry instrument testing analysis of cell cycle and cell apoptosis, test results indicate that, perillaldehyde can make the cell cycle arrest in GO/G1 phase, induction of cell apoptosis. In conclusion, different concentrations of perillaldehyde (0-0.4µL/mL) on the growth of C.albicans has inhibitory effect, inhibition effect and is directly proportional to the concentration, antifungal mechanism is mainly attributed to inhibit the synthesis of ergosterol in the cell membrane, destroy the integrity of the cell membrane, block the cell cycle and inducing apoptosis.
Key words：Perillaldehyde; C.albicans; Cell cycle; Apoptosis
目录
引言 1
1 材料与方法 3
1.1供试材料 3
1.1.1原料 3
1.1.2实验试剂与仪器 3
1.1.3 培养基 4
1.1.4 各种试剂溶液的配制 5
1.2 实验方法 6
1.2.1 白色念珠菌菌种活化 6
1.2.2 菌悬液的制备 6
1.2.3 微孔稀释法测定最低抑菌浓度(Minimum Inhibitory Concentration, MIC) 6
1.2.4紫苏醛对白色念珠菌甾醇含量的影响(Sterol Quantitation Method, SQM) 6
1.2.5紫苏醛对白色念珠菌细胞膜损伤的影响 7
1.2.6 紫苏醛对白色念珠菌细胞周期的影响 7
1.2.7 紫苏醛对白色念珠菌细胞凋亡的影响 8
2 结果与分析 9
2.1紫苏醛对白色念珠菌的MIC值 9
2.2紫苏醛对白色念珠菌甾醇含量分析 9
2.3紫苏醛对白色念珠菌细胞膜损伤测量 10