白色念珠菌诱导ECV304细胞增殖的研究
发布时间：2019-06-03 02:25
【摘要】： 目的：通过提取白色念珠菌刺激成分与ECV304细胞相互作用,观察其对细胞增殖的影响,以解析白色念珠菌的致病性,以期揭示口腔念珠菌性白斑的发病机理。方法：体外培养人脐静脉内皮细胞(ECV304)模拟人口腔粘膜上皮细胞。培养白色念珠菌并提取白色念珠菌的上清液和灭活菌液,无菌验证后白色念珠菌刺激成分实验分组(上清液组、灭活菌液组、上清液和灭活菌液混合组)与ECV304细胞共培养。本课题具体从以下四个实验进行研究探索：1、通过倒置显微镜观察白色念珠菌上清液和灭活菌液对ECV304细胞密度变化的影响；2、通过MTT比色实验测定白色念珠菌刺激成分对ECV304细胞增殖的影响；3、通过细胞生长曲线测定白色念珠菌刺激成分对ECV304细胞增殖的影响；4、通过流式细胞术测定白色念珠菌刺激成分对ECV304细胞周期的影响。 结果：倒置显微镜观察发现4倍稀释的白色念珠菌上清液实验组细胞密度明显增高；而4倍稀释的白色念珠菌灭活菌液实验组细胞密度与对照组相比则无显著差异。MTT比色实验观察发现4倍稀释的白色念珠菌上清液实验组的细胞0D值与对照组相比差异最为显著。细胞生长曲线揭示在相同浓度、不同时间的培养条件下,随着时间的增长,不同组分对细胞促增殖影响基本呈正相关,以上清液组在作用48h时促增殖作用最为明显；流式细胞术研究发现上清液和灭活菌液分别作用细胞40h后,上清液组细胞S期、G2／M期所占百分比显著升高,反映细胞增殖活力指数PI增高,与对照组相比,有显著性差异(P＜O.05)。而灭活菌液组的PI值无显著性差异(乃O.05)。 结论：1.白色念珠菌的上清液和白色念珠菌的灭活菌液在一定程度上均可引起ECV30细胞增殖。实验表明,上清液作用明显大于菌体作用。 2.白色念珠菌的代谢产物可引起ECV304细胞周期的改变。 3.白色念珠菌的代谢产物可引起ECV304细胞的增殖。
[Abstract]:Aim: to investigate the pathogenicity of candida albicans by extracting the stimulating components of candida albicans and interacting with ECV304 cells in order to reveal the pathogenesis of oral candida leukoplakia. Methods: human umbilical vein endothelial cells (ECV304) were cultured in vitro to simulate human oral epithelial cells. The culture of candida albicans was cultured and the supernatant and inactivated liquid of candida albicans were extracted. After aseptic verification, the stimulating components of candida albicans were divided into three groups (culture fluid group, inactivated bacteria solution group, culture medium and inactivated liquid mixed group) and ECV304 cells were co-cultured. In this paper, the following four experiments were carried out: 1. The effects of culture medium and inactivated liquid of candida albicans on the density of ECV304 cells were observed by inverted microscope. 2. The effect of candida albicans stimulating components on the proliferation of ECV304 cells was determined by MTT colorimetric assay, and the effect of candida albicans stimulating components on the proliferation of ECV304 cells was determined by cell growth curve. 4. The effect of stimulating components of candida albicans on ECV304 cell cycle was determined by flow cytometry. Results: the cell density of the experimental group with 4 times dilution of candida albicans was significantly increased by inverted microscope. However, there was no significant difference in cell density between the experimental group and the control group. The cell density of the experimental group was not significantly different from that of the control group. MTT colorimetric assay showed that the cell 0D value of the experimental group was similar to that of the control group. The difference is the most significant. The cell growth curve showed that under the same concentration and different time culture conditions, with the increase of time, the effect of different components on the proliferation of cells was basically positively correlated, and the above liquid group had the most obvious effect on cell proliferation at 48 h. Flow cytometry showed that the percentage of S phase and G2 / M phase in the culture medium group was significantly higher than that in the control group, which reflected the increase of cell proliferation activity index (PI), which was significantly higher than that in the control group after 40 hours of treatment with the culture medium and the inactivated bacteria solution, respectively. the percentage of the cells in the S phase and G2 / M phase increased significantly. There was significant difference (P < 0.05). However, there was no significant difference in PI value in inactivated bacteria group (0.05). Conclusion: 1. To a certain extent, the culture fluid of candida albicans and the inactivated solution of candida albicans could induce the proliferation of ECV30 cells. The results showed that the effect of the culture medium was significantly greater than that of the bacteria. two銆，
本文编号：2491603
[Abstract]:Aim: to investigate the pathogenicity of candida albicans by extracting the stimulating components of candida albicans and interacting with ECV304 cells in order to reveal the pathogenesis of oral candida leukoplakia. Methods: human umbilical vein endothelial cells (ECV304) were cultured in vitro to simulate human oral epithelial cells. The culture of candida albicans was cultured and the supernatant and inactivated liquid of candida albicans were extracted. After aseptic verification, the stimulating components of candida albicans were divided into three groups (culture fluid group, inactivated bacteria solution group, culture medium and inactivated liquid mixed group) and ECV304 cells were co-cultured. In this paper, the following four experiments were carried out: 1. The effects of culture medium and inactivated liquid of candida albicans on the density of ECV304 cells were observed by inverted microscope. 2. The effect of candida albicans stimulating components on the proliferation of ECV304 cells was determined by MTT colorimetric assay, and the effect of candida albicans stimulating components on the proliferation of ECV304 cells was determined by cell growth curve. 4. The effect of stimulating components of candida albicans on ECV304 cell cycle was determined by flow cytometry. Results: the cell density of the experimental group with 4 times dilution of candida albicans was significantly increased by inverted microscope. However, there was no significant difference in cell density between the experimental group and the control group. The cell density of the experimental group was not significantly different from that of the control group. MTT colorimetric assay showed that the cell 0D value of the experimental group was similar to that of the control group. The difference is the most significant. The cell growth curve showed that under the same concentration and different time culture conditions, with the increase of time, the effect of different components on the proliferation of cells was basically positively correlated, and the above liquid group had the most obvious effect on cell proliferation at 48 h. Flow cytometry showed that the percentage of S phase and G2 / M phase in the culture medium group was significantly higher than that in the control group, which reflected the increase of cell proliferation activity index (PI), which was significantly higher than that in the control group after 40 hours of treatment with the culture medium and the inactivated bacteria solution, respectively. the percentage of the cells in the S phase and G2 / M phase increased significantly. There was significant difference (P < 0.05). However, there was no significant difference in PI value in inactivated bacteria group (0.05). Conclusion: 1. To a certain extent, the culture fluid of candida albicans and the inactivated solution of candida albicans could induce the proliferation of ECV30 cells. The results showed that the effect of the culture medium was significantly greater than that of the bacteria. two銆，
本文编号：2491603
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