寡核苷酸MTO1进入人牙周膜细胞的时相观察
发布时间：2018-10-18 12:21
【摘要】：实验目的： 探讨寡核苷酸MT01(一种根据人线粒体DNA序列设计的寡脱氧核苷酸)进入人牙周膜细胞的机制，为寡核苷酸MT01能够抑制牙槽骨吸收奠定实验基础。 实验方法： 体外培养并鉴定人牙周膜细胞，Cy5染色的MT01（工作浓度为1.0μg/ml）与人牙周膜细胞共培养15分钟，30分钟，1小时，3小时，6小时，12小时，24小时，36小时及48小时。激光共聚焦显微镜观察记录以上9个时间点MT01进入人牙周膜细胞的情况，对不同时间点人牙周膜细胞内MT01的荧光强度进行比对分析。采用SPSS17.0软件，配对样本t检验方法统计学分析两组间结果，P＜0.05有统计学差异，P＜0.01有显著统计学差异。 实验结果： 1.0μg/ml MT01进入人牙周膜细胞15分钟，30分钟，，1小时，3小时，6小时，12小时，24小时，36小时及48小时荧光强度分别为：1484±128，1771±206，2773±1364，1874±121，2437±293，3170±297，2442±273，2285±45，2119±374。其中12小时实验组荧光强度与15分钟、30分钟、6小时，48小时实验组荧光强度相比较均有显著差异（P＜0.01）。其余组间比较均无统计学差异。 实验结论： 1.寡核苷酸MT01无需载体,通过胞吞作用快速进入人牙周膜细胞胞质内，并且一定浓度的寡核苷酸MT01进入人牙周膜细胞呈时间依赖性：12小时达到峰值。 2.寡核苷酸MT01分布在人牙周膜细胞内两端，具有细胞极性。
[Abstract]:Objective: to investigate the mechanism of oligonucleotide MT01 (oligodeoxynucleotides) which is designed according to the sequence of human mitochondrial DNA (DNA) into human periodontal ligament cells, and to provide the experimental basis for the inhibition of alveolar bone resorption by oligonucleotide MT01. Methods: human periodontal ligament cells were cultured and identified in vitro. MT01 stained by Cy5 (working concentration was 1.0 渭 g/ml) was co-cultured with human periodontal ligament cells for 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours and 48 hours. Laser confocal microscopy observed the entry of MT01 into human periodontal ligament cells at the above nine time points and compared the fluorescence intensity of MT01 in human periodontal ligament cells at different time points. Using SPSS17.0 software and paired sample t-test method, the results of two groups were statistically different (P < 0. 05, P < 0. 01, P < 0. 01). The results showed that the fluorescence intensity of 1.0 渭 g/ml MT01 in human periodontal ligament cells for 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours and 48 hours was 1484 卤128,1771 卤2662773 卤1364U, 2437 卤2931737 卤2972442 卤2732442 卤273282285 卤451a 2119 卤3744h, respectively. The fluorescence intensity of the experimental group at 12 hours was significantly different from that of the experimental group at 15 minutes, 30 minutes, 6 hours and 48 hours (P < 0.01). There was no statistical difference between the other groups. Experimental conclusions: 1. Oligonucleotide MT01 was rapidly entered into the cytoplasm of human periodontal ligament cells by endocytosis without carrier, and a certain concentration of oligonucleotide MT01 entered into human periodontal ligament cells in a time-dependent manner: the peak value was reached at 12 hours. Oligonucleotide MT01 is distributed at both ends of human periodontal ligament cells and has cell polarity.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.4
本文编号：2279108
[Abstract]:Objective: to investigate the mechanism of oligonucleotide MT01 (oligodeoxynucleotides) which is designed according to the sequence of human mitochondrial DNA (DNA) into human periodontal ligament cells, and to provide the experimental basis for the inhibition of alveolar bone resorption by oligonucleotide MT01. Methods: human periodontal ligament cells were cultured and identified in vitro. MT01 stained by Cy5 (working concentration was 1.0 渭 g/ml) was co-cultured with human periodontal ligament cells for 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours and 48 hours. Laser confocal microscopy observed the entry of MT01 into human periodontal ligament cells at the above nine time points and compared the fluorescence intensity of MT01 in human periodontal ligament cells at different time points. Using SPSS17.0 software and paired sample t-test method, the results of two groups were statistically different (P < 0. 05, P < 0. 01, P < 0. 01). The results showed that the fluorescence intensity of 1.0 渭 g/ml MT01 in human periodontal ligament cells for 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 12 hours, 24 hours, 36 hours and 48 hours was 1484 卤128,1771 卤2662773 卤1364U, 2437 卤2931737 卤2972442 卤2732442 卤273282285 卤451a 2119 卤3744h, respectively. The fluorescence intensity of the experimental group at 12 hours was significantly different from that of the experimental group at 15 minutes, 30 minutes, 6 hours and 48 hours (P < 0.01). There was no statistical difference between the other groups. Experimental conclusions: 1. Oligonucleotide MT01 was rapidly entered into the cytoplasm of human periodontal ligament cells by endocytosis without carrier, and a certain concentration of oligonucleotide MT01 entered into human periodontal ligament cells in a time-dependent manner: the peak value was reached at 12 hours. Oligonucleotide MT01 is distributed at both ends of human periodontal ligament cells and has cell polarity.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.4
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