低强度激光对细胞端粒DNA长度影响的实验研究

发布时间：2018-01-25 13:14

本文关键词： 氦氖激光人胚肺二倍体成纤维细胞端粒长度实时荧光定量PCR　出处：《广西医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：本实验将体外培养的人胚肺二倍体成纤维细胞(Human fetal lung diploid fibroblasts2BS)作为细胞衰老模型,开展低强度激光对成纤维细胞端粒DNA长度影响的实验研究。采用不同辐照时间、不同辐照功率的低强度激光辐照实验组细胞即年轻人胚肺二倍体成纤维细胞,用四甲基偶氮唑盐(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltrazolium bromide MTT)比色法检测激光照射处理对细胞生长和增值的影响；采用相对定量实时荧光聚合酶链式反应(Quantitative Real-time Polymerase Chain Reaction q-PCR)方法检测细胞端粒DNA相对长度。结果如下： (1)MTT比色法检测细胞生长与增值的结果显示,激光照射后,实验组(即激光照射组)(功率2-5mmw)细胞生长趋势显著优于对照组(非激光照射组)(P0.05)；提示激光照射对细胞生长不但没有对细胞造成负影响,而是促进细胞生长。 (2) q-PCR检测细胞端粒DNA长度显示：对照组中,老年2BS细胞端粒DNA长度较年轻2BS细胞端粒DNA短(P0.05),进一步验证细胞端粒DNA长度随着代龄的增加而缩短,从而也说明了端粒与细胞衰老息息相关。实验组中,老年细胞(2BS细胞在年轻时经激光照射后传代至老年时)的端粒长度较对照组的老年细胞端粒长度长(P0.05)。提示,经适当激光照射后,细胞端粒DNA因衰老而变短的趋势得到减缓。本研究从基因水平上为探讨低强度激光延缓细胞衰老的激光生物效应提供实验依据。本实验首次将激光照射应用于人类细胞衰老模型,同时运用最简便、快捷、专一、灵敏、可重复性强的检测细胞端粒DNA长度的方法——实时荧光定量PCR方法来检测细胞端粒DNA长度,提供低强度激光延缓细胞衰老的生物物理学实验依据和生物分子学理论基础,在细胞基因水平上探讨低强度激光的抗衰老机理。同时,通过实验筛选有生物意义的激光参数,为低强度激光在延长细胞寿命、预防及降低与年龄相关性疾病的发生等方面的应用提供新途径和新技术。
[Abstract]:In this study, human fetal lung diploid fibroblast 2BS (BSs) of human fetal lung diploid fibroblasts cultured in vitro was used as the model of cell senescence. The effects of low intensity laser on the length of telomere DNA of fibroblasts were studied. The experimental group cells were irradiated with low intensity laser with different irradiation power, I. e., diploid fibroblasts of young embryo lung, treated with tetramethylazolium trimethylazolium. 5-Dimethylthiazol-2-yll-2. The effects of laser irradiation on cell growth and proliferation were detected by 5-diphenyltrazolium bromide MTT colorimetry. The relative quantitative real-time fluorescence polymerase chain reaction (. Quantitative Real-time Polymerase Chain Reaction q-PCR). Methods the relative length of telomere DNA was measured. The results were as follows: Cell growth and proliferation were detected by MTT colorimetry. The growth trend of the cells in the experimental group (i.e. the laser irradiation group) (power 2-5mmw) was significantly better than that in the control group (non-laser irradiation group). The results suggest that laser irradiation not only has no negative effect on cell growth, but also promotes cell growth. The length of telomere DNA was detected by q-PCR: in the control group, the telomere DNA length of the aged 2BS cells was shorter than that of the younger 2BS cells (P 0.05). Further verify that the length of telomere DNA is shortened with the increase of age, which also shows that telomere is closely related to cell senescence. The telomere length of the geriatric cells was longer than that of the control group (P 0.05). It suggested that the telomere length of the geriatric cells after laser irradiation was higher than that of the control group (P 0.05). The results suggested that the telomere length of the geriatric cells was higher than that of the control group. The trend of telomere DNA shortening due to aging has been slowed down. This study provides an experimental basis for the study of laser biological effects of low-intensity laser on delaying cell senescence at the gene level. In this experiment, laser irradiation was applied to human aging model for the first time, and it was the most convenient, rapid, specific and sensitive. A reproducible method for detecting the length of telomere DNA in cells-real-time fluorescent quantitative PCR method is used to detect the length of telomere DNA. To provide the experimental basis of biophysics and biomolecular theory of low-intensity laser to delay cell senescence, and to explore the anti-aging mechanism of low-intensity laser on the level of cell gene. In order to provide new approaches and techniques for the application of low intensity laser in prolonging cell life, preventing and reducing the occurrence of age-related diseases and so on.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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