苯并（a）芘对人支气管上皮细胞BEAS-2B线粒体损伤的研究
发布时间：2018-08-01 16:41
【摘要】：[目的]研究致癌物质苯并(a)芘[Benzo(a)pyrene, B(a)P]对人支气管上皮细胞线粒体损伤的影响。 [方法]实验组分别用B(a)P浓度为50umol/1、1Oumol/1、1.Oumol/1的达尔伯克必需基本培养液(Dulbecco's Minimum Essential Medium,DMEM)对人支气管上皮细胞BEAS-2B进行培养,空白对照组用正常DMEM养液对人支气管上皮细胞BEAS-2B进行培养。培养24小时、48小时、72小时后,流式细胞仪测定细胞线粒体膜电位的变化情况；逆转录-聚合酶链反应(Reversed-transcription PCR, RT-PCR)检测线粒体细胞色素b(cytochrome b,Cyt-b)基因的表达情况。结果应用spss17.0统计软件进行统计学处理。 [结果]1.细胞线粒体膜电位的变化随培养时间延长,与空白对照组相比,实验组细胞线粒体膜电位水平逐渐下降,但50umol/1B(a)P、10umol/1B(a)P组细胞线粒体膜电位水平下降趋势更加明显(P0.01) 2.线粒体Cyt-b基因的表达细胞培养24h后,50umol/1B(a)P、10umol/1B(a)P组细胞Cyt-b mRNA相对表达量较空白对照组升高,50umol/l B(a)P组差异具有统计学意义(P0.05),10umol/1B(a)P、1.0umol/1B(a)P组表达较空白对照组无明显差异(P0.05)；细胞培养48h后,50umol/1B(a)P.10umol/1B(a)P组细胞Cyt-b mRNA相对表达量较空白对照组升高,差异具有显著统计学意义(P0.01),1.0umol/1B(a)P组表达较空白对照组无明显差异(P0.05)；细胞培养72h后,50umol/1B(a)P、10umol/1B(a)P组细胞Cyt-b mRNA相对表达量较空白对照组降低,50umol/1B(a)P组差异具有统计学意义(P0.05),1Oumol/1B(a)P、1.Oumol/1B(a)P组表达较空白对照组无明显差异(P0.05)。 [结论]B(a)P对人支气管上皮细胞BEAS-2B线粒体有损伤作用。B(a)P浓度越高,作用时间越长,损伤作用越强。
[Abstract]:[objective] to study the effect of benzo (a) pyrene [Benzo (a) pyrene, B (a) P] on mitochondrial damage in human bronchial epithelial cells. [methods] the human bronchial epithelial cells (BEAS-2B) were cultured in the experimental group with a concentration of 50 umoll / 1 Oumoll / 1 Oumoll / 1 Dalbek essential culture medium (Dulbecco's Minimum Essential Media DMEM), and the blank control group with the normal DMEM culture solution to culture the human bronchial epithelial cell BEAS-2B. After cultured for 24 hours and 48 hours for 72 hours, the changes of mitochondrial membrane potential and the expression of mitochondrial cytochrome b (cytochrome b cyt b gene were detected by flow cytometry and Reversed-transcription polymerase chain reaction (RT-PCR). Results spss17.0 statistical software was used for statistical processing. [result] 1. Compared with the blank control group, the level of mitochondrial membrane potential in the experimental group decreased gradually, but the decrease trend of the mitochondrial membrane potential in the 50umol/1B (a) 10 umol / 1B (a) P group was more obvious (P0.01) 2. Compared with the control group, the relative expression of Cyt-b mRNA in the mtDNA Cyt-b gene expression group was significantly higher than that in the control group (P0.05). There was no significant difference in the expression of Cyt-b mRNA between the 10 渭 mol / 1B (a) and 1.0 umolol / 1B (a) P group (P0.05). After 48 hours of cell culture, the relative expression of Cyt-b mRNA in 50 umol / 1B (a) P.10umol/1B (a) P group was higher than that in the blank control group, and the difference was statistically significant (P0.01). The expression of Cyt-b mRNA in 1.0 umol / 1B (a) P group was not significantly different from that in the blank control group (P0.05). After 72 hours of cell culture, there was no significant difference in the relative expression of Cyt-b mRNA between the control group and the control group (P0.05) compared with the control group (P0.05). The expression of Oumolol / 1B (a) / 1B (a) P in the Oumolr / 1B (a) P group was not significantly different from that in the blank control group (P0.05). [conclusion] B (a) P can damage BEAS-2B mitochondria of human bronchial epithelial cells. The higher the concentration of B (a) P is, the longer the time is and the stronger the damage is.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2158193
[Abstract]:[objective] to study the effect of benzo (a) pyrene [Benzo (a) pyrene, B (a) P] on mitochondrial damage in human bronchial epithelial cells. [methods] the human bronchial epithelial cells (BEAS-2B) were cultured in the experimental group with a concentration of 50 umoll / 1 Oumoll / 1 Oumoll / 1 Dalbek essential culture medium (Dulbecco's Minimum Essential Media DMEM), and the blank control group with the normal DMEM culture solution to culture the human bronchial epithelial cell BEAS-2B. After cultured for 24 hours and 48 hours for 72 hours, the changes of mitochondrial membrane potential and the expression of mitochondrial cytochrome b (cytochrome b cyt b gene were detected by flow cytometry and Reversed-transcription polymerase chain reaction (RT-PCR). Results spss17.0 statistical software was used for statistical processing. [result] 1. Compared with the blank control group, the level of mitochondrial membrane potential in the experimental group decreased gradually, but the decrease trend of the mitochondrial membrane potential in the 50umol/1B (a) 10 umol / 1B (a) P group was more obvious (P0.01) 2. Compared with the control group, the relative expression of Cyt-b mRNA in the mtDNA Cyt-b gene expression group was significantly higher than that in the control group (P0.05). There was no significant difference in the expression of Cyt-b mRNA between the 10 渭 mol / 1B (a) and 1.0 umolol / 1B (a) P group (P0.05). After 48 hours of cell culture, the relative expression of Cyt-b mRNA in 50 umol / 1B (a) P.10umol/1B (a) P group was higher than that in the blank control group, and the difference was statistically significant (P0.01). The expression of Cyt-b mRNA in 1.0 umol / 1B (a) P group was not significantly different from that in the blank control group (P0.05). After 72 hours of cell culture, there was no significant difference in the relative expression of Cyt-b mRNA between the control group and the control group (P0.05) compared with the control group (P0.05). The expression of Oumolol / 1B (a) / 1B (a) P in the Oumolr / 1B (a) P group was not significantly different from that in the blank control group (P0.05). [conclusion] B (a) P can damage BEAS-2B mitochondria of human bronchial epithelial cells. The higher the concentration of B (a) P is, the longer the time is and the stronger the damage is.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
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