【摘要】：[目的]研究致癌物質(zhì)苯并(a)芘[Benzo(a)pyrene, B(a)P]對(duì)人支氣管上皮細(xì)胞線粒體損傷的影響。 [方法]實(shí)驗(yàn)組分別用B(a)P濃度為50umol/1、1Oumol/1、1.Oumol/1的達(dá)爾伯克必需基本培養(yǎng)液(Dulbecco's Minimum Essential Medium,DMEM)對(duì)人支氣管上皮細(xì)胞BEAS-2B進(jìn)行培養(yǎng),空白對(duì)照組用正常DMEM養(yǎng)液對(duì)人支氣管上皮細(xì)胞BEAS-2B進(jìn)行培養(yǎng)。培養(yǎng)24小時(shí)、48小時(shí)、72小時(shí)后,流式細(xì)胞儀測(cè)定細(xì)胞線粒體膜電位的變化情況；逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(Reversed-transcription PCR, RT-PCR)檢測(cè)線粒體細(xì)胞色素b(cytochrome b,Cyt-b)基因的表達(dá)情況。結(jié)果應(yīng)用spss17.0統(tǒng)計(jì)軟件進(jìn)行統(tǒng)計(jì)學(xué)處理。 [結(jié)果]1.細(xì)胞線粒體膜電位的變化隨培養(yǎng)時(shí)間延長,與空白對(duì)照組相比,實(shí)驗(yàn)組細(xì)胞線粒體膜電位水平逐漸下降,但50umol/1B(a)P、10umol/1B(a)P組細(xì)胞線粒體膜電位水平下降趨勢(shì)更加明顯(P0.01) 2.線粒體Cyt-b基因的表達(dá)細(xì)胞培養(yǎng)24h后,50umol/1B(a)P、10umol/1B(a)P組細(xì)胞Cyt-b mRNA相對(duì)表達(dá)量較空白對(duì)照組升高,50umol/l B(a)P組差異具有統(tǒng)計(jì)學(xué)意義(P0.05),10umol/1B(a)P、1.0umol/1B(a)P組表達(dá)較空白對(duì)照組無明顯差異(P0.05)；細(xì)胞培養(yǎng)48h后,50umol/1B(a)P.10umol/1B(a)P組細(xì)胞Cyt-b mRNA相對(duì)表達(dá)量較空白對(duì)照組升高,差異具有顯著統(tǒng)計(jì)學(xué)意義(P0.01),1.0umol/1B(a)P組表達(dá)較空白對(duì)照組無明顯差異(P0.05)；細(xì)胞培養(yǎng)72h后,50umol/1B(a)P、10umol/1B(a)P組細(xì)胞Cyt-b mRNA相對(duì)表達(dá)量較空白對(duì)照組降低,50umol/1B(a)P組差異具有統(tǒng)計(jì)學(xué)意義(P0.05),1Oumol/1B(a)P、1.Oumol/1B(a)P組表達(dá)較空白對(duì)照組無明顯差異(P0.05)。 [結(jié)論]B(a)P對(duì)人支氣管上皮細(xì)胞BEAS-2B線粒體有損傷作用。B(a)P濃度越高,作用時(shí)間越長,損傷作用越強(qiáng)。
[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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

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