本文選題：香煙煙霧提取物CSE　切入點：彗星實驗　出處：《武漢科技大學》2012年碩士論文

【摘要】：目的 香煙燃燒產物中含有多種致突變物和致癌物，可以對人體產生嚴重的遺傳損傷，本實驗通過提取香煙燃燒產物制作香煙煙霧提取物，并用它刺激肺泡上皮細胞，觀察香煙對肺泡上皮細胞的DNA損傷，初步探討香煙導致肺泡上皮細胞遺傳損傷。 方法 培養(yǎng)人肺泡上皮細胞A549，以二甲基亞砜(DMSO)作為溶劑制取香煙煙氣提取物(cigarette smoke extract,CSE)，將不同濃度的CSE作用于A549細胞，用MTT法測定細胞活性，檢測DNA-蛋白質交聯(lián)率（DNA-protein crosslinks,DPC)及彗星實驗觀察DNA損傷，同時檢測超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶（GSH-PX）、丙二醛（MDA）觀察CSE對肺泡細胞的氧化損傷作用。 結果 1.10%、20%、50%CSE作用組與對照組相比細胞活性無明顯差異，，（P㧐0.05）活性無明顯影響，而100%CSE組的細胞活性則明顯下降且差異有統(tǒng)計學意義（P㩳0.05）； 2.CSE各濃度組與空白對照組及溶劑組相比，尾部DNA含量，尾長，尾距，Olive尾距均明顯增高，差異有統(tǒng)計學意義（P㩳0.05），20%CSE及以上濃度組均表現(xiàn)DPC增高，兩指標均有劑量效應關系。 3.20%CSE及以上劑量組的SOD、GSH-PX與對照組相比明顯下降， MDA則上升，（P㩳0.05），有一定的劑量效應關系。 4.DPC與抗氧化指標之間存在相關性（P㩳0.05）。 結論 1.CSE可導致A549細胞內氧化損傷增加。 2. CSE可導致A549細胞DNA損傷。 3. CSE可導致A549細胞內氧化損傷指標和其導致的DPC率存在相關性。
[Abstract]:Purpose. Cigarette burning products contain many mutagens and carcinogens, which can cause serious genetic damage to human body. In this experiment, cigarette smoke extracts were extracted from cigarette burning products and stimulated alveolar epithelial cells. To observe the DNA damage of alveolar epithelial cells induced by cigarette, and to explore the genetic damage of alveolar epithelial cells induced by cigarette. Method. Human alveolar epithelial cells (A549) were cultured with dimethyl sulfoxide (DMSO) as solvent to prepare cigarette smoke extract smoke extract CSE. Different concentrations of CSE were treated on A549 cells. The cell activity was determined by MTT assay. DNA-protein crosslinking rate (DNA-protein crosslinking DPCs) and comet assay were used to observe the damage of DNA, superoxide dismutase (SOD), glutathione peroxidase (GSH-PXX) and malondialdehyde (MDA) to observe the oxidative damage of CSE to alveolar cells. Results. There was no significant difference in cell activity between CSE group and control group. The cell activity of 100%CSE group was significantly decreased and the difference was statistically significant. 0.05; Compared with the control group and solvent group, the DNA content, tail length, tail distance and Olive tail distance of 2.CSE group were significantly higher than those of control group and solvent group, and the difference was statistically significant. The concentration of DPC was increased in the group of 0. 05 and 20% CSE, both of which had dose-effect relationship. Compared with the control group, the GSH-PX of 3.20%CSE and above dose group decreased significantly, while MDA increased. There is a dose-effect relationship between the two groups. There is a correlation between 4.DPC and antioxidant index. 0.05. Conclusion. 1.CSE induced increased oxidative damage in A549 cells. 2. CSE induced DNA damage in A549 cells. 3. There was a correlation between the index of oxidative damage and the rate of DPC induced by CSE in A549 cells.
【學位授予單位】：武漢科技大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R114

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