兰州大学机构库 >生命科学学院
癌性胸腔积液患者的氧化应激状态
Alternative TitleOxidative Stress in Cancer Patients With Malignant Pleural Effusion
刘晓菊
Thesis Advisor郑荣梁
2002-05-12
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword癌性胸腔积液 肿瘤伴随淋巴细胞 DNA 损伤 总抗氧化
Abstract本文对28 例经病理学证实而未经治疗的癌性胸腔积液患者,用不连续密度梯度离心法分离肿瘤伴随淋巴细胞(TAL),用单细胞凝胶电泳的方法检测外周血单个核细胞(PBMC)和TAL 的DNA 氧化损伤;采用菲罗啉比色法检测血浆及胸水上清的总抗氧化力(TAC);用硫代巴比妥酸比色法检测血浆脂质过氧化产物丙二醛(MDA);用邻苯二醛作为荧光标记的方法检测TAL 细胞内的还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)及其比值的变化。癌性胸腔积液患者血浆总抗氧化力(8.41±0.78 U/ml)明显低于健康对照组血浆(10.52±1.64 U/ml,p < 0.001)。患者胸水上清总抗氧化力(6.34±1.57 U/ml)显著低于自身血浆(p < 0.001)。总抗氧化力强度的顺序为:胸水上清< 患者血浆< 健康对照血浆,提示患者全身及肿瘤局部都处于氧化应激之中,肿瘤局部的氧化应激最为严重。癌性胸腔积液中TAL 的GSH 水平(0.97±0.20 mol/ 106cells)明显低于健康对照组PBMC(2.35±0.40 nmol/106cells,p < 0.001),GSH/GSSG 的比值(1.36±0.29)亦明显低于健康对照(3.33±0.58,p < 0.001),提示肿瘤患者TAL 也处于氧化应激中。 癌性胸腔积液患者血浆MDA(5.97±1.50 nmol/ml)明显高于健康对照组(3.84±0.52 nmol/ml,p < 0.001)。癌性胸腔积液患者血浆总抗氧化力与MDA 之间呈负相关(r = -0.516,p < 0.01)。再次说明癌性胸腔积液患者体内存在氧化应激,也再次说明肿瘤局部较全身明显。癌性胸腔积液患者PBMC 的DNA 损伤细胞%(16.8 ± 7.9)明显高于健康对照组PBMC(10.4 ± 4.9,p < 0.01);该组患者TAL 的DNA 损伤细胞%(41.9 ± 11.7)极显著高于自身PBMC 的DNA 损伤细胞%(p < 0.001)。细胞DNA 损伤程度依次为:患者TAL > 自身PBMC > 健康对照PBMC,再次提示癌性胸腔积液患者肿瘤局部氧化应激较全身明显。这组患者胸水上清总抗氧化力与TAL 的DNA 损伤细胞% 之间呈明显负相关(r =-0.538,p < 0.01),患者血浆总抗氧化力与患者PBMC 的DNA 损伤细胞%之间亦呈负相关(r = -0.426,p < 0.05);而患者血浆MDA 与PBMC 的DNA 损伤细胞% 之间呈正相关(r = 0.467,p < 0.05)。表明患者PBMC及TAL 两种细胞的DNA 损伤与氧化应激有很密切关系。分别用四种抗氧化剂N-乙酰半胱氨酸(NAC,5μmol/L)、褪黑素(10μmol/L)、槲皮素(5μmol/L)和亚硒酸钠(1.4μmol/L)处理TAL 4h后,DNA损伤细胞% 分别为23.7±10.7、27.1±9.9、28.6±12.3和47.0±10.6。与TAL对照组(40.4±11.5)比较,NAC和褪黑素可使TAL的DNA损伤细胞% 非常明显地降低(p < 0.001),槲皮素也可降低(p< 0.01),而亚硒酸钠则无显著影响(p > 0.05)。同时观察经上述处理后TAL细胞内GSH、GSSG和GSH/GSSG的变化,发现NAC、褪黑素、槲皮素都可使GSH和GSH/GSSG明显升高,其中褪黑素作用最显著(p < 0.001),GSSG基本不变。但亚硒酸钠可使GSSG增加(0.94±0.24,p < 0.05),GSH和GSH/GSSG无明显变化。NAC、褪黑素和槲皮素三种抗氧化剂对TAL的DNA损伤保护作用反证TAL的确处于氧化应激状态,而且其保护作用是通过提高细胞内的GSH和GSH/GSSG比值改变细胞内...
Other AbstractTAL were obtained by centrifugation on discontinuous density gradients from 28 patients with MPE, who comfired by cytology and were not under treatment in system and pleural cavity with anticancer agents during sample collection. DNA damages of peripheral blood mononuclear cells (PBMC) and of TAL are ssessed using single cell micro gel electrophoresis assay.Total antioxidant capacity was measured by o-phenanthroline colorimetric analysis.Malondialdehyde (MDA) was measured by thiobarbituric acid colorimetric analysis. GSH and GSSG assay were performed by fluorometrical analysis using o-phthaladehyde.Total antioxidant capacity of plasma is measured in 28 cancer patients with MPE and 33 healthy persons, and total antioxidant capacity of MPE supernatant is measured in these patients too. The total antioxidant capacity of plasma is remarkably lower in cancer patients (8.41±1.78 U/ml) than that in healthy persons (10.52±1.64 U/ml, p < 0.001). The total antioxidant capacity of MPE supernatant (6.34±1.57 U/ml) is significantly lower than that of plasma in patients (8.41±1.78 U/ml, p < 0.001). MDA (lipid peroxidation product) of plasma is significantly higher in cancerous patients with MPE (5.97±1.50 nmol/ml) than that in healthy persons (3.84±0.52 nmol/ml, p < 0.001). A negative relationship is observed between plasma total antioxidant capacity and MDA in cancerous patients with MPE (r = -0.516,p < 0.01).The number of DNA damaged cell percentage of PBMC is higher in cancer patients (16.8 ± 7.9) than that in healthy persons (10.4 ± 4.9, p < 0.01). Within cancer patients, the number of DNA damaged cell percentage of TAL (41.9 ± 11.7) is significantly higher than that of PBMC (16.8 ± 7.9, p < 0.001). A negative correlation is observed between total antioxidant capacity of MPE supernatant and the number of DNA damaged cell% of TAL in patients (r = -0.538, p < 0.01), and a negative corelation between total antioxidant capacity of plasma and the number of DNA damaged cell% of PBMC (r = -0.426, p < 0.05), as well as a positive corelation between MDA of plasma and number of DNA damaged cell % of PBMC (r = 0.467, p < 0.05) in these patients were found.After treated separately with N-acytelcysteine (NAC, 5 μ mol/L), melatonin(10 μ mol/L) and quercetin(5 μ mol/L), the number of DNA damaged cell% of TAL significantly declined to 23.7±10.7 (p < 0.001), 27.1 ±9.9 (p < 0.001), and 28.6±12.3 (p < 0.01), respectively. However, after treated with sodium selen...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221804
Collection生命科学学院
Recommended Citation
GB/T 7714
刘晓菊. 癌性胸腔积液患者的氧化应激状态[D]. 兰州. 兰州大学,2002.
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