Improved method of plasma 8-Isoprostane measurement and association analyses with habitual drinking and smoking

• Published in: World journal of gastroenterology : WJG 2006-09, Vol.12 (36), p.5846-5852
• Main Authors: Kitano, Soichi, Hisatomi, Hisashi, Hibi, Nozomu, Kawano, Katsumi, Harada, Shoji
• Format: Article
• Language: English
• Subjects: Adult; Alcohol Drinking - blood; Biomarkers - blood; Clinical Research; Dinoprost - analogs & derivatives; Dinoprost - blood; Enzyme-Linked Immunosorbent Assay - methods; Female; Humans; Male; Sensitivity and Specificity; Smoking - blood; Solid Phase Extraction - methods; 等离子体; 过氧化反应
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v12.i36.5846

AIM： TO develop a simple and accurate method for quantifying 8-isoprostane in plasma by employing a combination of two-step solid-phase extraction of samples and a commercially available ELISA kit, and by this method to examine the effects of drinking and smoking habits against the levels of plasma 8-isoprostane in healthy Japanese volunteers.METHODS： Plasma 8-isoprostane was extracted with ODS gel suspension followed by NH2 Sep-Pak column. The 8-isoprostane fractions were assayed using a commercially available ELISA kit. We measured plasma 8-isoprostane levels in 157 healthy Japanese volunteers divided into three groups （64 non-habitual drinkers, 56 moderate drinkers and 37 habitual drinkers） according to their alcohol consumption per week. Genotypes of aldehyde dehydrogenase 2 （ALDH2） were also determined to investigate the plasma 8-isoprostane levels with reference to drinking habits. In addition, the plasma 8-isoprostane levels of 96 non-smokers and 61 smokers from the same subjects were compared.RESULTS： Our method fulfilled all the requirements for use in routine clinical assays with respect to sensitivity, intra- and inter-assay reproducibility, accuracy and dynamic assay range. Significant increases of plasma 8-isoprostane levels were observed in female habitual drinkers when compared with those of non-habitual drinkers （t = 5.494, P 〈 0.0001） as well as moderate drinkers （t = 3.542, P 〈 0.005）, and 8-isoprostane levels were also significantly different between ALDH2＊2/1 and ALDH2＊1/1 in the female habitual drinkers （t = 6.930, P 〈 0.0001）, suggesting that excessive drinking of alcohol may increase oxidization stress, especially in females. On the contrary, no significant difference of the plasma 8-isoprostane levels was observed between non-smokers and smokers.CONCLUSION： Our present method was proved to be a simple and accurate tool for measuring plasma 8-isoprostane. However, the clinical utility of plasma 8-isoprostane for drinking and smoking habits was limited since elevated 8-isoprostane levels were observed in female heavy drinkers, and no association was found between smokers and nonsmokers.