SSAO介导的氧化脱氨反应对3T3-L1脂肪细胞的氧化应激作用  

Effect of deamination reaction mediated by semicarbazide-sensitive amine oxidase on 3T3-L1 adipocytes

作　　者：张琼丽[1] 罗红军[1] 李慧[1] 林哲绚[1] 罗文鸿[1] 

机构地区：[1]汕头大学医学院生物分析实验室,广东汕头515041

出　　处：《中国病理生理杂志》2015年第3期468-474,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81170315);广东省自然科学基金团队资助项目(No.9351507102000-001)

摘　　要：目的：观察氨基脲敏感型胺氧化酶（SSAO）催化其底物甲胺（MA）和苯甲胺（BZA）的氧化脱氨反应对成熟3T3-L1脂肪细胞的氧化应激作用。方法：3T3-L1前脂肪细胞诱导分化为成熟脂肪细胞;高效液相色谱法检测不同分化天数下细胞SSAO活性的变化;MTT法检测不同浓度的MA或BZA对细胞活力的影响;荧光探针方法检测细胞在药物作用下产生的活性氧;以0.5 mmol/L MA或BZA处理成熟脂肪细胞或未经诱导分化的前脂肪细胞4 h,观察细胞产生的甲醛（FA）、苯甲醛（BZ）、脂质过氧化指标丙二醛（MDA）、总超氧化物歧化酶（T-SOD）和谷胱甘肽（GSH）的变化。结果：SSAO活性随细胞分化天数的增加而增加,并于第8天达到高峰;不同浓度MA或BZA作用细胞4 h对细胞活力无显著影响（P〉0.05）;0.5 mmol/L MA或BZA孵育后,细胞产生的活性氧增高,约为阴性对照组的3~4倍,差异有统计学意义（P〈0.05）;在成熟脂肪细胞中,MDA的含量增加,而T-SOD和GSH的活性和含量减少,与阴性对照组相比,差别有统计学意义（P〈0.05）;MA和BZA作用未经诱导分化的前脂肪细胞,其MDA和SOD和GSH的变化不明显,差别无统计学意义（P〉0.05）。结论：SSAO可能通过其介导的氧化脱氨反应引起3T3-L1成熟脂肪细胞产生氧化应激。AIM：To observe the effect of the metabolites generated from oxidative deamination of methylamine （ MA） or benzylamine （ BZA ） catalyzed by semicarbazide-sensitive amine oxidase （ SSAO ） on 3T3-L1 adipocytes. METHODS：3T3-L1 preadipocytes were induced to differentiation.SSAO activity was determined by high performance liquid chromatography （ HPLC） at different differentiation time points.MTT assay was applied to detect cell vitality after exposure to different concentrations of MA or BZA.Fluorescence probe DCFH-DA was used to determine the production of reactive oxygen species after incubation of 3T3-L1 adipocytes with MA or BZA.After exposure to 0.5 mmol/L MA or BZA for 4 h, malondialdehyde （ MDA） , total superoxide dismutase （ T-SOD） and glutathione （ GSH） in the adipocytes or prea-dipocytes were measured.RESULTS：SSAO activity increased with the increase in the differentiation days, and reached a maximum at the 8th day.Incubation of the cells with different concentrations of MA or BZA for 4 h did not significantly de-creased the cell vitality （P〉0.05）.After exposure to 0.5 mmol/L MA or BZA, the reactive oxygen species in adipocytes significantly increased, and were about 3 to 4 times as compared with control group （P〈0.05）.After treatment with 0.5 mmol/L MA or BZA for 4 h, MDA content significantly increased, while the activity of SOD and the expression of GSH de-creased in mature adipocytes compared with control group （P〈0.05）.However, MDA, T-SOD and GSH did not change significantly after treatment with equal molar of MA or BZA in the preadipocytes （ P〉0.05 ） .CONCLUSION： MA or BZA induces oxidative stress in the mature adipocytes, which might result from the deamination products catalyzed by SSAO.

关 键 词：氨基脲敏感型胺氧化酶 脂肪细胞 甲胺 苯甲胺 氧化应激 

分 类 号：R363[医药卫生—病理学]