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目的研究次氯酸对人血清白蛋白(HSA)的氧化修饰影响及与高级氧化蛋白产物(AOPPs)之间的关系。方法有氧条件下在恒定浓度的HSA(60 mg／ml)内加入不同浓度次氯酸(0、1、5、10、20、30、40、50、60 mmol／L,最终浓度),观察氧化剂对HSA的修饰作用。凝胶排阻色谱法检验HSA氧化修饰结果,在线光谱扫描(190 nm～400 nm)分析修饰产物的光谱特性。结果次氯酸可氧化修饰人血清白蛋白,其修饰产物主要为二聚体HSA和六聚体HSA。发现次氯酸对HSA单体和HSA二聚体的氧化修饰为一级反应:对诱导生成的AOPPs为准一级反应:而对诱导生成的HSA六聚体则为二级修饰反应。同时发现白蛋白对AOPPs的主要贡献者是六聚体形式的HSA,光谱分析表明HSA聚集体的最大吸收峰发生红移,提示HSA聚集体是由于蛋白中的酪氨酸残基通过氧化交联方式而聚集形成的。结论HSA经次氯酸处理后主要发生了蛋白聚集。而对AOPPs的主要贡献者是六聚体形式的HSA。
Objective To study the effect of hypochlorous acid on the oxidative modification of human serum albumin (HSA) and its relationship with advanced oxidation protein products (AOPPs). Methods Hypoxia (0, 1, 5, 10, 20, 30, 40, 50 and 60 mmol / L, final concentration) was added into HSA (60 mg / Oxidants modify HSA. The results of the oxidative modification of HSA by gel exclusion chromatography were analyzed by online spectral scanning (190 nm ~ 400 nm). Results Hypochlorous acid oxidative modification of human serum albumin, its modified products are mainly dimer HSA and hexameric HSA. Hypochlorous acid was found to be a first order oxidative modification of HSA monomers and HSA dimers: a quasi-first order response to induced AOPPs and a second order modification reaction to induced HSA hexamers. At the same time, it was found that the major contributor to albumin to AOPPs was hexameric form of HSA. Spectral analysis showed a red shift in the maximum absorption peak of HSA aggregates, suggesting that HSA aggregates are due to oxidative cross-linking of tyrosine residues in proteins Way and gathered to form. Conclusion HSA mainly undergoes protein aggregation after hypochlorous acid treatment. The main contributor to AOPPs is the hexameric form of HSA.