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以牛血清白蛋白为硝化底物、过氧亚硝酸为硝化试剂,研究了褪黑素和芦丁对蛋白质硝基化的抑制作用.通过对硝基化反应各种影响因素的探讨,发现在37℃,pH 7.2,反应时间90 min以及过氧亚硝酸浓度为6.0×10-4mol/L时,硝基化反应进行最完全.实验分别在硝基化反应进行前、进行中以及反应后,在硝基化反应体系中引入不同浓度的抑制剂褪黑素和芦丁.结果显示,在硝基化反应发生前加入抑制剂,抑制硝基化反应的效果最为明显;当褪黑素和芦丁的终浓度为5.0×10-5mol/L时,抑制率分别达到80.7%和52.4%;若继续增加褪黑素,抑制率几乎不再变化,继续增加芦丁,抑制率小幅上升后开始呈下降趋势.总的来说,褪黑素抑制蛋白质硝基化的效果明显好于同浓度的芦丁.
Using bovine serum albumin as nitrification substrate and peroxynitrite as nitration reagent, the inhibitory effect of melatonin and rutin on the nitration of protein was studied.By investigating various influencing factors of nitration reaction, At 37 ℃, pH 7.2, reaction time 90 min and the concentration of peroxynitrite 6.0 × 10-4mol / L, the nitration reaction was the most complete.Every experiment was carried out before, during and after the nitration reaction, The different concentrations of inhibitors of melatonin and rutin were introduced into the nitration reaction system.The results showed that the addition of inhibitors before nitration reaction inhibited the nitration reaction most obviously.When melatonin and When the final concentration of Ding was 5.0 × 10-5mol / L, the inhibitory rates reached 80.7% and 52.4% respectively. If the melatonin was added, the inhibitory rate almost did not change any more, and the increase of rutin continued and the inhibitory rate increased slightly Down trend.In general, the effect of melatonin on inhibiting protein nitration is obviously better than that of rutin at the same concentration.