羟基红花黄色素A对脑组织蛋白质羰基化影响的体外研究

摘要

目的 研究羟基红花黄色素A(HSYA)对体外ONOO-途径和亚铁血红素/亚硝酸钠/过氧化氢(heme/NaNO2/H2O2)途径引起脑组织蛋白质羰基化的影响.方法 分别模拟体内ONOO-、heme/NaNO2/H2O2羰基化途径,以脑组织蛋白为羰基化底物,分为空白对照组、对照组及低、高浓度组(以HSYA 0.1 mmol/L、1 mmol/L干预).以2、4-二硝基苯肼(2、4-dinitrophenylhydrazine,DNPH)法检测蛋白质羰基化水平.结果 ONOO-途径和heme/NaNO2/H2O2途径均可以增加脑组织匀浆中羰基蛋白的含量.HSYA预处理可浓度依赖性地降低ONOO-途径诱导的蛋白质羰基化水平,低、高浓度组羰基蛋白含量分别降低了26.13%、46.23%,差异有统计学意义(F =14.265,P<0.05).HSYA预处理对体外heme/NaNO2/H2O2途径诱导的蛋白质羰基化没有明显抑制作用,差异无统计学意义(P>0.05).结论 HSYA可剂量依赖性地抑制ONOO-途径在体外对脑组织蛋白的羰基化修饰;提示HSYA抑制蛋白质羰基化反应可能是其对抗脑血管疾病与神经系统变性疾病的分子机制之一.%Objective To study the effects of Hydroxysafflor yellow A(HSYA)on brain protein carbonylation induced by peroxynitrite(ONOO-)and heme/NaNO2/H2O2 system in vitro.Methods To simulate in vivo peroxynitrite(ONOO-)and heme/NaNO2/H2O2 system-induced carbonylative pathway,with brain protein for nitrification substrates,divided into blank group,control group and HSYA intervention groups(doses of 0.1 and 1 mM,respectively).2,4-dinitrophenylhydrazine(DNPH)was used for the quantification of protein carbonylation.Results The treatment of brain tissue with ONOO-or heme/NaNO2/H2O2 resulted in the significant formation of carbonyl groups.HSYA,at doses of 0.1 and 1 mM,dose-dependently decreased ONOO-induced protein carbonylation by 26.13% and 46.23% respectively,the difference was statistically significant(F=14.625,P<0.05).However,HSYA was relatively ineffective in protecting brain tissue from heme/NaNO2/H2O2-induced oxidative damage,the difference was not statistically significant(P>0.05).Conclusion HSYA could dose-dependently inhibit brain protein carbonylative modification induced by ONOO-system in vitro,which may be one of the molecular mechanisms to against cerebrovascular and neurodegenerative diseases.