人参总皂苷对过氧化氢诱导的红细胞氧化应激的抑制作用及机制研究

Study on the mechanism of total ginsenoside ginseng root inhibiting oxidative stress of red blood cells induced by hydrogen peroxide

目的探讨人参总皂苷(total ginsenoside ginseng root,TGGR)对过氧化氢(hydrogen peroxide,H_(2)O_(2))诱导的红细胞氧化应激的抑制作用及机制研究。方法利用70 mmol/L H_(2)O_(2)氧化诱导大鼠红细胞1 h,建立红细胞体外氧化应激损伤模型。将大鼠红细胞随机分为对照组、模型组、TGGR组(100μg/mL)。评价TGGR对H_(2)O_(2)诱导的红细胞的溶血率,凋亡率,红细胞形态,高铁血红蛋白(met-hemoglobin,Met-Hb)、游离血红蛋白(free-hemoglobin,F-Hb)、活性氧(reactive oxygen species,ROS)、丙二醛(malonaldehyde,MDA)、三磷酸腺苷(adenosine triphosphate,ATP)的水平,谷胱甘肽(glutataione,GSH)的含量,抗氧化酶、Na+/K+-ATP酶、Ca^(2+)/Mg^(2+)-ATP酶、糖酵解关键酶的活力,以及Band 3蛋白表达水平的影响。结果100μg/mL的TGGR能显著降低H_(2)O_(2)诱导的红细胞溶血率及凋亡率、降低Met-Hb和F-Hb水平(P<0.001,P<0.05),恢复红细胞形态,并显著提高GSH含量、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)的活力(P<0.001,P<0.01,P<0.05),显著降低ROS和MDA水平(P<0.01),发挥氧化损伤保护作用;TGGR可显著提高ATP水平和ATP酶活性(P<0.05,P<0.01),显著增强糖酵解关键酶活性,上调Band 3蛋白的表达,诱导糖酵解途径发挥抗氧化保护作用。结论TGGR通过上调Band 3蛋白提高红细胞糖酵解能力,恢复细胞内氧化平衡,从而抑制H_(2)O_(2)诱导的红细胞氧化应激。

Objective To investigate the protective and molecular mechanism of total ginsenoside ginseng root(TGGR)on oxidative stress injury of red blood cells induced by hydrogen peroxide(H_(2)O_(2)).Methods Red blood cells were induced by 70 mmol/L H_(2)O_(2) oxidation for 1 h to establish an oxidative stress injury model of red blood cells in vitro.The red blood cells of rats were stochastically divided into control group,model group and TGGR group(100μg/mL).Effects of TGGR on H_(2)O_(2)-induced red blood cells hemolysis rate,apoptosis rate,red blood cells morphology,and the levels of met-hemoglobin(Met-Hb),free-hemoglobin(F-Hb),reactive oxygen species(ROS),malonaldehyde(MDA),adenosine triphosphate(ATP),the content of glutathione(GSH),and the activities of antioxidant enzymes,Na^(+)/K^(+)-ATPase,Ca^(2+)/Mg^(2+)-ATPase,key glycolytic enzymes,and the expression of Band 3 protein were evaluated.Results The 100μg/mL TGGR could significantly reduce the H_(2)O_(2)-induced red blood cells hemolysis rate and apoptosis rate,significantly reduce the levels of Met-Hb and F-Hb(P<0.001,P<0.05),restore the morphology of red blood cells,and significantly increase the content of GSH,significantly improve the activities of superoxide dismutase(SOD)and catalase(CAT)(P<0.001,P<0.01,P<0.05),significantly reduce the levels of ROS and MDA to protect red blood cells against oxidative damage(P<0.01).TGGR could significantly increase the levels of ATP and ATPases activities(P<0.05,P<0.01),significantly enhance the activities of key enzymes in glycolysis,and up-regulate the expression of Band 3 protein to induce the glycolytic pathway to play an antioxidant protection role.Conclusion TGGR can improve the glycolysis ability of red blood cells by upregulating Band 3 protein,thereby restore the intracellular oxidative balance and inhibite the oxidative stress of red blood cells induced by H_(2)O_(2).