壳寡糖对三氧化二砷致大鼠肝细胞毒性的保护作用

Protective effect of chitosan oligosaccharide against the toxicity of arsenic trioxide toward Buffalo rat liver cells

为研究壳寡糖(COS)对三氧化二砷(ATO)致大鼠正常肝细胞(BRL-3A)毒性的保护作用,测定三氧化二砷和壳寡糖对BRL-3A细胞的增殖抑制率、三氧化二砷诱导壳寡糖预处理BRL-3A细胞的细胞存活率、细胞内乳酸脱氢酶(LDH)水平、丙二醛(MDA)、谷胱甘肽S-转移酶(GST)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和线粒体膜电位(MMP)的变化。结果表明,三氧化二砷对BRL-3A细胞的半数抑制浓度(IC_(50))为19.8μmol·L^(-1),壳寡糖浓度低于230μmol·L^(-1)时对细胞并未产生损伤。壳寡糖预处理BRL-3A细胞24 h,三氧化二砷再处理细胞24 h,通过检测以上指标表明壳寡糖预处理能够降低LDH水平至176.23%±5.87%,从而降低三氧化二砷引起的细胞毒性;壳寡糖可显著抑制三氧化二砷引起的脂质过氧化对机体造成的损伤作用,MDA相对释放含量降低至191.91%±2.89%;壳寡糖显著提高了抗氧化酶GST、SOD和CAT的活性,分别提高至86.20%±1.41%、90.09%±1.33%和61.29%±0.21%,提高细胞的抗氧化能力,降低三氧化二砷引起的细胞氧化应激损伤;壳寡糖预处理可减缓三氧化二砷引起的线粒体膜电位的降低,降低三氧化二砷对细胞的线粒体产生的伤害。壳寡糖对三氧化二砷引起的大鼠肝细胞的细胞毒性损伤起到了有效地保护作用。

To study the protective effect of chitosan oligosaccharide(COS)against the toxicity of arsenic trioxide(ATO)toward normal Buffalo rat liver cells(BRL-3A),we determined the proliferation inhibition rate of BRL-3A cells induced by arsenic trioxide and chitosan oligosaccharide,cell survival rate of BRL-3A cells pretreated with chitosan oligosaccharides induced by arsenic trioxide,lactate dehydrogenase(LDH)release,malondialdehyde(MDA)assay,glutathione s-transferase(GST)activity,superoxide dismutase(SOD)activity,catalase(CAT)activity and mitochondrial membrane potential(MMP).The results showed that the IC_(50) of ATO to BRL-3A cells was 19.8μmol·L^(-1),while COS did not cause damage when it was below 230μmol·L^(-1).BRL-3A cells were pretreated with COS for 24 h,and then ATO was applied to the cells for another 24 h.Through the detection of LDH,MDA,GST,SOD,CAT and MMP,COS pretreatment could reduce the level of LDH release to 176.23%±5.87%induced by ATO,thus reduced the cytotoxicity induced by ATO;COS could effectively inhibit the damage caused by ATO-induced lipid peroxidation,and relative release content of MDA decreased to 191.91%±2.89%;COS could significantly increase the activities of antioxidant enzymes GST,SOD and CAT,increased to 86.20%±1.41%,90.09%±1.33%and 61.29%±0.21%,respectively,improve the antioxidant capacity and defense ability of cells,and reduce the oxidative stress damage induced by ATO;COS pretreatment could inhibit the decrease of mitochondrial membrane potential induced by ATO and reduce the damage of ATO to mitochondria;COS pretreatment could retard the decrease of mitochondrial membrane potential induced by arsenic trioxide and reduce the mitochondrial damage caused by arsenic trioxide.Therefore,chitosan oligosaccharide has a certain protective effect on the toxic damage of BRL-3A induced by arsenic trioxide.