Melatonin could attenuate oxidative damage in PC12 cells caused by acrylamide
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目的 探讨褪黑素(melatonin,MT)能否减轻丙烯酰胺(ACR)对PC12细胞的氧化损伤作用.方法 研究ACR对PC12细胞存活率和氧化损伤的时间剂量-效应关系.MT干预试验分为对照组、ACR组、ACR+ MT组(50 μmoL/LMT)、ACR+ MT+褪黑素受体拮抗剂Luzindole组(50μ mol/L MT和0.2μmol/L Luzindole).干预方式有提前24 h,同时以及推后3h干预3种.检测细胞内的活性氧(ROS)水平和丙二醛(MDA)含量.结果 2.5和5 mmo1/L ACR处理24 h能显著降低细胞存活率,与对照组比较差异有统计学意义(P<0.01).1.25 mmol/L ACR在6和12 h ROS水平较对照组明显升高(P <0.05);2.5 mmol/L ACR在1和6 h ROS水平明显高于对照组(P<0.05);5 mmol/L ACR在1 h ROS水平明显升高(P<0.05),在6和12 h MDA含量显著高于对照组(P<0.01).干预试验显示,MT提前24 h干预,ACR+ MT组细胞内ROS水平和MDA含量均显著低于ACR组(P<0.01),ACR+ MT+ Luzindole组细胞内ROS水平较ACR+ MT组显著升高(P<0.01);MT同时干预,ACR+ MT组细胞内ROS水平显著低于ACR组(P<0.01),ACR+ MT+ Luzindole 组ROS水平较ACR+ MT组显著升高(P<0.01),但对MDA含量未见明显影响;MT推后干预,细胞内ROS水平和MDA含量均未见显著影响.结论 3种不同干预方式中,提前24 h MT干预能显著减轻ACR导致的PC12细胞的氧化损伤作用.褪黑素的作用机制可能与MT受体有关.
Objective To study the influence of Melatonin(MT) on acrylamide-caused oxidative damage in PC12 cells.Methods 5 time points(1 h,3 h,6 h,12 h,24 h) and 3 doses of ACR (1.25 mmol/L,2.5 mmol/L,5 mmol/L) were designed to investigate the cell viability and toxicity.The cells were divided into four groups,control group,ACR group,ACR + MT group,ACR + MT + Luzindole group(50 μ mol/L MT was added either 24 h before or 3 h after or simultaneously with ACR treatment).The ROS level and MDA content were measured.Results The cell viability was significantly lower than the control group with ACR treatment for 24 h in 2.5 mmol/L and 5 mmol/L ACR groups(P <0.01).Compared with the control group,the ROS level in PC12 cells roses significantly at 6h and 12 h in 1.25 mmol/L ACR group(P <0.05).The ROS level at 1 and 6 h in 2.5 mmol/L ACR group was significantly higher than that in the control group(P < 0.05).The ROS level at 1 h and MDA content at 6 and 12 h in 5 mmol/L ACR group were obviously higher than that in the control group(P <0.05).In addition,compared with ACR group,the ROS level and MDA content were significantly lower in ACR + MT group when MT was added 24 h before ACR treatment(P < 0.01),and the ROS level was also significantly lower in ACR + MT group when MT and ACR were simultaneously added(P < 0.01).This kind of effect of MT could be attenuated by Luzindole.No obvious change was seen between ACR + MT group and ACR group when MT was added 24 h after ACR treatment.Conclusion When added 24 h before ACR treatment,MT was found to reduce the enhancement of ROS level and the MDA content in PC12 cells.This kind of reduction might depend on MT receptor.