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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(50μmol/L MT)、ACR+MT+褪黑素受体拮抗剂Luzindole组(50μmol/L MT和0.2μmol/L Luzindole)。干预方式有提前24 h,同时以及推后3 h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 mmol/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 investigate whether melatonin (MT) can reduce the oxidative damage of PC12 cells induced by acrylamide (ACR). Methods The time-dose-response relationship of ACR on PC12 cell viability and oxidative damage was studied. MT intervention experiments were divided into control group, ACR group, ACR + MT group (50μmol / L MT), ACR + MT + melatonin receptor antagonist Luzindole group (50μmol / L MT and 0.2μmol / L Luzindole). Intervention was 24 h ahead of time, and 3 h interventions at the same time and 3 h later. The levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in cells were measured. Results 2.5 and 5 mmol / L ACR treatment for 24 h significantly reduced cell viability, compared with the control group, the difference was statistically significant (P <0.01). The ROS level of 1.25 mmol / L ACR at 6 and 12 h was significantly higher than that of the control group (P <0.05). The ROS level at 2.5 mmol / L ACR at 1 and 6 h was significantly higher than that of the control group (P <0.05) The level of ROS in LACR was significantly increased at 1 h (P <0.05), and at 6 and 12 h, MDA content was significantly higher than that in control group (P <0.01). The results of intervention showed that the level of ROS and MDA in ACR + MT group was significantly lower than that in ACR group (P <0.01), and the level of ROS in ACR + MT + Luzindole group was significantly higher than that in ACR + MT group (P <0.01). The level of ROS in ACR + MT group was significantly lower than that in ACR group (P <0.01), while the level of ROS in ACR + MT + Luzindole group was significantly higher than that in ACR + MT group ), But no significant effect on MDA content; MT push post-intervention, intracellular ROS levels and MDA content were not significantly affected. Conclusions Among the three different interventions, 24 h MT pretreatment significantly reduced the oxidative damage induced by ACR in PC12 cells. The mechanism of action of melatonin may be related to the MT receptor.