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[目的]观察葡萄籽原花青素(grape seed proanthocyanidin,GSP)对过氧化氢(H2O2)致大鼠海马神经元损伤的保护作用。[方法]实验分为5组:阴性对照组(A组)、H2O2模型组(B组)、GSP低剂量组(C组,1.25mg/L)、GSP中剂量组(D组,2.50mg/L)和GSP高剂量组(E组,5.00mg/L)。采用原代细胞培养的方法,建立H2O2致海马细胞氧化损伤模型。观察各组细胞形态和超微结构;采用四甲基偶氮唑盐(MTT)比色法检测细胞存活率;化学比色法检测细胞内超氧化物歧化酶(SOD)活性、活性氧(ROS)和丙二醛(MDA)含量;TdT介导的dUTP缺口末端标记法(TUNEL法)检测细胞凋亡率。[结果]形态学观察C、D、E组均可减轻H2O2诱导的海马细胞氧化损伤。与B组相比,C、D、E组细胞存活率由31.97%分别上升到61.42%、68.47%和80.04%(P<0.01);细胞凋亡率由8.83%分别降低到3.50%、2.83%和2.00%(P<0.01);D、E组细胞内SOD含量明显增加(P<0.05或P<0.01),C、D、E组抑制ROS能力和减少MDA含量明显增加(P<0.01)。[结论]GSP对H2O2诱导的海马细胞氧化损伤有明显的保护作用,其作用机制可能与提高海马细胞的抗氧化能力以及减少凋亡率有关。
[Objective] To observe the protective effect of grape seed proanthocyanidin (GSP) on the damage of hippocampal neurons induced by hydrogen peroxide (H2O2) in rats. [Methods] The experiment was divided into 5 groups: negative control group (A group), H2O2 model group (B group), GSP low dose group (C group, 1.25 mg/L), GSP middle dose group (D group, 2.50 mg/ L) and GSP high dose group (E group, 5.00 mg/L). H2O2 induced oxidative damage model of hippocampal cells was established by primary cell culture. Cell morphology and ultrastructure were observed in each group; cell viability was measured by MTT colorimetric assay; superoxide dismutase (SOD) activity, reactive oxygen species (ROS) were detected by chemical colorimetry. ) and malondialdehyde (MDA) content; TdT mediated dUTP nick end labeling (TUNEL method) to detect the rate of apoptosis. [Results] Morphological observations of C, D and E groups all reduced H2O2-induced hippocampal oxidative damage. Compared with group B, the survival rate of C, D and E groups increased from 31.97% to 61.42%, 68.47% and 80.04% respectively (P<0.01); the apoptosis rate decreased from 8.83% to 3.50% and 2.83% respectively. And 2.00% (P <0.01); D, E group intracellular SOD content increased significantly (P <0.05 or P <0.01), C, D, E group inhibit ROS ability and reduce the MDA content increased significantly (P <0.01). [Conclusion] GSP has obvious protective effect on H2O2-induced oxidative damage of hippocampus, and its mechanism may be related to the increase of anti-oxidation ability and decrease of apoptosis rate of hippocampal cells.