摘要
目的探讨没食子酸丙酯(propyl gallate,PG,赤芍801)对H2O2诱导人神经母细胞瘤细胞(SH-SY5Y)氧化损伤的影响及其作用机制。方法将SH-SY5Y细胞均分为对照组、H2O2模型组、PG1组、PG2组和PG3组,对照组常规加入DMEM/F12培养液培养;H2O2模型组加入H2O2200滋mol·L-1;PG1组加入PG 20滋mol·L-1+H2O2200滋mol·L-1;PG2组加入PG 40滋mol·L-1+H2O2200滋mol·L-1;PG3组加入PG 80滋mol·L-1+H2O2200滋mol·L-1。比较5组SH-SY5Y细胞的增殖活性,细胞培养物上清中8-OHdG,细胞内ROS、LDH释放量,Hoechst 33258染色结果、细胞周期变化情况及Caspase-3酶活性。结果 H2O2模型组细胞存活率显著低于对照组,细胞培养物上清中8-OHdG水平显著高于对照组,ROS水平显著高于对照组,LDH释放量显著高于对照组,具有非常显著性差异(P<0.01);PG1组、PG2组和PG3组细胞存活率均显著高于H2O2模型组,细胞培养物上清中8-OHdG水平均显著低于H2O2模型组,ROS水平均显著低于H2O2模型组,LDH释放量均显著低于H2O2模型组,差异具有统计学意义(P<0.05);PG可显著改善SH-SY5Y细胞凋亡情况,且具有剂量依赖关系;H2O2模型组G0/G1期百分率显著高于对照组,S期、G2/M期和PI均显著低于对照组,差异具有统计学意义(P<0.05);PG2组、PG3组G0/G1期百分率显著低于H2O2模型组,S期、G2/M期和PI均显著高于H2O2模型组,差异具有统计学意义(P<0.05);H2O2模型组Caspase-3酶活性显著高于对照组,差异有统计学意义(P<0.05);PG2组和PG3组Caspase-3酶活性显著低于H2O2模型组,差异有统计学意义(P<0.05)。结论 PG在一定剂量范围内对H2O2诱导的SH-SYSY神经细胞损伤具有明显的保护作用,其保护效应可能与清除ROS,减轻DNA氧化损伤,抑制线粒体通路介导的细胞凋亡有关。
Objective To explore the effects of SH-SY5Y cells induced by hydrogen peroxide H202 model group, PG1 group, PG2 group, propyl （ H2O2 and gallate （ PG ） ）. Methods from Radix Paeoniae Rubra on oxidative damage in SH-SY5Y cells were divided into control group, PG3 group. The control group were cultured with DMEM/F12 nutrient solution, H202 model group were cultured with DMEM/F12 nutrient solution and H202 200 μmol·L^-1. Based on the culture solution for H202 model group, PG1 group, PG2 group, and PG3 group were co-culture with PG 20, 40 and 80 μmol·L^-1 respectively. The proliferation activity of SH-SY5Y cells, the release amount of 8-OHdG, ROS, LDH in the cell culture supernatant, the dyeing results of Hoeehst 33258, the changes of cell cycle, and the enzyme activity of Caspase-3 were compared among the 5 groups. Results In HzOz model group, the survival rate of SH-SY5Y cells was lower, the levels of 8-OHdG and ROS and the release amount of LDH in the cell culturesupernatant were higher than the control group（P〈 0.01 ）. PG groups had higher survival rate of SH-SY5Y cells, and lower 8-OHdG, ROS and LDH levels than the H202 model group（P〈 0.05）. PG could improve the cell apoptosis of SH-SY5Y dose-dependently. The percentage of G0/G1 stage of the H202 model group was higher, while the percentages of S stage, G2/M stage and PI% were lower than the control group（P 〈 0.05）. PG2 and PG3 groups had lower percentage of G0/G1 stage, and higher percentages of S stage andG2/M stage and PI than the H202 model group （P 〈 0.05）. The activity of Caspase-3 of the H202 Model group was higher than the control group（P 〈 0.05）, and then was decreased in PG2 group, PG3 group（P〈 0.05 compared with H202 model group）. Conclusion It is indicated that PG has neuroprotection against H202 -induced oxidative damage in certain dose range, and the protective mechanism is probably related with the elimination of ROS, the relief of DNA oxidative damage, and the inhibition of the apoptosis mediated b
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2014年第5期582-585,共4页
Traditional Chinese Drug Research and Clinical Pharmacology
关键词
没食子酸丙酯
H2O2
SH-SY5Y细胞
氧化损伤
作用机制
Propyl gallate（PG） from Radix Paeoniae Rubra
Hydrogen peroxide（H_2O_2）
SH-SY5Y cells
Oxidativedamage
Therapeutic mechanism
