抗辐灵活性成分对微波辐射致PC12细胞氧化应激损伤的保护作用研究

目的：观察抗辐灵活性成分黄芪总苷、赤芍总苷和丹参酮对微波辐射致PC12细胞氧化应激损伤的保护作用。方法采用神经生长因子（ nerve growth factor，NGF）诱导的PC12细胞为模型，分别给予1、3和9μg／ml黄芪总苷、赤芍总苷、丹参酮，作用48 h后经30 mW／cm2微波辐射6 min，于辐射后即刻采用倒置相差显微镜观察PC12细胞形态变化，四甲基偶氮唑盐（ methylthiazolyl tetrazolium，MTT）比色法测定细胞生存率，活性氧探针二氯二氢荧光素二乙酸酯（DCFH－DA）法测定细胞内活性氧（reactive oxygen species，ROS）水平，硫代巴比妥酸（thiobarbituric acid，TBA）法测定细胞脂质过氧化代谢产物丙二醛（ malonyldialdehyde，MDA）含量。结果3种活性成分作用后经微波辐射各组PC12细胞形态未见明显差异。辐射后6 h，辐射组PC12细胞较对照组（假辐射组）生存率明显降低（P＜0．01）；与辐射组相比，黄芪总苷（3μg／ml）给药组PC12细胞生存率明显提高（P＜0．01）。辐射后0和48 h，辐射组PC12细胞ROS、MDA水平明显升高（P＜0．01），与辐射组相比，各药物预处理组PC12细胞ROS、MDA水平均显著降低（P＜0．05）。结论抗辐灵3种活性成分能改善微波辐射对PC12细胞的损伤，其保护作用可能是通过清除氧自由基和减轻氧化应激损伤来实现的。

Protective effect of active ingredients of Kang Fu Ling on PC12 cells oxidative injury induced by microwave irradiation

Objective To observe the effect of the three active ingredients of a Chinese traditional medicine compound named Kang Fu Ling( KFL) against PC12 cells oxidative damage induced by microwave radiation.Methods PC12 cells were differentiated into neuros induced by nerve growth factor ( NGF ) .PC12 cells were incubated for 48 hours after astragalosides,total paeony glycoside and tanshinones were added at different concentrations (1, 3, or 9 μg/ml) .The cells in the control group were cultivated with the only medium of the same volume.Then, cells were irradiated with 30 mW/cm2 microwave for 6 minutes.The morphology of PC12 cells was observed under an inverted microscope soon before and after irradiation and the cell viability was measured by methylthiazolyl tetrazolium ( MTT) colorimetry.Reactive oxygen species ( ROS ) was determined using active oxygen probe 2′, 7′-dichlorodihyarofluolescen diacetde ( DCFH-DA ) while malonyldialdehyde(MDA) was measured in the homogenate of PC12 cells through thiobarbituric acid ( TBA) reactive substance assay.Results The cell morphology of each group showed no obvious difference.6 h after irradiation, the viability of irradiation control group measured by MTT declined apparently(P<0.01)compared with the normal control group.The 3 μg/ml astragalosides treatment group increased the viability of PC12 cells after microwave exposure ( P <0.01).The contents of ROS and MDA were increased after irradiation(P<0.01).However, in the three active ingredients of Kang Fu Ling treatment groups, both ROS and MDA were much lower than in irradiation control group.Conclusion Astragalosides, total paeony glycoside and tanshinones, which are the three active ingredients of Kang Fu Ling, all have protective effect against PC12 cell injury caused by microwave radiation,possibly by scavenging free radicals and reducing oxidative stress injury.