CAJ | 학술논문

目的观察高良姜素对人A375黑素瘤细胞氧化损伤后细胞内Nrf2、γ-GCS基因表达的影响,探讨其抗氧化损伤的保护机制.方法以 700 μmol/L 过氧化氢(H2O2)作为外源性氧化剂,诱导人 A375 黑素瘤细胞处于氧化应激状态,建立细胞氧化损伤模型.实验分为正常组、模型组、阳性药组和高良姜素低、中、高剂量组,各给药组加入相应药物培养.MTT法检测细胞存活率,ELISA检测胞内活性氧簇(ROS)含量,RT-PCR检测Nrf2、γ-GCS基因表达.结果与正常组比较,模型组细胞存活率显著下降,ROS含量明显上升,Nrf2与γ-GCS表达量均明显下降(P<0.05,P<0.01);与模型组比较,各药物组细胞存活率升高,ROS含量降低,Nrf2和γ-GCS基因表达量显著升高(P<0.05,P<0.01).结论高良姜素可能是通过上调细胞Nrf2和γ-GCS的表达,促进Nrf2与抗氧化反应元件及其相关调节酶的结合,从而激活Nrf2信号通路达到对A375细胞氧化损伤的保护作用.
목적 관찰고량강소대인A375흑소류세포양화손상후세포내Nrf2、γ-GCS기인표체적영향,탐토기항양화손상적보호궤제.방법 이 700 μmol/L 과양화경(H2O2)작위외원성양화제,유도인 A375 흑소류세포처우양화응격상태,건립세포양화손상모형.실험분위정상조、모형조、양성약조화고량강소저、중、고제량조,각급약조가입상응약물배양.MTT법검측세포존활솔,ELISA검측포내활성양족(ROS)함량,RT-PCR검측Nrf2、γ-GCS기인표체.결과 여정상조비교,모형조세포존활솔현저하강,ROS함량명현상승,Nrf2여γ-GCS표체량균명현하강(P<0.05,P<0.01);여모형조비교,각약물조세포존활솔승고,ROS함량강저,Nrf2화γ-GCS기인표체량현저승고(P<0.05,P<0.01).결론 고량강소가능시통과상조세포Nrf2화γ-GCS적표체,촉진Nrf2여항양화반응원건급기상관조절매적결합,종이격활Nrf2신호통로체도대A375세포양화손상적보호작용.
Objective To investigate the effects of galangin on gene expressions of Nrf2 andγ-GCS in oxidative damage of A375 cell;To discuss its protective mechanism for anti-oxidative damage. Methods A375 melanoma cells were induced oxidative stress to establish oxidative damage model by 700μmol/L H2O2. The study was divided into normal group, model group, positive medicine group and high-, medium-, and low-dose galangin groups. All administration groups were given relevant medicine for cultivation. Cell viability was detected by MTT;ROS content was detected by ELISA;the gene expressions of Nrf2 andγ-GCS were detected by RT-PCR.Results Compared with normal group, cell viability decreased significantly;ROS content increased significantly;the gene expressions of Nrf2 andγ-GCS decreased significantly in the model group (P<0.05,P<0.01). Compared with model group, cell viability increased, ROS content decreased, the gene expressions of Nrf2 andγ-GCS increased significantly in all administration groups (P<0.05,P<0.01). Conclusion Galangin may activate Nrf2 signal path to realize the protective effect on A375 cellular oxidation damage through upregulating the expressions of Nrf2 andγ-GCS to promote the integration of Nrf2 and antioxidant response element and relevant regulatory enzymes.