高良姜素对过氧化氢诱导的A375细胞氧化损伤后Nrf2、γ-GCS基因表达的影响

Effects of Galangin on Gene Expressions of Nrf2 and γ-GCS in Oxidative Damage of A375 Cell Induced by H_2O_2

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700μmol/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。

Objective To investigate the effects of galangin on gene expressions of Nrf2 and γ-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 700 μmol/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 and γ-GCS were detected by RT-PCR. Results Compared with normal group, cell viability decreased significantly;ROS content increased significantly;the gene expressions of Nrf2 and γ-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 and γ-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 and γ-GCS to promote the integration of Nrf2 and antioxidant response element and relevant regulatory enzymes.