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目的:探讨竹节香附素A对缺氧条件下人肝癌细胞株(HepG2)细胞缺氧诱导因子-1α(hypoxiainducible factor-1α,HIF-1α)基因和蛋白表达的影响。方法:取体外低氧环境下HepG2细胞进行不同干预:特定浓度(10μg/ml)竹节香附素A不同作用时间(0 h、6 h、12 h、24 h);不同浓度(0、5、10、20μg/ml)竹节香附素A特定作用时间(24 h)处理,通过半定量逆转录-聚合酶链反应(RT-PCR)和Western blot检测HepG2细胞HIF-1αmRNA和蛋白的表达变化情况。结果:体外低氧环境下HepG2细胞可见HIF-1αmRNA和蛋白强表达,特定浓度(10μg/ml)竹节香附素A可显著抑制HIF-1αmRNA和蛋白的表达水平,药物作用0 h、6 h、12 h、24 h时HIF-1αmRNA条带灰度值(A/A_0)分别为0.73±0.08、0.44±0.12、0.24±0.09、0.07±0.03,差异具有统计学意义(P<0.01);HIF-1α蛋白A/A_0比值分别为2.31±0.33、1.70±0.17、1.30±0.96、0.09±0.13,差异具有统计学意义(P<0.01)。不同浓度竹节香附素A特定作用时间(24 h)处理可显著抑制HIF-1αmRNA和蛋白的表达水平,药物浓度0、5、10、20μg/ml处理,HIF-1αmRNA A/A_0比值分别为0.65±0.07、0.43±0.08、0.19±0.03、0.08±0.02,差异具有统计学意义(P<0.01);HIF-1α蛋白A/A_0比值分别为0.84±0.08、0.39±0.06、0.28±0.04、0.08±0.03,差异具有统计学意义(P<0.01)。结论:竹节香附素A对缺氧条件下肝癌HepG2细胞HIF-1αmRNA和蛋白的表达具有抑制作用,并存在时间-剂量依赖性效应。
AIM: To investigate the effects of sarcandra fumigatus A on hypoxiainducible factor-1α (HIF-1α) gene and protein expression in human hepatoma cell line HepG2 under hypoxia. Methods: HepG2 cells were exposed to hypoxia in vitro for different periods of time (0 h, 6 h, 12 h, 24 h) and different concentrations (0, 5 , 10, 20μg / ml Scutellarein A for 24 h, the expression of HIF-1α mRNA and protein in HepG2 cells was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot Changes. RESULTS: HIF-1αmRNA and protein were strongly expressed in HepG2 cells in hypoxia environment. Sarcandra A significantly inhibited the expression of HIF-1αmRNA and protein at certain concentration (10μg / ml) HIF-1αmRNA gray level (A / A_0) were 0.73 ± 0.08,0.44 ± 0.12,0.24 ± 0.09,0.07 ± 0.03 at 12 h and 24 h, respectively, the difference was statistically significant (P <0.01); HIF 1α protein A / A_0 ratio were 2.31 ± 0.33,1.70 ± 0.17,1.30 ± 0.96,0.09 ± 0.13, the difference was statistically significant (P <0.01). The concentration of flavonoid A at different concentrations (24 h) could significantly inhibit the expression of HIF-1αmRNA and protein. The drug concentration of 0, 5, 10, 20μg / ml and HIF-1αmRNA A / A_0 ratio were 0.65 ± 0.07,0.43 ± 0.08,0.19 ± 0.03,0.08 ± 0.02, the difference was statistically significant (P <0.01); The protein A / A_0 ratio of HIF-1α were 0.84 ± 0.08,0.39 ± 0.06,0.28 ± 0.04,0.08 ± 0.03, the difference was statistically significant (P <0.01). CONCLUSION: Sarcaxin A can inhibit the expression of HIF-1αmRNA and protein in HepG2 cells under hypoxia condition in a time-and dose-dependent manner.