黄芪甲苷对缺氧/复氧损伤H9c2心肌细胞的影响

Effects of astragaloside IV on hypoxia-reoxygenation injury of H9c2 cardiomyocytes

目的:观察黄芪甲苷对缺氧/复氧损伤H9c2心肌细胞干预作用。方法:H9c2心肌细胞培养,传代后随机分为5组,对照组、缺氧/复氧组、黄芪甲苷药物预处理组:培养液中加入终浓度分别为25、50和100μmol/L的黄芪甲苷预处理,12h后进行缺氧/复氧处理。分别收取细胞培养上清液,用酶联免疫吸附技术(Elisa)测定上清液中的白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α),并检测超氧化物歧化酶(SOD)、丙二醛(MDA)含量;采用RT-PCR技术检测NF-κB的基因表达。结果:黄芪甲苷(25、50、100μmol/L)预处理12h后可明显降低IL-6、TNF-α、MDA的含量,可明显提高SOD含量,并可以减低核转录因子NF-κB P65基因表达水平,并且具有一定的量效关系。结论:黄芪甲苷保护缺氧/复氧对H9c2心肌细胞损伤,通过提高SOD含量,降低MDA含量,发挥抗氧化作用,并且抑制炎性因子IL-6、TNF-α分泌,其机制与减低核转录因子NF-κB P65基因表达水平有关。

Objective： To observe the protective effect of Astragaloside IV on hypoxia /reoxygenation injury of H9c2 cardiomyocytes. Methods：H9c2 myocardial cells cultured were randomly divided into five groups： control group,hypoxia /reoxygenation group,pre-treatment group by respective final concentration of 25,50 and 100μmol /L of Astragaloside IV before hypoxia /reoxygenation. Cell supernatants were collected in each group,and the content of interleukin-6（ IL-6）,tumor necrosis factor-α（ TNF-α） which were detected by enzyme-linked immunosorbent assay,superoxide dismutase（ SOD） and malondialdehyde（ MDA） content were measured in each goup; NF-κB gene expression were detected by using RT-PCR technique. Results： the content of IL-6,TNF-α,MDA significantly reduced,the SOD content significantly increased,and the nuclear transcription factor NF-κB P65 gene expression levels reduced in pretreatment groups by Astragaloside（ 25,50,100μmol/L）,which have a certain dose-effect relationship. Conclusion： Astragaloside IV have the protective effects on hypoxia-reoxygenation injury of H9c2 cardiomyocytes,such as increasing SOD levels,decrease MDA content play antioxidant effect,inhibiting inflammatory cytokines IL-6,TNF-α secretion by reducing their NF-κB P65 gene expression levels.