乙酰唑胺对缺氧环境下H9C2细胞损伤的保护机制研究

Protection Mechanism of Acetazolamide to Hypoxia-induced H9C2 Cell Damage

目的研究乙酰唑胺对缺氧环境下H9C2细胞损伤的保护作用及其机制。方法采用三气培养箱建立H9C2大鼠心肌细胞缺氧损伤模型;MTT法检测乙酰唑胺作用于缺氧H9C2细胞的最佳浓度;显微镜下观察乙酰唑胺对缺氧H9C2细胞形态的影响;检测缺氧0、12、24、36、48 h后乙酰唑胺对缺氧H9C2细胞乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、肌酸激酶(CK)、丙二醛(MDA)含量;DCHF-DA染色法检测乙酰唑胺对缺氧H9C2细胞活性氧(ROS)产生量的影响;RT-PCR法检测低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、半胱氨酸蛋白酶(Caspase)-3 mRNA的表达。结果与缺氧模型组相比较,乙酰唑胺能够降低缺氧H9C2细胞ROS、MDA含量和LDH、CK活性,升高SOD活性,在基因水平上,乙酰唑胺能够明显升高HIF-1α、VEGF mRNA的表达,降低Caspase-3mRNA的表达(P<0.05,P<0.01)。结论乙酰唑胺能有效保护缺氧H9C2细胞。

Objective To investigate the protection mechanism of Acetazolamide to hypoxia-induced H9C2 cell damage. Methods Models of hypoxia-induced H9C2 myocardial cell damage in rats were established by three gasses incu- bator; the best concentration of Acetazolamide to hypoxia-induced H9C2 cell was detected by MTT method; morphological influence of Aeetazolamide to hypoxia-induced H9C2 cell was observed under microscope ; contents of lactate dehydrogenase （LDH） , superoxide dismutase （SOD） , ereatine kinase （CK） and Malondialdehyde （MDA） in Acetazolamide to hypoxia- induced H9C2 cell after anoxia for 0, 12, 24, 36 and 48 h were detected; the DCFH-DA staining was used to detect the re- active oxygen species （ROS） production of Acetazolamide to hypoxia-induced H9C2 cell; RT-PCT method was used to measure the expressions of hypoxia inducing factor -1α （HIF-1α）, vascular endothelial growth factor （VEGF） and cysteine proteinase （Caspase） -3 mRNA. Results MDA content and activities of LDH and CK obviously decreased by Acetazo- lamide than those in model group, while SOD activity increased. Expressions of HIF-1α and VEGF mRNA were significant- ly increased, and Caspase -3 mRNA expression were significantly decreased （P 〈 0. 05,P 〈 0. 01 ）. Conclusion Acetazo- lamide may effectively protect hypoxia-induced H9C2 cells.