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目的研究木豆叶提取物(ECCL)对H2O2诱导的H9c2细胞氧化应激损伤的保护作用及其机制。方法建立H2O2诱导的H9c2细胞氧化损伤模型,于H2O2刺激前加入PI3K信号通路阻断剂LY294002(LY)预处理10 min,加入20、50μg/m L ECCL预处理24 h。MTT法检测细胞存活率,比色法测定上清液中乳酸脱氢酶(LDH)、丙二醛(MDA)水平和超氧化物歧化酶(SOD)的活性,Western blotting检测细胞内p-Akt、p-e NOS蛋白表达。结果与模型组比较,ECCL可明显增加H9c2细胞H2O2损伤后细胞存活率(P<0.01),增加SOD活力,降低MDA、LDH水平,增加p-Akt和p-e NOS蛋白表达(P<0.05、0.01),LY可阻断ECCL对H9c2的上述作用。结论 ECCL能够减轻H2O2所致的H9c2细胞损伤,可能是通过激活PI3K通路促进其下游因子Akt和e NOS磷酸化发挥作用。
Objective To investigate the protective effect and its mechanism of extract from Magnolia L on the oxidative stress injury induced by H2O2 in H9c2 cells. Methods H2O2-induced oxidative injury in H9c2 cells was established. Pretreatment with PI3K signaling pathway inhibitor LY294002 (LY) for 10 min before H2O2 stimulation and pretreatment with 20, 50 μg / mL ECCL for 24 h. Cell viability was measured by MTT assay. Lactate dehydrogenase (LDH), malondialdehyde (MDA) and superoxide dismutase (SOD) activity in supernatant were determined by colorimetric assay. The levels of p-Akt , Pe NOS protein expression. Results Compared with model group, ECCL significantly increased cell viability (P <0.01), increased SOD activity, decreased MDA and LDH levels, and increased p-Akt and pe NOS protein expression in H9c2 cells (P <0.05, 0.01) , LY can block the above effects of ECCL on H9c2. Conclusion ECCL can reduce H 2 O 2 -induced injury of H9c2 cells, which may be through the activation of PI3K pathway to promote its downstream factors Akt and e NOS phosphorylation.