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、50 μg/mL 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-eNOS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-eNOS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eNOS磷酸化发挥作用。

Objective To investigate the protective effects and mechanism of the extract from Cajanus cajan leaves (ECCL) against H₂O₂-induced oxidative injury in H9c2 cardiomyoblasts. Methods A model of H₂O₂-induced injury in H9c2 cardiomyoblasts was established. Cell viability was determined colorimetrically by MTT assay. The supernates and cells were collected, respectively, after the different treatments for measuring the LDH, MDA, and SOD levels with the corresponding detection kit according to the manufacturer's instructions. Western blotting was performed to exam the expression of p-Akt and p-eNOS in H9c2 cells respectively. Results Compared with H₂O₂ group, the cell viability was increased significantly in ECCL + H₂O₂ groups (P < 0.01). The activity of LDH in the culture medium was decreased significantly (P < 0.05). The content of MDA in the culture medium was decreased significantly (P < 0.05). The activity of SOD was increased significantly (P < 0.01). Treated with ECCL, the expressions of p-Akt and p-eNOS in H9c2 cells injured from H₂O₂ were increased significantly (P < 0.01), When LY294002 (inhibitor of PI3K) was added, the effects of ECCL were cancelled. Conclusion ECCL protects H9c2 cells against H₂O₂-induced oxidative injury partly through PI3K signaling pathway.

国家自然科学基金青年科学基金资助项目(81303262);山西中医学院博士科研启动基金;山西中医学院方药药效及其作用机制研究重点科技创新团队(20150401)