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目的:观察丁苯酞预处理对心肌缺血再灌注损伤的保护作用以及磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路在此过程中的作用。方法:将大鼠心肌细胞(H9C2)随机分为假手术组(Sham组)、缺血再灌注组(I/R组)、丁苯酞预处理组(I/R＋NBP组)。通过氧气剥夺以及无糖培养基的更换模拟6 h缺血4 h复灌心肌缺血模型。通过细胞计数检测试剂盒(CCK-8)检测不同药物浓度细胞活性，探索丁苯酞的最佳药物浓度；比色法检测各组乳酸脱氢酶(LDH)以及丙二醛(MDA)含量；蛋白质印迹法(Western blot)检测PI3K、磷酸化蛋白激酶B(p-Akt)和Akt的蛋白表达水平；定量聚合酶链反应(qPCR)技术检测白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)在核糖核酸(mRNA)水平的表达。多组之间比较采用单因素方差分析，两组之间比较采用n t检验。n 结果:丁苯酞的最佳用药浓度为100 μmol/L；I/R组MDA、LDH的表达量高于Sham组[(172.03±14.99) nmol/L比(38.98±6.49) nmol/L、(195.04±14.50)%比(100.00±0.00)%，n t＝14.106、11.354，n P<0.05]；I/R组IL-1β、TNF-α的表达量高于Sham组(22.36±2.71比1.00±0.00、6.20±0.31比1.00±0.00，n t＝13.660、28.874，n P<0.05)；I/R组PI3K、p-Akt的表达量也高于Sham组(0.61±0.34比0.34±0.01、0.95±0.04比0.36±0.04，n t＝13.487、18.392，n P<0.05)，差异有统计学意义。I/R＋NBP组MDA、LDH的表达量低于I/R组[(56.98±4.79) nmol/L比(172.03±14.99) nmol/L、(140.34±4.44)%比(195.04±14.50)%，n t＝12.661、7.890，n P<0.05]；I/R＋NBP组IL-1β、TNF-α的表达量低于I/R组(6.66±0.60比22.36±2.71、1.76±0.11比6.20±0.31，n t＝9.798、23.352，n P<0.05)，差异有统计学意义；I/R＋NBP组PI3K、p-Akt的表达量高于I/R组(1.00±0.05比0.61±0.34、1.27±0.04比0.95±0.04，n t＝11.440、10.432，n P<0.05)，差异有统计学意义。n 结论:丁苯酞可以通过减少细胞损伤、抑制氧化及炎性反应进而减轻心肌缺血再灌注损伤，该过程可能有PI3K/Akt通路的参与。“,”Objective:To observe the protective effect of butylphthalide preconditioning on myocardial ischemia-reperfusion injury (IRI) and the role of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) pathway in this process.Methods:H9C2 cardiomyocyte cell line was purchased from Wuhan Punosi Biology Co., Ltd.. H9C2 cardiomyocytes were randomly divided into Sham group, IRI group, Butylphthalein preconditioning group (IRI+ NBP group). The model of myocardial IRI was simulated by oxygen deprivation and the replacement of glucose-free medium for 6 h ischemia and 4 h reperfusion. The optimal drug concentration of butylphthalide was chosen by cell counting kit (CCK-8). The contents of lactate dehydrogenase (LDH) and malondialdehyde (MDA) in each group were determined by colorimetry. Western blotting was used to detect the protein expression levels of PI3K, phosphorylated protein kinase B (p-Akt) and Akt. The quantitative polymerase chain reaction (qPCR) was used to detect the expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) mRNA. SPSS20.0 statistical software was used for statistical analysis. The numerical variables were expressed by mean±standard deviation (n Mean±n SD). One-way ANOVA was used for comparison among groups, and t-test was used for comparison between the two groups.n Results:The optimal concentration of butylphthalide was 100 μmol/L. The expression of MDA and LDH in IRI group was significantly higher than that in Sham group [(172.03±14.99) vs. (38.98±6.49) nmol/L, and (195.04±14.50)% vs. (100.00±0.00)%, n t=14.106, 11.354, n P<0.05]; The expression of IL-1β and TNF-α in IRI group was higher than that in Sham group (22.36±2.71 vs. 1.00±0.00, and 6.20±0.31 vs. 1.00±0.00,n t=13.660, 28.874, n P<0.05); The expression of PI3K and p-Akt in IRI group was higher than that in Sham group (0.61±0.34 vs. 0.34±0.01, and 0.95±0.04 vs. 0.36±0.04,n t=13.487, 18.392, n P<0.05). The expression of MDA and LDH in IRI + NBP group was lower than that in IRI group [(56.98±4.79) nmol/L vs. (172.03±14.99) nmol/L, and (140.34±4.44)% vs. (195.04±14.50)%,n t=12.661, 7.890, n P<0.05]; The expression of IL-1β and TNF-α in IRI + NBP group was lower than that in IRI group [(6.66±0.60) vs. 22.36±2.71, and (1.76±0.11) vs. (6.20±0.31),n t=9.798, 23.352, n P<0.05]; The expression of PI3K and p-Akt in IRI + NBP group was higher than that in IRI group [(1.00±0.05) vs. (0.61±0.34), and (1.27±0.04) vs. (0.95±0.04),n t=11.440, 10.432, n P<0.05].n Conclusion:Butylphthalide can alleviate myocardial IRI by reducing cell injury, inhibiting oxidation and inflammation, which may be involved in the PI3K/Akt signal pathway.