丁苯酞对缺氧缺糖/复氧复糖PC12细胞损伤的保护作用

Protective effects of butylphthalein on oxygen-glucose deprivation and reintroduction induced PC12 cells injury

目的探究丁苯酞对缺氧缺糖/复氧复糖PC12细胞损伤的保护作用和机制。方法将PC12细胞随机分为对照组(常规培养)、模型组[缺氧缺糖/复氧复糖(OGD/R)]和实验组(10μmol·L^(-1)丁苯酞)。然后,将NC inhibitor和miR-146b-5p inhibitor分别转染至实验组细胞中,分别命名为转染组-1和转染组-2。用酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-β)含量;试剂盒检测活性氧(ROS)和超氧化物歧化酶(SOD)含量;用蛋白免疫印迹法检测肿瘤坏死因子受体相关因子6(TRAF6)蛋白表达。结果对照组、模型组和实验组的TNF-α含量分别为(53.31±4.71),(115.10±10.33)和(82.13±4.83)pg·m L^(-1);IL^(-1)β含量分别为(62.17±4.88)(97.33±7.96)和(74.42±5.33)pg·m L^(-1);ROS活性分别为(21.12±2.11),(47.62±2.98)和(32.01±3.15)U·m L^(-1);SOD活性分别为(60.42±3.33),(44.78±2.69)和(68.01±5.12)U·m L^(-1);TRAF6蛋白相对表达量分别为1.03±0.11,2.92±0.32和1.93±0.20。上述指标,模型组与对照组相比,实验组与模型组相比,差异均有统计学意义(均P<0.05)。转染组-1和转染组-2中TRAF6相对表达量分别为1.01±0.10和3.12±0.29,两组比较差异有统计学意义(均P<0.05)。结论丁苯酞通过miR-146b-5p/TRAF6轴抑制炎症反应和氧化应激反应减轻缺氧缺糖/复氧复糖诱导的PC12细胞损伤。

ObjectiveTo investigate the protective effects of butylphthalein on oxygen-glucose deprivation and reintroduction induced PC12 cells injury.Methods PC12 cells were randomly divided into control group(normal culture),model group[oxygen-glucose deprivation and reintroduction(OGD/R)]and test group(10μmol·L^(-1)butylphthalein),and the test group were transfected with NC inhibitor or miR-146b-5p inhibitor,named transfection-1 group and transfection-2 group.Enzyme linked immunosorbent assay was used to detect the content of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL^(-1)β).The kit was used to detect the content of reactive oxygen species(ROS)and superoxide dismutase(SOD).The expression of tumor necrosis factor receptor-associated factor 6(TRAF6)protein was detected by Western blotting.Results In the control group,model group and test group,the content of TNF-αwere(53.31±4.71),(115.10±10.33)and(82.13±4.83)pg·m L^(-1),respectively;the content of IL^(-1)βwere(62.17±4.88),(97.33±7.96)and(74.42±5.33)pg·m L^(-1),respectively;ROS activity were(21.12±2.11),(47.62±2.98)and(32.01±3.15)U·m L^(-1),respectively;SOD activity were(60.42±3.33),(44.78±2.69)and(68.01±5.12)U·m L^(-1),respectively;the relative expressions of TRAF6 were 1.03±0.11,2.92±0.32 and 1.93±0.20.In terms of the above indicators,compared between the model group and the control group,the test group and the model group,the differences were statistically significant(P<0.05).The relative expressions of TRAF6 in the transfection-1 group and transfection-2 group were 1.01±0.10 and 3.12±0.29.The differences between two groups were statistically significant(P<0.05).Conclusion Butylphthalein inhibits inflammation and oxidative stress through the miR-146b-5p/TRAF6 axis to play a protective role in oxygen-glucose deprivation and reintroduction induced PC12 cells injury.