姜黄素通过诱导Nrf-2上调SH-SY5Y细胞中HO-1的表达

目的研究姜黄素(Curcumin)对于血红素加氧酶-1(heme oxygenase-1,HO-1)和核因子相关因子2(nuclear factor erythroid2-related factor2,Nrf-2)表达的影响,探讨Curcumin保护神经细胞的作用机制。方法 SH-SY5Y(人神经母细胞瘤)细胞用0、1.25、5.0、20.0μmol·L-1 Curcumin处理24h,以及用5.0μmol·L-1 Curcumin分别处理细胞0、12、24、48h,然后用RT-PCR和Western blot检测各组的mRNA和蛋白的表达,并在各个浓度组使用Nrf-2siRNA,Western blot检测转染Nrf-2siRNA后HO-1的表达情况。结果 Curcumin处理后,SH-SY5Y细胞中Nrf-2和HO-1的表达均明显增强,且呈时间和浓度依赖性(P<0.05),0、1.25、5.0、20.0μmol·L-1 Curcumin处理的细胞24h后HO-1表达与用0,1.25,5.0,20.0μmol·L-1 Curcumin和转染Nrf-2siRNA同时处理24h后的细胞HO-1表达相比明显下降(group 1 vs group 5,group 2 vs group 6,group 3 vsgroup 7,group 4 vs group 8,P<0.05)。结论 Curcumin通过Nrf-2诱导HO-1的表达作用,可能是其抑制神经细胞氧化应激、发挥保护作用的机制之一。

Curcumins upregulate expression of HO-1 via inducing Nrf-2 in SH-SY5Y cells

Aim To investigate the effect of Curcumin on the expression of HO-1 and Nrf-2,and explore the machanism of neuroprotection of Curcumin. Methods SH-SY5Y cells were treated with Curcumin at 0,1. 25, 5. 0,20. 0 μmol·L -1,or with Curcumin at 5. 0μmol· L -1 for 12,24,48 h for the time course assay. RT-PCR and Western blot assays were carried out to detect mRNA and protein expression of Nrf-2 and HO-1. Nrf-2 siRNA was used to dectect the expression of HO-1 with the absence of Nrf-2. Results RT-PCR and Westernblot results indicated that the mRNA and protein expression of HO-1 and Nrf-2 all increased in a dose-and time-dependent manner after Curcumin treatment in the SH-SY5Y cells ( P < 0. 05) ,and the expression of HO-1 protein sharply decreased in the SH-SY5Y cells treated with Curcumin at 0,1. 25,5. 0,20. 0 μmol·L -1 compared with Nrf-2 siRNA transfected cells co-treated with 0,1. 25,5. 0,20. 0 μmol·L -1 Curcumin at 24 hr ( P <0. 05) . Conclusion Upregulating HO-1 expres-sion via inducing Nrf-2 may be a mechanism of Curcumin in anti-oxidative stress and cytoprotection.