姜黄素对SH-SY5Y细胞血红素加氧酶同工酶表达的影响

目的研究姜黄素在神经母细胞瘤细胞(SH-SY5Y)中对血红素加氧酶(HO)同工酶表达的影响,探讨姜黄素通过维持HO-1和HO-2的平衡对神经细胞的保护作用。方法体外培养人神经母细胞瘤细胞(SH-SY5Y细胞),以不同浓度的姜黄素(0、1.25、5.0、20μmol·L-1)处理SH-SY5Y细胞24h,及用浓度为5.0μmol·L-1的姜黄素分别处理SH-SY5Y细胞0、12、24、48h。利用荧光探针DCFH-DA和荧光分光光度计检测细胞内活性氧的水平。通过RT-PCR检测HO-1、HO-2mRNA的水平,Western blot检测HO-1、HO-2蛋白的表达。结果姜黄素作用于SH-SY5Y细胞后活性氧水平降低(P<0.05)。HO-1mRNA和蛋白的表达增加(P<0.05),而HO-2mRNA和蛋白的表达水平明显减弱(P<0.05),并呈浓度-时间依赖性。结论姜黄素通过抗氧化应激保护SH-SY5Y细胞,并促进SH-SY5Y细胞中HO-1的表达,下调HO-2的表达(P<0.05),姜黄素对HO-1和HO-2的这种反向调节,可能是姜黄素抗氧化应激、发挥神经细胞保护作用的一个机制。

The effect of two ways regulation of Curcumin on expression of HO isozymes in SH-SY5Y cells

Aim To investigate the effects of Curcumin on Heme oxygenase isozymes in SH-SY5Y cells and explore a new mechanism of Curcumin in neuroprotection. Methods The human SH-SY5Y cells were cultured in vitro and treated with Curcumin at 0,1.25,5.0,20 μmol·L-1 for 24 h,or with Curcumin at 5.0 μmol·L-1 for 0,12,24,and 48 h. The active oxygen was detected by fluorescent probe DCFH-DA and fluorospectro-photometer. RT-PCR was used to detect the expression of HO-1 and HO-2 mRNA. Western blot was performed to detect the levels of HO-1 and HO-2 protein.Results The results showed that Curcumin could inhibit the levels of active oxygen(P<0.05). HO-1 levels were increased in a dose-and time-dependent manner (P<0.05) after Curcumin treatment.Whereas,the levels of HO-2 mRNA and protein were gradually decreased(P<0.05). A negative correlation was found between HO-1 and HO-2. Conclusion Curcumin has the capability of anti-oxidative stress and regulated HO-1 and HO-2 bilaterally,which may be a new mechanism of the function of Curcumin on anti-oxidative stress.