和厚朴酚对过氧化氢诱导的人神经母细胞瘤细胞损伤的保护作用及机制研究

Protective effect and mechanism of honokiol on human neuroblastoma cell injury induced by hydrogen peroxide

目的观察和厚朴酚(HNK)对过氧化氢(H2O2)诱导的人来源神经母细胞瘤细胞(SH-SY5Y)损伤的保护现象,并探究其可能机制。方法用400μmol/L H2O2处理SH-SY5Y细胞24 h,建立细胞损伤模型。保护组用HNK预孵SH-SY5Y细胞6 h后,加入400μmol/L H2O2共同作用24 h,探讨HNK的作用。采用CCK-8比色法测定细胞活力;流式细胞术测定细胞凋亡;Western blot法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3表达,以及内质网应激(ERS)相关蛋白BIP、CHOP及Caspase-12表达。结果HNK增加H2O2作用下SH-SY5Y细胞活力(P<0.01),降低细胞凋亡率,增高Bcl-2/Bax比值,降低Caspase-3(P<0.05)、BIP、CHOP及Caspase-12蛋白表达(P<0.01)。结论HNK通过抑制ERS相关的凋亡保护H2O2对SH-SY5Y细胞的损伤作用。

Objective To observe the protective effect of honokiol(HNK) on hydrogen peroxide(H2O2)-induced injury of human neuroblastoma cells(SH-SY5 Y) and explore its possible mechanism.Methods SH-SY5 Y cells were treated with 400 μmol/L H2O2 for 24 hours to establish a cell injury model. In the protection group, SH-SY5 Y cells were preincubated with HNK for 6 hours, and then treated with 400 μmol/L H2O2 for 24 hours. The activity of SH-SY5 Y cells was determined by CCK8 colorimetry. Apoptosis of SH-SY5 Y cells was determined by flow cytometry. The protein expressions of cytoplasmic Bcl-2, Bax, caspase-3, BIP, CHOP and caspase-12 were detected by Western blot.Results HNK promoted the activity of SH-SY5 Y cells treated with H2O2(P<0.01), decreased cell apoptosis rate, increased Bcl-2/Bax ratio, and decreased protein expressions of Caspase-3(P<0.05), BIP, CHOP and Caspase-12(P<0.01).Conclusion HNK protects SH-SY5 Y cells from H2O2-induced damage by inhibiting ERS-associated cell apoptosis.