铁皮石斛叶、花对H_(2)O_(2)诱导HaCaT细胞氧化损伤的保护及LC-MS分析

Protective Effects of Dendrobium officinale leaf and flower on Hydrogen Peroxide-induced Oxidative Damage of HaCaT Cells and LC-MS Analysis

目的研究铁皮石斛叶、花大孔树脂醇洗脱部位对H_(2)O_(2)诱导HaCaT细胞氧化应激损伤的保护作用,为其开发利用提供科学依据。方法采用MTT法检测H_(2)O_(2)诱导的氧化应激损伤性HaCaT细胞的存活率,试剂盒羟胺法和比色法分别检测HaCaT细胞内SOD和GSH-Px活性;运用UPLC-Q-TOF-MS/MS分析其化学成分。结果与模型组比较,铁皮石斛叶、花大孔树脂醇洗脱部位均能显著提高H_(2)O_(2)诱导的HaCaT细胞的存活率以及细胞中SOD和GSH-Px活性(P<0.05)。分别从铁皮石斛叶、花大孔树脂醇洗脱部位指认19个、16个化学成分,叶、花中黄酮苷类成分均为11个,为其主要组成成分。结论铁皮石斛叶、花大孔树脂醇洗脱部位对H_(2)O_(2)诱导的HaCaT细胞均具有保护作用,黄酮苷类成分可能为其活性物质基础。

Objective The protective effects of the alcohol-eluted fractions of Dendrobium officinale leaf and flower from macroporous resin on hydrogen peroxide-induced oxidative stress injury in HaCaT cells were researched to provide scientific basis for their development and utilization.Methods MTT assay was used for survival rate of H_(2)O_(2)-induced oxidative damage in HaCaT cells.Then,the levels of SOD and GSH-Px in HaCaT cells were also detected by commercial kit with the hydroxylamine and colorimetry methodology,respectively.In addition,the components of the alcohol-eluted fractions were identified by UPLC-Q-TOF-MS/MS.Results Compared with the model group,the survival rate,SOD and GSH-Px activity(P<0.05)of HaCaT cells,H_(2)O_(2)-induced oxidative damage,were significantly increased by the alcohol-eluted fractions of Dendrobium officinale leaf and flower from macroporous resin.There were 19 and 16 compounds identified from alcohol-eluted fractions of Dendrobium officinale leaf and flower,respectively.Of which,11 flavonoid glycosides were the major components both in leaf and flower of Dendrobium officinale.Conclusion The alcohol-eluted fractions of Dendrobium officinale leaf and flower from macroporous resin had protective effects on H_(2)O_(2)-induced oxidative stress HaCaT cells,and the flavonoid glycosides might be bioactive components.