竹节参总皂苷通过ERK/Nrf2通路对H2O2致SH-SY5Y细胞氧化损伤的保护作用研究

Protective Effect of Total Saponins of Panax japonicus Against H_2O_2-Induced Oxidative Stress Damage

目的:探讨竹节参总皂苷对H2O2致SH-SY5Y细胞氧化应激损伤的保护作用及其分子机制。方法:H2O2作用SH-SY5Y细胞建立氧化应激损伤模型,以不同浓度竹节参总皂苷(0.1、1、5、20μg/m L)预处理12 h,再加入H2O2继续培养12 h。通过MTT检测细胞活力;酶生化法检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;Western blotting检测Nrf2、p-ERK、p-P38蛋白表达;Real-Time PCR检测NQO1、GCLC的mRNA表达。结果:各浓度竹节参均能显著提高H2O2所导致的细胞活力降低、提高SOD活性、降低MDA含量,并能上调Nrf2、p-ERK蛋白及NQO1、GCLC mRNA表达。结论:竹节参总皂苷对H2O2致SH-SY5Y细胞氧化应激损伤具有保护作用,其作用机制可能与调控ERK/Nrf2通路,从而增强抗氧化基因表达有关。

Objective：To investigate the protective effects of total saponins of Panax japonicus （TSPJ）on H2O2-induced oxidative stress damage in SH-SYSY cells ,and to explore the underlying mechanism. Methods：SH-SYSY cells were incubated with 600 μmol/L H2O2 for 12 h, then treated with various concentrations of TSPJ （0. 1,1,5 and 20 μg/mL）for 12 h, and then incubated with 600μmol/ L H2O2 for 12 h. Cell viability was detected by MTr assay. Superoxide dicmutase （SOD）activities and malondialdehyde（MDA） contents were measured by biochemical assay kits. Protein levels of Nrf2, p-ERK, and p-P38 were detected by Western blotting. Levels of NQO1 and GCLC mRNA expression were determined by real-time PCR. Results：Compared with control group, H2 02 stimulated the decrease of cell viability and SOD activities as well as the increase of MDA contents, which were reversed by TSPJ treatment. Furthermore, TSPJ treatment up-regulated not only the decreased protein expressions of Nrf2 and p-ERK but also the decreased mRNA expression of NQO1 and GCLC. Conclusion：TSPJ can protect SH-SYSY cells from H2O2-induced oxidative stress damage. The mechanism may be related to up-regulating the phosphorylation of ERK thereby promoting the Nrf2 nuclear translocation and increasing the mRNA expression of an- tioxidant genes such as NQO1 and GCLC.