人参提取物对过氧化氢致人神经母瘤细胞损伤的保护作用及机制研究

摘要

目的 观察人参提取物对过氧化氢致人神经母瘤细胞(SH-SY5Y)损伤的影响并探讨其作用机制.方法 采用MTT法确定人参二醇总皂苷(PDS)、人参总皂苷(GS)、人参非皂苷(GN)的最佳给药浓度.将处于对数生长期的SY5Y细胞加入过氧化氢(终浓度为300μmol/L)孵育4h造成细胞损伤模型,然后分为模型对照组、PDS给药组、GS给药组、GN给药组,另设空白对照组加入等体积细胞培养液.采用MTT方法检测细胞活力,Western blot法检测Nrf2、Keap1、Bcl-2、Bax、Caspase3、Caspase9蛋白表达情况.结果 与空白对照组相比,模型组对照组SH-SY5Y细胞存活率明显降低(P＜0.01);与模型对照组比较,PDS给药组、GS给药组、GN给药组细胞存活率均升高,确定1、10、240 μg/mL为PDS给药组、GS给药组、GN给药组实验浓度(P＜0.01).与空白对照组比较,模型对照组SH-SY5Y中Keap1、Bax、Caspase3、Caspase9蛋白表达水平明显增加(P＜ 0.05或P＜0.01),Bcl-2、Nrf-2表达明显降低(P＜0.01);与模型组对照组比较,PDS给药组、GS给药组SH-SY5Y中Keap1、Bax、Caspase3、Caspase9蛋白表达水平明显降低(P＜ 0.05或P＜ 0.01),Bcl-2、Nrf-2表达明显增高(P＜0.01).结论 人参提取物中PDS与GS对过氧化氢致人神经母瘤细胞SY5Y细胞损伤具有保护作用,其机制可能与通过激活SH-SY5Y细胞内Nrf-2通路抑制氧化应激、降低其凋亡发生有关.%Objective To observe the influence of ginseng extract on the injury induced by hydrogen peroxide in human neuroblastoma cells (SH-SY5Y) and explore its mechanism.Methods The optimal concentrations of panaxadiol saponins (PDS),ginsenoside(GS) and ginseng non-saponin(GN) were determined by the MTT assay.SY5Y cells in the logarithmic growth phase were supplemented with hydrogen peroxide (final concentration of 300 μmol/L) and were incubated for 4 h.The cell injury model was established.SY5Y cells were divided into model control group,PDS group,GS group and GN group.A blank control group was added to an equal volume of cell culture fluid.MTT assay was used to detect the cell viability.The protein expressions of Nrf2,Keapl,Bcl-2,Bax,Caspase3 and Caspase9 were detected by Western blot.Results Compared with the blank control group,the survival rate of SH-SYSY cells in the model control group was significantly decreased (P ＜ 0.01).Compared with the model control group,the survival rates of PDS group,GS group and GN group increased,and the concentration of 1,10,240 μg/mL was selected as test concentration in PDS group,GS group and GN group (P ＜ 0.01).Compared with the blank control group,the protein levels of Keap1,Bax,Caspase3 and Caspase9 in SH-SY5Y of model control group were significantly increased (P ＜ 0.05 or P ＜ 0.01),and the protein levels of Bcl-2 and Nrf-2 were significantly decreased (P ＜ 0.01).Compared with the model control group,in PDS group and GS group the protein levels of Keap1,Bax,Caspase3 and Caspase9 were significantly decreased (P ＜ 0.05 or P ＜ 0.01),and the protein levels of Bcl-2 and Nrf-2 were significantly increased (P ＜ 0.01).Conclusion PDS and GS from ginseng extract can protect human SH-SY5Y cells from the hydrogen peroxide-induced damage,and the mechanism may inhibit the oxidative stress by activating the Nrf-2 pathway in SH-SY5Y cells,and reduce apoptosis.