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4 h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SHSY5Y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、240μg/m L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 d 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-SY5Y 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 Keapl, 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 Keapl, 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 ceils 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.
出处
《中国医药导报》
CAS
2017年第30期13-17,共5页
China Medical Herald
基金
吉林省科技厅自然科学基金项目(20160101229JC)
吉林省教育厅"十三五"科学技术项目(JJKH20170713KJ
JJKH20170714KJ)
