The effect and underlying mechanism of Timosaponin B-II on RGC-5 necroptosis induced by hydrogen peroxide

BMC Complementary and Alternative Medicine

Kun Xiong¹  Ju-Fang Huang¹  Ruo-Fei Ma³  Shuang Chen¹  Wei Ding¹  Li-Xiang Xue²  Lei Shang¹  San-Hong Jiang¹ 

[1] Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha, Hunan 410013, China;Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100191, China;Xiangya School of Medicine, Central South University, Changsha, Hunan 410013, China

关键词: Oxidative stress;    Necroptosis;    TNF-α;    Timosaponin B-II;    Retinal ganglion cells-5;   

Others  :  1085269
DOI  :  10.1186/1472-6882-14-459

 received in 2014-08-22, accepted in 2014-11-26,  发布年份 2014

【 摘 要 】

Background

Necroptosis is an important mode of cell death, which is due to oxidant stress accumulation. Our previous study indicated that oxidant stresses could be reduced by Timosaponin B-II (TBII), a kind of Chinese herb RhizomaAnemarrhenae monomer extraction. We wonder the possible effect of Timosaponin B-II, whether it can protect cells from necroptosis via reducing the oxidant stress, in RGC-5 following hydrogen peroxide (H₂O₂) insult.

Methods

RGC-5 cells were grown in DMEM, the model group was exposed in H₂O₂ with the concentration of 300 μM, and the experimental group was pre-treated with Timosaponin B-II at different concentrations (1 μM, 10 μM, 100 μM and 1000 μM) for 24 hrs. MTT assay was carried out to measure the cytotoxicity of H₂O₂, MDA concentration assay was executed to evaluate the degree of oxidative stress, TNF-α ELISA Assay was used to measure the concentration of TNF-α, finally, the degree of necrosis were analyzed using flow cytometry.

Results

We first constructed the cell injury model of necroptosis in RGC-5 upon H₂O₂ exposure. Morphological observation and MTT assay were used to evaluate the degree of RGC-5 death. MDA assay were carried out to describe the degree of oxidant stress. Annexin V/PI staining was used to detect necroptotic cells pre-treated with or without Timosaponin B-II following H₂O₂ injury. TNF-α ELISA was carried out to detect the TNF-α accumulation in RGC-5. Upon using Timosaponin B-II with concentration of 100 μM, the percentage of cell viability was increased from 50% to 75%, and the necrosis of cells was reduced from 35% to 20% comparing with H₂O₂ injury group. Oxidant stress and TNF-α was reduced upon injury which decreased the ratio of RGC-5 necroptosis.

Conclusion

Our study found out that Timosaponin B-II might reduce necroptosis via inhibition of ROS and TNF-α accumulation in RGC-5 following H₂O₂ injury.

【 授权许可】

   
2014 Jiang et al.; licensee BioMed Central Ltd.

【 图 表 】

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