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枸杞多糖保护人晶状体上皮细胞免受氧化应激诱导的凋亡和衰老。

Lycium barbarum polysaccharides protect human lens epithelial cells against oxidative stress-induced apoptosis and senescence.

作者信息

Qi Bing, Ji Qingshan, Wen Yuechun, Liu Lian, Guo Xiaoling, Hou Guanghui, Wang Guifang, Zhong Jingxiang

机构信息

Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China.

Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, Anhui, China.

出版信息

PLoS One. 2014 Oct 15;9(10):e110275. doi: 10.1371/journal.pone.0110275. eCollection 2014.

Abstract

OBJECTIVES

We aimed to investigate the protective effect of Lycium barbarum polysaccharides (LBPs) against oxidative stress-induced apoptosis and senescence in human lens epithelial cells.

METHODS

To study apoptosis, SRA01/04 cells, a human lens epithelial cell lines, were exposed to 200 µM hydrogen peroxide (H2O2) for 24 h with or without pretreatment with LBPs. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis, intracellular reactive oxygen species (ROS), and the loss of mitochondria membrane potential (Δψm) were detected by flow cytometric analyses. Expression levels of Bcl-2 and Bax proteins were measured by western blot analysis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) were quantized using commercial enzymatic kits according to the manufacturer's instructions. To study senescence, SRA01/04 cells were pre-incubated with LBPs and all cells were then exposed to 100 µM H2O2 for 96 h. Cellular senescence was assessed by morphologic examination and senescence-associated β-galactosidase (SA-β-gal) staining.

RESULTS

LBPs significantly reduced H2O2-induced cell apoptosis, the generation of ROS, the loss of Δψm, and the levels of MDA. LBPs also inhibited H2O2-induced downregulated Bcl-2 and upregulated Bax proteins and increased the levels of SOD and GSH enzyme activity. Moreover, LBPs significantly attenuated H2O2-induced cellular senescence.

CONCLUSIONS

These findings suggested that LBPs protect human lens epithelial cells from H2O2-induced apoptosis by modulating the generation of ROS, loss of Δψm, Bcl-2 family, and antioxidant enzyme activity and attenuating cellular senescence.

摘要

目的

我们旨在研究枸杞多糖(LBPs)对氧化应激诱导的人晶状体上皮细胞凋亡和衰老的保护作用。

方法

为研究细胞凋亡,人晶状体上皮细胞系SRA01/04细胞在有或无LBPs预处理的情况下,暴露于200μM过氧化氢(H2O2)中24小时。使用细胞计数试剂盒-8(CCK-8)测定法测量细胞活力。通过流式细胞术分析检测细胞凋亡、细胞内活性氧(ROS)以及线粒体膜电位(Δψm)的丧失。通过蛋白质印迹分析测量Bcl-2和Bax蛋白的表达水平。根据制造商的说明,使用商业酶试剂盒对丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)的水平进行定量。为研究细胞衰老,SRA01/04细胞先用LBPs预孵育,然后所有细胞暴露于100μM H2O2中96小时。通过形态学检查和衰老相关β-半乳糖苷酶(SA-β-gal)染色评估细胞衰老。

结果

LBPs显著降低了H2O2诱导的细胞凋亡、ROS的产生、Δψm的丧失以及MDA的水平。LBPs还抑制了H2O2诱导的Bcl-2下调和Bax蛋白上调,并增加了SOD和GSH酶活性水平。此外,LBPs显著减轻了H2O2诱导的细胞衰老。

结论

这些发现表明,LBPs通过调节ROS的产生、Δψm的丧失、Bcl-2家族和抗氧化酶活性以及减轻细胞衰老,保护人晶状体上皮细胞免受H2O2诱导的凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c0f7/4198253/11eecfdc7f2d/pone.0110275.g001.jpg

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