Heme oxygenase-1 (HO-1) protects human lens epithelial cells (SRA01/04) against hydrogen peroxide (H2O2)-induced oxidative stress and apoptosis

作者：Ma, Tianju; Chen, Tingjun; Li, Peng; Ye, Zi; Zhai, Wei; Jia, Liang; Chen, Wenqian; Sun, Ang; Huang, Yang; Wei, Shihui^*; Li, Zhaohui^*

来源：Experimental Eye Research, 2016, 146: 318-329.

DOI：10.1016/j.exer.2016.02.013

• Cataract
• Human lens epithelial cells (SRA01/04)
• Heme Oxygenase-1 (HO-1)
• Oxidative stress
• Antioxidation
• Apoptosis

Objectives: This study aimed to investigate the protective role of heme oxygenase-1 (HO-1) in H2O2-induced oxidative stress and apoptosis in human lens epithelial cells (hLEC; SRA01/04). @@@ Methods: SRA01/04 cells were exposed to a hydrogen peroxide (H2O2) concentration gradient and inducers of HO-1 such as cobalt protoporphyrin (CoPP) and zinc protoporphyrin (ZnPP), respectively. In addition, an RNA silencing experiment was conducted to investigate the HO-1 function in this study. A Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability. Western blot and ELISA were used to detect the level of HO-1 expression. In our study, hLECs were exposed to 400 mu M hydrogen peroxide (H2O2) for 24 h with or without pretreatment with 10 mu M CoPP or 10 mu M ZnPP, respectively. Double immunofluorescence staining was used for cell identification and the qualitative expression of HO-1. Expression of HO-1 was monitored using Western blot and ELISA. Intracellular reactive oxygen species (ROS) were detected by flow cytometry analyses; commercial enzymatic kits were used to measure the levels of glutathione (GSH), as well as superoxide dismutase (SOD). The proportion of cell apoptosis was quantified by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The expression of caspase family (-8,-3) proteins was measured by Western blot analysis. @@@ Results: HO-1 significantly restored the cell viability under H2O2 injury via reducing the generation of ROS and increasing the levels of SOD and GSH activity. Moreover, HO-1 also inhibited H2O2-induced caspase-8 and caspase-3 proteins, thus significantly reducing the apoptosis of SRA01/04. An RNA silencing experiment demonstrated the increased resistance of LEC5 to oxidative stress specifically for increased levels of HO-1. @@@ Conclusions: These findings suggested that HO-1 protects human lens epithelial cells from H2O2-induced oxidant stress by upregulating antioxidant enzyme activity, reducing ROS generation, and thus inhibiting caspase family-dependent apoptosis.

• 出版日期2016-5
• 单位天津医科大学; 中国人民解放军总医院; 南开大学; 天津市眼科医院; 北京大学