Bai Jie, Zheng Yi, Wang Gang, Liu Ping
Key Laboratory of Harbin Medical University Eye Center, Eye Hospital, First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Oxid Med Cell Longev. 2016;2016:5962832. doi: 10.1155/2016/5962832. Epub 2015 Nov 23.
Oxidative stress, as mediated by ROS, is a significant factor in initiating the development of age-associated cataracts; D-limonene is a common natural terpene with powerful antioxidative properties which occurs naturally in a wide variety of living organisms. It has been shown to have antioxidant effect; we found that D-limonene can effectively prevent the oxidative damage caused by H2O2 and propose that the main mechanism underlying the inhibitory effects of D-limonene is the inhibition of HLECs apoptosis. In the present study, we used confocal-fluorescence microscopy, flow cytometry analysis, Hoechst staining, H2DCFDA staining, transmission electron microscopy, and immunoblot analysis; the results revealed that slightly higher concentrations of D-limonene (125-1800 μM) reduced the H2O2-induced ROS generation and inhibited the H2O2-induced caspase-3 and caspase-9 activation and decreased the Bcl-2/Bax ratio. Furthermore, it inhibited H2O2-induced p38 MAPK phosphorylation. Thus, we conclude that D-limonene could effectively protect HLECs from H2O2-induced oxidative stress and that its antioxidative effect is significant, thereby increasing the cell survival rate.
由活性氧介导的氧化应激是引发年龄相关性白内障的一个重要因素;D-柠檬烯是一种常见的具有强大抗氧化特性的天然萜烯,广泛存在于多种生物体内。研究表明它具有抗氧化作用;我们发现D-柠檬烯能有效预防过氧化氢引起的氧化损伤,并提出D-柠檬烯抑制作用的主要机制是抑制人晶状体上皮细胞(HLECs)凋亡。在本研究中,我们使用了共聚焦荧光显微镜、流式细胞术分析、Hoechst染色、H2DCFDA染色、透射电子显微镜和免疫印迹分析;结果显示,稍高浓度的D-柠檬烯(125 - 1800μM)可减少过氧化氢诱导的活性氧生成,抑制过氧化氢诱导的半胱天冬酶-3和半胱天冬酶-9激活,并降低Bcl-2/Bax比值。此外,它还抑制过氧化氢诱导的p38丝裂原活化蛋白激酶(p38 MAPK)磷酸化。因此,我们得出结论,D-柠檬烯能有效保护人晶状体上皮细胞免受过氧化氢诱导的氧化应激,其抗氧化作用显著,从而提高细胞存活率。