| 南珠水解液对H2O2诱导的人晶状体上皮细胞氧化损伤的保护作用 |
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| 引用本文： | 刘鹏,岑妍慧,林江,兰太进.南珠水解液对H2O2诱导的人晶状体上皮细胞氧化损伤的保护作用[J].眼科新进展,2020,0(7):612-616. |
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| 作者姓名： | 刘鹏 岑妍慧 林江 兰太进 |
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| 作者单位： | 530200 广西壮族自治区南宁市，广西中医药大学基础医学院 |
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| 摘 要： | 目的 探讨南珠水解液对过氧化氢（H2O2）诱导的人晶状体上皮细胞（HLEC）氧化应激损伤的保护作用。方法 培养HLEC细胞株HLEB3，在培养液中加入不同浓度南珠水解液，培养24 h和48 h分别检测细胞增殖水平，并据此选择后续实验中南珠水解液浓度。培养HLE-B3细胞，分成3组:正常对照组；氧化损伤组，加入H2O2至终浓度为300 μmol·L-1；南珠水解液处理组，加入H2O2至终浓度为300 μmol·L-1，加入南珠水解液至终浓度为200 mg·L-1。3组细胞使用流式细胞术检测细胞凋亡率，利用电镜观察细胞形态差异；利用生化检测试剂盒检测3组细胞中谷胱甘肽过氧化物酶（GSH-Px）和超氧化物岐化酶（SOD）活性以及丙二醛（MDA）含量。结果 3组细胞增殖法检测结果显示，南珠水解液处理HLE-B3细胞24 h后，对其活性无抑制作用；50 mg·L-1、100 mg·L-1、200 mg·L-1南珠水解液处理后细胞存活率分别为（128.68±12.35）%、（158.43±11.71）%和（164.16±24.46）%，48 h后能显著提高其增殖水平，与正常对照组［（100.00±9.78）%］比较，差异有统计学意义（P<0.05）。流式细胞仪检测结果显示，正常对照组细胞凋亡率为（4.527±0.321）%；氧化损伤组为（11.460±0.633）%；南珠水解液处理组细胞凋亡率降至（8.877±0.193）%，与氧化损伤组相比差异具有统计学意义（P<0.05）。电镜下观察结果显示，H2O2氧化损伤可以诱导HLE-B3细胞形态改变，损伤程度最高；南珠水解液处理后，细胞形态轻微改变。氧化损伤组细胞内GSH-Px、SOD含量均降低，MDA含量升高，与正常对照组相比差异均具有统计学意义（均为P<0.05)；南珠水解液处理组GSH-Px、SOD含量均升高，MDA含量下降，与氧化损伤组相比差异均具有统计学意义（均为P<0.05)。结论 南珠水解液能够促进HLEC生长，抑制细胞凋亡，能够提升HLEC中GSH-Px、SOD活性，降低MDA含量，对H2O2诱导的HLEC氧化损伤具有保护作用。
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| 关 键 词： | 南珠水解液 人晶状体上皮细胞 氧化损伤 保护作用 |
Protective effect of Nanzhu hydrolysate against H2O2-induced oxidative damage in human lens epithelial cells |
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| LIU Peng,CEN Yanhui,LIN Jiang,LAN Taijin.Protective effect of Nanzhu hydrolysate against H2O2-induced oxidative damage in human lens epithelial cells[J].Recent Advances in Ophthalmology,2020,0(7):612-616. |
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| Authors: | LIU Peng CEN Yanhui LIN Jiang LAN Taijin |
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| Affiliation: | Basic Medical College of Guangxi University of Traditional Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China |
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| Abstract: | Objective To investigate the protective effect of Nanzhu hydrolysate against oxidative stress in human lens epithelial cells (HLEC) induced by hydrogen peroxide (H2O2). Methods The HLE-B3 cells were cultured, and different concentrations of Nanzhu hydrolysate was added into the culture medium. Cell proliferation levels were detected after 24 h and 48 h, and the concentration of Nanzhu hydrolysate in subsequent experiments was selected accordingly. The HLE-B3 cells were cultured and divided into three groups: control group, oxidative damage group and Nanzhu hydrolysate-treatment group. In the oxidative damage group, the final concentration of H2O2 was 300 μmol·L-1. In the Nanzhu hydrolysate-treatment group, the final concentration of H2O2 was 300 μmol·L-1, and the final concentration of Nanzhu hydrolysate was 200 mg·L-1. Apoptosis rate of the HLE-B3 cells was detected by flow cytometry, while the cell morphological differences of the three groups were observed by electron microscope. The enzyme activity and content of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) andmalondialdehyde (MDA) in cells were detected by biochemical detection kit. Results CCK8 methods showed that the activity of HLE-B3 cells was not inhibited in the Nanzhu hydrolysate-treatment group after 24 h. After 50 mg·L-1, 100 mg·L-1, 200 mg·L-1 Nanzhu hydrolysate treatment for 48 h, the proliferation level was significantly improved, and the survival rates were (128.68±12.35) %, (158.43±11.71) % and (164.16±24.46) %, respectively, which were significantly different from the control group ［(100.00±9.78) %］. The results of flow cytometry showed that the apoptosis rate was (4.527±0.321)%, (11.460±0.633) %, (8.877±0.193) % in the control group, oxidative damage group and Nanzhu hydrolysate-treatment group, respectively, and Nanzhu hydrolysate-treatment group was significantly different from the oxidative damage group. Ultramicroscopic results showed that H2O2 oxidative damage could induce the morphological changes of HLE-B3 cells, with the highest degree of damage after H2O2, slightly morphological changes after Nanzhu hydrolysate treatment. The content of GSH-Px and SOD in the cells of the oxidative damage group decreased, and the content of MDA increased, which was significantly different from those of the control group. The content of GSH-Px and SOD in the Nanzhu hydrolysate-treatment group increased, while the content of MDA decreased, which was significantly different from those in the oxidative damage group. Conclusion Nanzhu hydrolysate-treatment can promote cell growth, inhibit cell apoptosis, enhance GSH-Px and SOD activity in human lens epithelial cells, reduce MDA content, and protect against oxidative damage induced by H2O2. |
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| Keywords: | Nanzhu hydrolysate human lens epithelial cells oxidative damage protective effect |
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