金雀异黄素对人晶状体上皮细胞氧化损伤的保护作用机制

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摘要目的探讨具有雌激素活性的中药单体金雀异黄素（GEN）对过氧化氢（H2O2）诱导的人晶状体上皮细胞（HLE-B3）氧化损伤的保护作用及其线粒体蛋白质组学规律。方法将HLE-B3分为四组，即正常组、H2O2组、雌二醇（E2）组和GEN组。正常组为正常培养的HLE-B3，H2O2组加入终浓度为300 μmol/L的H2O2，E2组加入终浓度为300 μmol/L的H2O2和10-8 mol/L的E2，GEN组加入终浓度为300 μmol/L的H2O2和10-6 mol/L的GEN。表面增强激光解析离子化飞行时间质谱（SELDI-TOF-MS）联合蛋白质芯片技术检测各组HLE-B3线粒体蛋白质表达谱，分析差异蛋白。结果用H2O2诱导HLE-B3氧化损伤后，出现质荷比为6532和6809两个差异表达的蛋白点，H2O2组的峰值高于正常组，差异有统计学意义（P＜0.05）；E2组质荷比为6532的蛋白点的峰值低于H2O2组，差异有统计学意义（P＜0.05）。GEN组质荷比为6532和6809这两个蛋白点的峰值低于H2O2组，差异有统计学意义（P＜0.05）。结论 GEN能有效防护H2O2诱导的HLE-B3氧化损伤，质荷比为6532的蛋白点可能是GEN防护H2O2诱导的HLE-B3氧化损伤的作用靶点，线粒体RPS29可能是植物雌激素GEN保护上皮细胞氧化损伤的一个线粒体蛋白质，为临床寻求防治老年性白内障的安全有效天然药物提供科学的实验依据。

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关键词 ：金雀异黄素, 线粒体蛋白质组, 晶状体上皮细胞, 老年性白内障

Abstract： Objective To investigate the protective effect of the natural medicinal monomer genistein (GEN) with estrogenic activity against oxidative damage in human lens epithelial cells B3 (HLE-B3) caused by hydrogen peroxide (H2O2) and to pursue the possible mitochondrial proteomic regularity of the protective effect. Methods HLE-B3 were divided into four groups: normal group, H2O2 group, estradiol (E2) group and GEN group. Normal group is normal culture HLE-B3, H2O2 group with final concentration of 300 μmol/L H2O2, E2 group with final concentration of 300 μmol/L H2O2 and 10-8 mol/L E2, the final concentration of 300 μmol/L H2O2 and 10-6 mol/L GEN were added into the GEN group. The mitochondrial protein expression profiles of HLE-B3 in each group were detected by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) combined with protein chip technology, and the differential proteins were analyzed. Results Two differentially expressed protein sites with mass to charge ratios of 6532 and 6809 appeared after oxidative damage of HLE-B3 induced by H2O2. The peak value of H2O2 group was higher than that of normal group, the difference was statistically significant (P<0.05). The peak value of protein point in E2 group with mass/charge ratio of 6532 was lower than that in H2O2 group, the difference was statistically significant (P<0.05). The peak values of protein points 6532 and 6809 in GEN group were lower than those in H2O2 group, the differences were statistically significant (P<0.05). Conclusion GEN could effectively inhibite H2O2 induced oxidative damage in HLE-B3 cells. The protein spot at mass/charge ratio of 6532 might be the target spot of GEN against oxidative damage induced by H2O2. Mitochondrial RPS29 may be a mitochondrial protein that protects epithelial cell from oxidative damage by phytoestrogens GEN. In order to provide scientific basis for pursueing safe and effective natural drugs to prevent and cure senile cataract.

Key words： Genistein Mitochondrial proteomics Lens epithelial cell Senile cataract

基金资助:福建省自然科学基金资助课题（2019J01486）；
福建省卫生健康中青年骨干人才培养项目（2020GGA065）

作者简介: 冯春燕（1979-），女，福建永安人，博士，副主任医师，硕士研究生导师，主要从事眼病的中西医基础与临床研究。

引用本文:

冯春燕；唐宋文；黄秀榕；祁明信；胡艳红. 金雀异黄素对人晶状体上皮细胞氧化损伤的保护作用机制[J]. 中国当代医药, 2022, 29(13): 5-9.
FENG Chunyan； TANG Songwen； HUANG Xiurong； QI Mingxin； HU Yanhong. Mitochondrial proteomic of genistein protection against oxidative damage in human lens epithelial cells. 中国当代医药, 2022, 29(13): 5-9.

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