| 摘要: |
| 目的 探讨富氢水对甲状腺相关眼病（TAO）的抗氧化应激作用。方法 取TAO患者眼眶脂肪结缔组织的眼眶成纤维细胞进行体外培养，用不同浓度的过氧化氢（H2O2）处理细胞18 h后，用CCK-8法检测细胞增殖能力以确定合适的H2O2浓度。将细胞为4组：空白组（正常培养）、H2O2组（H2O2处理18 h）、富氢水+H2O2组（富氢水处理72 h的同时用H2O2处理18 h）、地塞米松+H2O2组（1 μmol/L地塞米松处理72 h的同时用H2O2处理18 h）。用流式细胞术检测各组细胞活性氧（ROS）荧光强度，用ELISA检测细胞培养液中丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）的含量。结果 TAO患者眼眶成纤维细胞体外培养成功。H2O2浓度越高对TAO患者眼眶成纤维细胞增殖能力的抑制效果越明显，本实验最终选取的H2O2浓度为100 μmol/L。培养TAO患者眼眶成纤维细胞72 h后，空白组、H2O2组、富氢水+H2O2组、地塞米松+H2O2组细胞培养液中MDA、SOD、GSH-Px的含量和细胞ROS荧光强度分别为（1.63±0.29）、（5.06±0.24）、（3.94±0.29）、（2.34±0.24）nmol/mL，（10.51±0.32）、（2.41±0.23）、（5.58±0.29）、（7.98±0.15）U/mL，（107.79±1.06）、（21.07±0.92）、（49.19±6.75）、（76.33±4.70）U/mL和18 275.82±521.05、92 524.81±2 097.01、54 460.87±572.64、35 961.37±540.61，统计分析发现富氢水和地塞米松均可抑制H2O2处理后眼眶成纤维细胞的氧化应激（P均<0.01），且地塞米松的抑制效果较富氢水更明显（P均<0.01）。结论 氢分子可能通过抗氧化应激对TAO发挥治疗作用。 |
| 关键词: 甲状腺相关眼病 眼眶成纤维细胞 富氢水 氧化性应激 地塞米松 |
| DOI：10.16781/j.0258-879x.2018.05.0547 |
| 投稿时间：2018-03-09修订日期：2018-04-25 |
| 基金项目:国自然青年科学基金：免疫抑制剂冲击治疗影响甲状腺相关眼病患者泪液蛋白质组学的研究。课题编码；81500758资助 |
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| Effect of hydrogen-rich water on oxidative stress of in vitro orbital fibroblasts of patients with thyroid associated ophthalmopathy |
| CHEN Xin-xin,CAI Ji-ping,CHENG Jin-wei,ZHOU Xiao-qing,SHEN Ya,LI Jian,WEI Rui-li* |
(Department of Ophthalmology, Changzheng Hospital, Navy Medical University (Second Military Medical University), Shanghai 200003, China
*Corresponding author) |
| Abstract: |
| Objective To explore the anti-oxidation stress effect of hydrogen-rich water (HW) on thyroid associated ophthalmopathy (TAO). Methods The orbital fibroblasts were derived from orbital adipose connective tissue of TAO patients and cultured in vitro. The cells were treated with different concentrations of hydrogen peroxide (H2O2) for 18 h, and the proliferation ability was detected by CCK-8 method to determine the appropriate H2O2 concentration. The cells were divided into four groups:blank group (normal culture), H2O2 group (treating cells with H2O2 for 18 h), HW+H2O2 group (culturing cells using culture media containing HW for 72 h in combination with H2O2 for 18 h), dexamethasone+ H2O2 group (treating cells using dexamethasone 1 μmol/L for 72 h in combination with H2O2 for 18 h). After culturing for 72 h in each group, the fluorescence intensity of reactive oxygen species (ROS) was measured by flow cytometry, and the contents of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by ELISA. Results We successfully cultured orbital fibroblasts derived from orbital adipose connective tissues of TAO patients. The higher concentration of H2O2, the greater inhibition effect on the proliferation ability of the orbital fibroblasts from TAO patients, and we finally chose 100 μmol/L H2O2. After 72 h of cell culture, the contents of MDA, SOD, and GSHPx and the fluorescence intensity of ROS were (1.63±0.29), (5.06±0.24), (3.94±0.29), and (2.34±0.24) nmol/mL, (10.51±0.32), (2.41±0.23), (5.58±0.29), and (7.98±0.15) U/mL, (107.79±1.06), (21.07±0.92), (49.19±6.75), and (76.33±4.70) U/mL and 18 275.82±521.05, 92 524.81±2 097.01, 54 460.87±572.64, and 35 961.37±540.61 in the blank, H2O2, HW+ H2O2 and dexamethasone+H2O2 groups, respectively. Statistic analysis results showed that both HW and dexamethasone could significantly inhibit oxidative stress induced by H2O2 in orbital fibroblasts (all P<0.01), and the inhibitory effects of dexamethasone were significantly more obvious than those of HW (all P<0.01). Conclusion HW may be a treatment option for TAO through anti-oxidant stress. |
| Key words: thyroid associated ophthalmopathy orbital fibroblasts hydrogen-rich water oxidative stress dexamethasone |
