华西口腔医学杂志 ›› 2018, Vol. 36 ›› Issue (2): 123-127.doi: 10.7518/hxkq.2018.02.002

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氢气对脂多糖致人牙周膜细胞氧化应激损伤的保护作用

周敏(), 王佐林()   

  1. 同济大学口腔医学院·同济大学附属口腔医院牙周科,上海牙组织修复与再生工程技术中心,上海 200072
  • 收稿日期:2017-10-13 修回日期:2018-01-30 出版日期:2018-04-10 发布日期:2018-04-10
  • 作者简介:

    周敏,主治医师,博士,E-mail:kqzhoumin@tongji.edu.cn

  • 基金资助:
    国家自然科学基金(81271110);国家科技支撑计划(20-14BAI04B07);中央高校基本科研业务费专项资金(20152957);上海市卫生计生委青年科研项目(20124y055);同济大学青年优秀人才培养行动计划(1504219041)

Protective effects of hydrogen-rich medium on lipopolysaccharides-induced injury in human periodontal ligament cells

Min Zhou(), Zuolin. Wang()   

  1. Dept. of Periodontology, School and Hospital of Stomatology, Tongji University, Shanghai Engi-neering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China
  • Received:2017-10-13 Revised:2018-01-30 Online:2018-04-10 Published:2018-04-10
  • Supported by:
    Supported by: The National Natural Science Foundation of China (81271110);Chinese National Science Technology Support Program (2014BAI04B07);The Fundamental Research Funds for the Central Universities (20152957);The Shanghai Health Bureau Youth Research Project (20124y055);Program for Young Excellent Talents in Tongji University (1504219041). Correspondence: Wang Zuolin, E-mail: zuolin@tongji.edu.cn.

摘要:

目的 研究培养基中加入氢气能否对人牙周膜细胞(hPDLCs)在脂多糖(LPS)刺激下起到保护作用,降低氧化应激损伤,减少细胞凋亡。方法 用1 μg·mL-1 LPS刺激hPDLCs,分别用普通和富氢培养基培养,检测2组细胞的增殖,凋亡和细胞上清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)水平。结果 在LPS刺激下,富氢培养基组细胞增殖活性显著高于普通培养基组,凋亡率降低(P<0.05)。2组间LDH释放量差异无统计学意义。6 h和12 h富氢培养基组细胞上清中CAT活性较普通培养基组显著升高(分别为P<0.05,P<0.01);而2组的SOD水平在各个时间点均无统计学差异。6 h富氢培养基组细胞上清中MDA水平较普通培养基组显著降低(P<0.05)。结论 氢气可有效改善LPS导致的hPDLCs增殖活性降低及凋亡,并可显著降低氧化应激损伤。

关键词: 富氢培养基, 脂多糖, 牙周膜细胞, 氧化损伤

Abstract:

Objective In this study, lipopolysaccharides (LPS) was used to damage human periodontal ligament cells (hPDLCs) and consequently investigate the protective effects of hydrogen on reducing oxidative stress and cell apoptosis rate. Methods hPDLCs were isolated, and then cultured with normal medium+1 μg·mL-1 LPS or with hydrogen-rich medium+1 μg·mL-1 LPS. Cell proliferation activity was assessed using a cell counting kit-8 (CCK-8), and lactic dehydrogenase (LDH) release was also detected. The activities of superoxide dismutase (SOD) and catalase (CAT), and the level of malonaldehyde (MDA) in supernatants were also measured. Cell apoptosis was detected by flow cytometry at 24 h after LPS stimulation. Results CCK-8 results showed that hydrogen could significantly improve hPDLCs growth and decrease cell apoptosis under LPS stimulation (P<0.05). However, no significant difference in LDH release was found between the two groups. The CAT levels significantly increased at 6 and 12 h in the hydrogen-rich medium as compared with the normal medium group (P<0.05, P<0.01, respectively). However, SOD levels were not significant different at each time point. At 6 h after LPS stimulation, the MDA levels in the cell supernatant of hydrogen-rich medium group were significantly reduced as compared with those in the normal medium group (P<0.05). Conclusion The hydrogen-rich medium can effectively improve hPDLCs proliferation activity and antioxidant capacity and reduce apoptosis and oxidative stress under LPS stimulation.

Key words: hydrogen-rich medium, lipopolysaccharides, periodontal ligament cells, oxidative damage

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