中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (8): 1228-1233.doi: 10.12307/2022.228

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

石斛多糖保护中波紫外线损伤成纤维细胞的作用机制

李  琴1，2，毛双法1，李  敏2，程基焱2   

  1. 1四川卫生康复职业学院基础医学部，四川省自贡市   633000；2西南医科大学基础医学院，四川省泸州市   646000
  • 收稿日期:2021-04-27 修回日期:2021-04-29 接受日期:2021-06-05 出版日期:2022-03-18 发布日期:2021-11-02
  • 通讯作者: 程基焱，硕士，教授，硕士生导师，西南医科大学基础医学院，四川省泸州市 646000
  • 作者简介:李琴，女，1983年生，四川省达州市人，汉族，2021年西南医科大学毕业，硕士，讲师，主要从事皮肤损伤与修复的研究。
  • 基金资助:
    四川省科学技术厅专项资金计划资助项目(15ZB0160)，项目负责人：程基焱；四川省教育厅资金计划资助项目，2014-LY-05，项目负责人：程基焱

Protective effect and mechanism of dendrobium on fibroblasts damaged by ultraviolet B

Li Qin1, 2, Mao Shuangfa1, Li Min2, Cheng Jiyan2   

  1. 1Department of Basic Medicine, Sichuan Health Rehabilitation Vocational College, Zigong 633000, Sichuan Province, China; 2College of Basic Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2021-04-27 Revised:2021-04-29 Accepted:2021-06-05 Online:2022-03-18 Published:2021-11-02
  • Contact: Cheng Jiyan, Master, Professor, Master’s supervisor, College of Basic Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Li Qin, Master, Lecturer, Department of Basic Medicine, Sichuan Health Rehabilitation Vocational College, Zigong 633000, Sichuan Province, China; College of Basic Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    the Science and Technology Department of Sichuan Province, No. 15ZB0160 (to CJY); a grant from the Education Department of Sichuan Province, No. 2014-LY-05 (to CJY)

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摘要：

文题释义：
石斛多糖(dendrobium)：中国传统中药石斛的主要单体活性成分，据报道可提高抗氧化酶活性、清除自由基、抑制脂质过氧化的活性，对延缓衰老有一定的作用。石斛多糖广泛应用于呼吸系统、神经系统、免疫系统、医学美容等方面研究。
超氧化物歧化酶(superoxide dismutase，SOD)：可以催化活性氧的超氧阴离子自由基歧化反应，在部分细胞内的氧化与抗氧化平衡调节、防止光损伤、光老化损伤起关键作用。
背景：中波紫外线导致活性氧升高，是光损伤和光老化最重要的物理因素。石斛所含有的石斛多糖可提高抗氧化酶活性、清除自由基、抑制脂质过氧化的活性，对稳定生物膜及延缓衰老有一定的作用。
目的：探讨石斛多糖对中波紫外线诱导体外培养人成纤维细胞光损伤的保护作用及可能机制。
方法：体外培养人成纤维细胞，将其随机分为4组：对照组细胞正常培养；模型组细胞进行紫外线一次性照射，照射剂量为20 mJ/cm2；石斛组培养基中加入质量浓度30 mg/L石斛多糖干扰培养；实验组细胞用照射剂量为20 mJ/cm2紫外线照射后，加入30 mg/L石斛多糖干扰培养。各组细胞分别处理后培养48 h。用qRT-PCR、Western blot、免疫组化检测Nrf2、超氧化物歧化酶基因和蛋白表达，用荧光检测细胞活性氧水平，比色法测定丙二醛含量。
结果与结论：①与对照组比较，模型组、石斛组、实验组Nrf2和超氧化物歧化酶的mRNA及蛋白表达明显上调(均P < 0.05)，实验组显著高于模型组和石斛组(P < 0.05)；②模型组活性氧明显升高(P < 0.05)，石斛组活性氧降低(P < 0.05)，实验组活性氧明显低于模型组(P < 0.05)；③与对照组比较，模型组、实验组丙二醛明显升高(P < 0.05)，石斛组明显降低(P < 0.05)，实验组明显低于模型组(P < 0.05)；④结果说明，石斛多糖对中波紫外线导致体外培养人成纤维细胞光老化和光损伤具有一定的保护作用，其主要作用机制是通过Keap1-Nrf2-ARE信号通路调控超氧化物歧化酶的产生，清除活性氧，减少中波紫外线对细胞的损伤。

https://orcid.org/0000-0003-2988-6848 (李琴) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 光损伤, 光损伤, 成纤维细胞, 石斛多糖, 超氧化物歧化酶, 活性氧, Nrf2, 丙二醛

Abstract: BACKGROUND: Ultraviolet B increases reactive oxygen species, which is the most important physical factor of light damage and photoaging. Dendrobium polysaccharides contained in dendrobium can improve the activity of antioxidant enzymes, scavenge free radicals, and inhibit lipid peroxidation, which have certain effects on stabilizing biofilms and delaying aging.
OBJECTIVE: To investigate the preventive effect and mechanism of dendrobium on the photoaging fibroblasts of human induced by ultraviolet B in vitro. 
METHODS: Human fibroblasts cultured in vitro were randomized into four groups: a control group with normal culture, a model group irradiated by one-time ultraviolet B at a dose of 20 mJ/cm2, a dendrobium group interfered with 30 mg/L dendrobium polysaccharides, and an experimental group interfered with 30 mg/L dendrobium polysaccharides following 20 mJ/cm2 ultraviolet B irradiation. Cells in each group were then cultured for another 48 hours. Expression of Nrf2 and superoxide dismutase at gene and protein levels was detected using qRT-PCR, western blot and immunohistochemistry. Cellular reactive oxygen levels were detected using fluorescence detection, and malondialdehyde content was detected using colorimetric determination. 
RESULTS AND CONCLUSION: The mRNA and protein levels of Nrf2 and superoxide dismutase were significantly up-regulated in the model, dendrobium and experimental groups compared with the control group (P < 0.05). The mRNA and protein levels of Nrf2 and superoxide dismutase were significantly higher in the experimental group than the model and dendrobium groups (P < 0.05). Compared with the control group, the reactive oxygen level was significantly increased in the model group (P < 0.05), and decreased in the dendrobium group (P < 0.05). The reactive oxygen level in the experimental group was significantly lower than that in the model group (P < 0.05). Compared with the control group, the malondialdehyde content was significantly increased in the model and experimental groups (P < 0.05), and significantly decreased in the dendrobium group (P < 0.05). The malondialdehyde content in the experimental group was significantly lower than that in the model group (P < 0.05). To conclude, dendrobium has certain protective effects on the photoaging of human fibroblasts induced by ultraviolet B. The main mechanism of dendrobium may be to regulate the production of superoxide dismutase, and eliminate reactive oxygen, reduce cell damage caused by ultraviolet B through Keap1-Nrf2-ARE signaling pathway.


Key words: photoagin, fibroblasts, dendrobium, dendrobium polysaccharides, superoxide dismutase, reactive oxygen, Nrf2, malondialdehyde

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