中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (12): 856-859.

• 论著 • 上一篇    下一篇

黄芪甲苷对HaCaT细胞的光保护效能及其机制

杨子良1,骆丹2,钱齐宏3,杜纳4,余秀琴4,王淼淼5,闵玮4   

  1. 1. 苏州大学医学院附属第一人民医院皮肤科
    2. 南京市南京医科大学附属第一医院皮肤科
    3. 苏州大学附属第一医院皮肤科
    4. 苏州大学附属第一医院
    5. 中国医学科学院皮肤病研究所
  • 收稿日期:2014-01-13 修回日期:2014-09-10 出版日期:2014-12-15 发布日期:2019-06-14
  • 通讯作者: 闵玮 E-mail:ztmw_2001@126.com
  • 基金资助:
    江苏省自然科学基金;江苏省卫生厅中医药科技项目;苏州市科教兴卫项目

Protective effect of astragaloside Ⅳ against ultraviolet B-induced photodamage to human HaCaT keratinocytes and its mechanisms

  • Received:2014-01-13 Revised:2014-09-10 Online:2014-12-15 Published:2019-06-14
  • Contact: Wei Min E-mail:ztmw_2001@126.com
  • Supported by:
    Natural Science Foundation of Jiangsu Province of China

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摘要： 目的 观察黄芪甲苷对于中波紫外线（UVB）损伤的人表皮细胞的保护作用，探讨其相关机制。方法 将培养的永生化人皮肤角质形成细胞（HaCaT细胞）分为对照组、UVB组、黄芪甲苷组和UVB + 黄芪甲苷组，其中UVB组和UVB + 黄芪甲苷组细胞接受50 mJ/cm2 UVB照射，加药组加入不同浓度的黄芪甲苷（10、20、50、100、200 mg/L）进行干预，24 h后CCK8法检测细胞活性。根据CCK8法检测结果选择最佳药物浓度（20 mg/L）进行后继实验。照光后继续培养24 h，流式细胞仪检测细胞内活性氧（ROS）水平，Western印迹法检测HaCaT细胞中p53、p38、基质金属蛋白酶9（MMP-9）和高迁移率族蛋白A1（HMGA-1）的表达。 结果 与对照组相比，10 mg/L和20 mg/L黄芪甲苷组对HaCaT细胞的增殖活性无明显影响（F = 1.32，P > 0.05），50、100和200 mg/L黄芪甲苷对细胞增殖活性有一定抑制作用（F = 20.20，P < 0.05）；UVB组与对照组比较，HaCaT细胞增殖活性显著下降（F = 99.00，P < 0.01）。与UVB组相比，UVB + 黄芪甲苷（10 ~ 200 mg/L）组HaCaT细胞增殖活性不同程度升高（F = 19.08，P < 0.01），其中UVB + 20 mg/L黄芪甲苷组升高程度最高。进一步实验表明，与UVB组相比，UVB + 20 mg/L黄芪甲苷组ROS产生受到明显抑制（t = 21.12，P < 0.01）。Western印迹结果表明，与对照组比较，UVB组p53、p38、MMP-9和HMGA-1蛋白的表达升高（均P < 0.01），而UVB + 20 mg/L黄芪甲苷组细胞内p53、p38、MMP-9和HMGA-1蛋白的表达水平较UVB组显著降低（P < 0.01）。 结论 黄芪甲苷可有效抑制UVB引起的表皮细胞光损伤。

关键词: 黄芪, 紫外线, 角蛋白细胞, 活性氧

Abstract: Yang Ziliang*, Luo Dan, Qian Qihong, Du Na, Yu Xiuqin, Wang Miaomiao, Min Wei. *Department of Dermatology, First Affiliated Hospital of Soochow University, Suzhou 215006, China Corresponding author: Min Wei, Email: minwei@suda.edu.cn 【Abstract】 Objective To evaluate the protective effect of astragaloside Ⅳ against ultraviolet B （UVB）-induced photodamage to human HaCaT keratinocytes, and to investigate its mechanisms. Methods Cultured immortalized human HaCaT keratinocytes were divided into four groups: blank control group receiving untreated, UVB group irradiated with 50 mJ/cm2 UVB, astragaloside Ⅳ group treated with astragaloside Ⅳ, UVB + astragaloside Ⅳ group treated with astragaloside Ⅳ for 24 hours before and after 50 mJ/cm2 of UVB radiation. The concentration of astragaloside Ⅳ ranged from 10 to 200 mg/L in cell proliferation assay, and according to the results of proliferation assay, 20 mg/L was determined as the optimal concentration in the other assays. At 24 hours after UVB radiation, cell counting kit-8 （CCK8） assay was performed to evaluate cellular proliferative activity, flow cytometry to determine intracellular reactive oxygen species （ROS） levels, and Western blot to measure the expression levels of p53, p38, matrix metalloproteinase-9 （MMP-9） and high mobility group Al （HMGA-1） protein in HaCaT cells. Results Compared with the control group, astragaloside Ⅳ at 10 and 20 mg/L had no inhibitory effect （F = 1.32, P > 0.05）, while astragaloside Ⅳ at 50, 100 and 200 mg/L showed significantly inhibitory effect （F = 20.20, P < 0.05）, on the proliferation of HaCaT cells. In addition, cellular proliferative activity in the UVB group was significantly lower than that in the control group （F = 99.00, P < 0.01）. Compared with the UVB group, cellular proliferative activity increased to different degrees in HaCaT cells treated with both UVB and astragaloside Ⅳ of 10－200 mg/L （F = 19.08, P < 0.01）, with the strongest increase observed in those treated with UVB and astragaloside Ⅳ of 20 mg/L. Further experiments revealed reduced intracellular ROS levels in the UVB + astragaloside Ⅳ （20 mg/L） group compared with the UVB group （t = 21.12, P < 0.01）. Western blot assay showed that the expression levels of p53, p38, MMP-9 and HMGA-1 protein were significantly higher in the UVB group than in the control group （all P < 0.01）, but significantly lower in the UVB + astragaloside Ⅳ （20 mg/L） group than in the UVB group （all P < 0.01）. Conclusion Astragaloside Ⅳ can effectively protect keratinocytes from UVB-induced photodamage.

Key words: Astragalus membranaceus, Ultraviolet rays, Keratinocytes, Reactive oxygen species