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日用化学工业（中英文）

日用化学工业（中英文） ›› 2022, Vol. 52 ›› Issue (11): 1195-1201.doi: 10.3969/j.issn.1001-1803.2022.11.007

• 开发与应用 • 上一篇    下一篇

绿原酸通过SIRT6改善UVB诱导的HaCaT炎症反应

武思敏1,2,满佳旭1,2,袁梅1,2,高梓琪1,2,王冬雪1,2,张冬英1,3,*()   

  1. 1.云南农业大学 普洱茶学教育部重点实验室，云南 昆明 650201
    2.云南农业大学 食品科学与技术学院，云南 昆明 650201
    3.云南农业大学 理学院，云南 昆明 650201
  • 收稿日期:2021-12-01 修回日期:2022-11-01 出版日期:2022-11-22 发布日期:2022-11-22
  • 通讯作者: 张冬英
  • 基金资助:
    国家自然科学基金资助项目(31760226);云南省重大科技项目(2018IA060)

Chlorogenic acid ameliorates UVB-induced HaCaT inflammation through SIRT6

Wu Simin1,2,Man Jiaxu1,2,Yuan Mei1,2,Gao Ziqi1,2,Wang Dongxue1,2,Zhang Dongying1,3,*()   

  1. 1. Key Laboratory of Puer Tea, Ministry of Education, Yunnan Agricultural University, Kunming, Yunnan 650201, China
    2. College of Food Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, China
    3. College of Science, Yunnan Agricultural University, Kunming, Yunnan 650201, China
  • Received:2021-12-01 Revised:2022-11-01 Online:2022-11-22 Published:2022-11-22
  • Contact: Dongying Zhang

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摘要：

研究绿原酸（CGA）对UVB诱导的HaCaT细胞炎症反应的调节作用及机制。用CGA干预UVB处理的HaCaT细胞，结晶紫染色法检测CGA对UVB损伤HaCaT细胞的保护作用。通过酶联免疫吸附法检测细胞内TNF-α含量的变化，ABTS、DPPH、FRAP法检测CGA在细胞内外的抗氧化能力，丙二醛试剂盒法检测细胞中MDA含量，CM-H2DCFDA染色法检测细胞内总活性氧的变化，Western Blot法检测SIRT6、TNF-α、P-AKT、P-AMPK、AMPK、β-Tubulin蛋白的相对表达量。结晶紫染色表明100和150 μmol/L CGA对于21.6 mJ/cm2 UVB处理的HaCaT细胞具有保护作用。酶联免疫吸附法证明了CGA处理能够显著降低TNF-α的表达（P<0.01）。CGA在细胞内外均具备良好的自由基清除能力，并且能够有效降低UVB诱导的MDA和ROS的产生。CGA可以显著抑制由UVB引起的SIRT6表达上调，从而降低AKT的磷酸化水平，活化AMPK进而降低TNF-α的表达，缓解细胞的炎症反应。CGA可以抑制SIRT6的表达进而降低UVB诱导的HaCaT的炎症反应，并且降低UVB造成的细胞的氧化应激，具有很强的抗皮肤光老化活性。

关键词: 绿原酸, 组蛋白脱乙酰化蛋白-6（SIRT6）, 肿瘤坏死因子-α（TNF-α）, 炎症, 皮肤光老化

Abstract:

The aim of the study was to investigate the regulatory effect and mechanism of chlorogenic acid （CGA） on UVB-induced inflammatory response in HaCaT cells. HaCaT cells exposed to UVB were treated with CGA, and the protective effect of CGA on HaCaT cells damaged by UVB was detected by crystal violet staining. Changes in intracellular TNF-α content were detected by enzyme-linked immunosorbent assay, the antioxidant capacity of CGA in and out of the cells was detected by ABTS, DPPH, and FRAP assay, MDA content in the cells was detected by malondialdehyde kit, changes in total intracellular reactive oxygen species were detected by CM-H2DCFDA staining, and the relative expression of SIRT6, TNF-α, P-AKT, P-AMPK, AMPK, β-Tubulin proteins were detected by Western blot assay. Crystal violet staining and microscopy indicate that 100 and 150 μmol/L CGA are protective for 21.6 mJ/cm2 UVB-treated HaCaT cells. Enzyme-linked immunosorbent assay results indicate that CGA treatment significantly reduces TNF-α expression （P<0.01）. CGA has good free radical scavenging ability both inside and outside cells, and is able to effectively reduce the UVB induced production of MDA and ROS. CGA can significantly inhibit the upregulation of SIRT6 expression caused by UVB, thereby reducing the phosphorylation level of AKT, activating AMPK and then reducing the expression of TNF-α, and alleviating the inflammatory response of the cells. CGA can inhibit SIRT6 to reduce the inflammatory response of UVB-induced HaCaT cells and reduce UVB-inflicted cellular oxidative stress with strong anti-cutaneous photoaging activity.

Key words: chlorogenic acid, SIRT6, TNF-α, inflammation, skin photoaging

中图分类号: 

  • TQ658