国际口腔医学杂志

国际口腔医学杂志 ›› 2022, Vol. 49 ›› Issue (5): 521-528.doi: 10.7518/gjkq.2022080

• 论著 • 上一篇    下一篇

黄芩苷抑制脂多糖促巨噬细胞氧化应激损伤作用的研究

黄伟琨(),徐秋艳,周婷()   

  1. 贵州省人民医院口腔科 贵阳 550002
  • 收稿日期:2022-01-10 修回日期:2022-05-18 出版日期:2022-09-01 发布日期:2022-09-16
  • 通讯作者: 周婷
  • 作者简介:黄伟琨，副主任医师，硕士，Email：hwk8467@163.com
  • 基金资助:
    国家自然科学基金(81760198);贵州省中医药、民族医药科学技术研究专项课题(QZYY-2019-011);贵州省人民医院青年基金(GZSYQN[2018]08);贵州省卫生健康委员会科学技术基金(gzwjkj2018-1-054);贵州省人民医院国家自然科学基金培育基金(QKHPTRC[2018]5764-06)

Role of baicalin and mechanisms through which baicalin attenuates oxidative stress injury induced by lipopolysaccharide on macrophages

Huang Weikun(),Xu Qiuyan,Zhou Ting.()   

  1. Dept. of Stomatology, Guizhou Provincial People ’ s Hospital, Guiyang 550002, China
  • Received:2022-01-10 Revised:2022-05-18 Online:2022-09-01 Published:2022-09-16
  • Contact: Ting. Zhou
  • Supported by:
    National Natural Science Foundation of China(81760198);Science and Technology Foundation of the Traditional Chinese Medicine of Guizhou Province(QZYY-2019-011);Youth Fund of Guizhou Provincial People’s Hospital(GZSYQN[2018]08);Science and Technology Foundation of the Health Commission of Guizhou Province(gzwjkj2018-1-054);Cultivation Fund of National Natural Science Foundation of Guizhou Provincial People’s Hospital(QKHPTRC[2018]5764-06)

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摘要：

目的 研究黄芩苷在牙龈卟啉单胞菌（P. gingivalis）的致病因子脂多糖（LPS）促巨噬细胞氧化应激反应中的作用及分子机制。 方法 LPS刺激巨噬细胞的同时，采用不同浓度（5、10 μmol·L-1）的黄芩苷进行干预，分别采用细胞活性检测试剂盒（CCK8）、乳酸脱氢酶（LDH）试剂盒、2,7-二氢二氯荧光黄（DCFA-DA）探针、丙二醛（MDA）、超氧化物歧化酶（SOD）试剂盒、流式细胞仪，检测P. gingivalis-LPS对细胞所造成的损伤、细胞内活性氧（ROS）含量、MDA和SOD活性、细胞凋亡；随后，采用蛋白质印迹法（WB）检测核因子E2相关因子2（Nrf2）的总蛋白、胞质蛋白及胞核蛋白的表达情况。 结果 与对照组相比，P. gingivalis-LPS可以导致细胞活力下降（P<0.000 1）、LDH含量上升（P<0.000 1）、ROS和MDA含量上调（P<0.000 1）、SOD活性下降（P<0.000 1）以及细胞凋亡率上升（P<0.000 1）。黄芩苷的干预能够减轻P. gingivalis-LPS对细胞所造成的上述损伤（P<0.000 1）；10 μmol·L-1黄芩苷的干预能够显著上调胞核Nrf2的表达（P<0.01）。 结论 黄芩苷通过促进巨噬细胞Nrf2的核转位，减轻了P. gingivalis-LPS所诱发的氧化应激损伤。

关键词: 黄芩苷, 牙周炎, 氧化应激, 巨噬细胞

Abstract:

Objective We aimed to explore the role and mechanisms of baicalin in oxidative injury promoted by Porphyromonas gingivalis (P. gingivalis) lipopolysaccharides (LPSs) on macrophages. Methods Macrophages derived from human monocytic-leukemia cells (THP-1) were stimulated with LPSs with or without baicalin (5 and 10 μmol·L-1), and then cell injury, reactive oxygen species (ROS) level, malondialdehyde (MDA), superoxide dismutase (SOD) activities, and apoptosis were measured through cell counting kit-8 (CCK8), lactate dehydrogenase (LDH), 2,7-dichlorodihydrofluorescein diacetate, MDA, and SOD assays; enzyme-linked immunosorbent assay; and flow cytometry. The expression level of nuclear factor erythroid 2-related factor 2 (Nrf2) was determined by Wes-tern Blot (WB). Results LPS inhibited cell proliferation (P<0.000 1), induced LDH leakage (P<0.000 1), increased ROS level and MDA activity (P<0.000 1), decreased SOD activity (P<0.000 1), and increased apoptotic rate (P<0.000 1). Baicalin co-treatment attenuated damage induced by LPSs (P<0.000 1). Meanwhile, the cytoplasmic protein level of Nrf2 was up-regulated by 10 μmol·L-1 baicalin co-treatment (P<0.01). Conclusion Baicalin attenuated LPS-promoted oxidative injury on macrophages by up-regulating Nrf2.

Key words: baicalin, periodontitis, oxidative injury, macrophage

中图分类号: 

  • Q 26

图 1

黄芩苷对LPS致巨噬细胞损伤的影响*P<0.05，***P<0.001，****P<0.000 1。"

图 2

黄芩苷对LPS致巨噬细胞氧化应激反应的作用**P<0.01，***P<0.001，****P<0.000 1，ns为无统计学意义。"

图 3

不同浓度黄芩苷对LPS致巨噬细胞凋亡的影响**P<0.01，****P<0.000 1。"

图 4

黄芩苷抑制LPS致巨噬细胞氧化应激损伤及细胞凋亡的分子机制*P<0.05，**P<0.01，ns为无统计学意义。"

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