生物学杂志

生物学杂志 ›› 2024, Vol. 41 ›› Issue (2): 39-.doi: 10.3969/j.issn.2095-1736.2024.02.039

• 研究报告 • 上一篇    下一篇

川藏香茶菜庚素的体外抗炎作用与分子机制研究

杨东娟， 查广才， 邹湘辉， 吴丹彤， 伍清婷   

  1. 韩山师范学院 生命科学与食品工程学院， 潮州 521041
  • 出版日期:2024-04-18 发布日期:2024-04-17
  • 作者简介:杨东娟，硕士研究生，高级实验师，主要研究方向为细胞生物学及分子药理学，E-mail:dongjuan_1235@126.com
  • 基金资助:
    韩山师范学院一般项目(LY202601)； 广东省粤东药食资源功能物质与治未病研究重点实验室(2021B1212040015)； 广东省普通高校重点科研项目(2022ZDZX4030)

Anti-inflammatory effect and mechanism of Pseurata H on inflammation model of LPS-induced RAW264.7 cells

YANG Dongjuan, ZHA Guangcai, ZOU Xianghui, WU Dantong, WU Qingting   

  1. School of Life Sciences and Food Engineering, Hanshan Normal University, Chaozhou 521041, China
  • Online:2024-04-18 Published:2024-04-17

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摘要： 为探究川藏香茶菜庚素(Pseurata H)对巨噬细胞炎症模型的抗炎作用及分子机制，利用LPS刺激RAW264.7细胞建立体外炎症模型，并分别以浓度为10、20和40 μmol/L的Pseurata H干预该细胞模型。利用CCK-8法检测Pseurata H对细胞增殖活性的影响；Griess法和ELISA试剂盒分别检测细胞上清液中NO、IL-6和TNF-α的含量；Western Blot检测COX-2、iNOS蛋白、p-NF-κB和p-ERK蛋白的表达变化；以RT-qPCR法检测药物干预前后细胞中iNOS、TNF-α、IL-6和IL-1β的mRNA基因表达变化。结果表明：浓度为10、20和40 μmol/L的Pseurata H对细胞没有毒性，不同浓度的Pseurata H对LPS诱导RAW264.7细胞炎症模型中NO、IL-6和TNF-α分泌具有很好的下调作用，对iNOS、COX-2、NF-κB、p-NF-κB、ERK和p-ERK蛋白的表达具有良好的抑制作用，且具有浓度效应。Pseurata H具有明显的抗炎作用，其作用机制可能是通过抑制 NF-κB/MAPK 信号通路而发挥抗炎作用。

关键词: 川藏香茶菜庚素, RAW264.7细胞, 脂多糖, 炎症, NF-κB/MAPK通路

Abstract: To study the anti-inflammatory effect and mechanism of Pseurata H on peritoneal macrophages (RAW264.7 cells) inflammation models, RAW264.7 cells were stimulated with LPS to establish an inflammation model in vitro, and Pseurata H with the concentrations of 10, 20, and 40 μmol/L was used to interfere with this cell model. The cytotoxicity of Pseurata H in RAW264.7 cells was determined by CCK-8 colorimetric assay. Griess assay was employed to detect the content of NO in the supernatant. The release of IL-6 and TNF-α in the supernatant was assayed using ELISA. The protein expressions of COX-2, iNOS, p-NF-κB and the p-ERK were measured by Western Blot. The mRNA expressions of iNOS, TNF-α, IL-6 and IL-1β were analyzed by RT-qPCR. The results showed that Pseurata H (10, 20, 40 μmol/L) displayed no apparent cytotoxicity in RAW264.7 cells. The release levels of NO, iNOS, TNF-α, and IL-6 were decreased by Pseurata H (10, 20, 40 μmol/L). In addition, Pseurata H also inhibited the expressions of iNOS, COX-2, NF-κB, p-NF-κB, ERK, and p-ERK proteins with concentration effect. Pseurata H has a significant anti-inflammatory effect, and its mechanism of action may be through the inhibition of NF-κB/MAPK signaling pathway to exert anti-inflammatory effects.

Key words: Pseurata H, RAW264.7 cells, lipopolysaccharide, inflammation, NF-κB/MAPK pathway

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