天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (10): 1657-1665.doi: 10.16333/j.1001-6880.2022.10.003

• 研究论文 • 上一篇    下一篇

太子参参须皂苷对RAW 264.7细胞体外免疫调节作用研究

曾  丽1,陈赛红1,甘思言2,杜蓥蓥1,张炎达3,黄一帆2*,马玉芳1*   

  1. 1中西兽医结合与动物保健福建省高等学校重点实验室;2福建省兽医中药与动物保健重点实验室,福州 350002;3 福建贝迪药业有限公司,宁德 355399
  • 出版日期:2022-10-28 发布日期:2022-10-31
  • 基金资助:
    福建省产学研重大专项(2014N5004)

Immuomodulatory effects of Radix Pseudostellariae fibrous root saponins on RAW 264.7 cells in vitro

ZENG Li1,CHEN Sai-hong1,GAN Si-yan2,DU Ying-ying1,ZHANG Yan-da3,HUANG Yi-fan2*,MA Yu-fang1*    

  1. 1University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province;2Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health,Fuzhou 350002,China;3 Fujian Bedi Pharmaceutical Co.,Ltd.,Ningde 355399,China
  • Online:2022-10-28 Published:2022-10-31

摘要:

本实验旨在研究太子参参须皂苷(Radix Pseudostellariae fibrous roots saponins,RPFRS)对小鼠单核巨噬细胞(RAW 264.7细胞)的免疫调节作用及可能的作用机制。采用脂多糖(LPS)刺激RAW 264.7细胞,检测RAW 264.7细胞的增殖、吞噬活性、NO产生量、相关细胞因子含量、相关免疫分子及免疫蛋白相对表达量。结果表明:与空白对照组相比,RPFRS组中的RAW 264.7细胞的吞噬活性OD值呈剂量依赖式上升(P<0.01),NO、IL-6的含量、IL-6和TNF-α的mRNA相对表达量呈剂量依赖式上升(P<0.05); RPFRS的浓度为12.5~100 μg/mL时, IL-10、IL-1β、TNF-α的含量及IL-10、IL-1β、iNOS、MyD88和NF-κB的mRNA表达量在呈剂量依赖式上升(P<0.05),但TLR4的mRNA相对表达量呈剂量依赖式降低(P<0.05);WB结果显示RPFRS能够上调TLR2、MyD88、TRIF与胞核NF-κB蛋白,下调TLR4、IκB-α和胞浆NF-κB蛋白的相对表达量,上述结果表明,RPFRS可以通过提高RAW 264.7细胞的增殖能力、吞噬活性、分泌NO和细胞因子能力从而增强RAW 264.7细胞的免疫功能,提示RPFRS可能通过TLR2/4-MyD88/TRIF-NF-κB信号通路活化RAW 264.7细胞。

关键词: 太子参参须皂苷, RAW 264.7细胞, 免疫调节

Abstract:

To study the immunomodulatory effects of Radix Pseudostellariae fibrous root saponins (RPFRS) and their mechanisms,we examined the effects of RPFRS in RAW 264.7 cells on the proliferation,phagocytic activity,production of NO and cytokines,the mRNA expression of immune molecules and relevant immune protein expression levels.RAW 264.7 cells were treated with different concentrations of RPFRS.A DMEM blank control group (control group) and LPS positive group (LPS group) were set at the same time.The results showed that compared with the control group,the OD values on phagocytosis of neutral red (P<0.01),the contents of NO,IL-6 and the mRNA expression levels of IL-6,TNF-α in the RPFRS groups were significantly increased in a dose-dependent manner.The contents of IL-10,IL-1β,TNF-α and the mRNA expression levels of IL-10,IL-1β,iNOS,MyD88 and NF-κB increased significantly in the RPFRS groups (P<0.05) when the RPFRS concentration was 12.5-100 μg/mL,but the mRNA expression of TLR4 decreased in a dose-dependent manner (P<0.05).At the same time,Western blotting results also showed that RPFRS can increase the expression of TLR2,MyD88,TRIF,and nuclear NF-κB protein,while the expression of TLR4,IκB-α and cytosolic NF-κB protein was decreased. Above results indicated that RPFRS could enhance the immunity of RAW 264.7 cells by increasing their proliferation capacity, phagocytic activity and the ability to secrete NO and cytokines. And the mechanism of immune recombination of RPFRS on RAW 264.7 cells might be through the TLR2/4-MyD88/TRIF-NF-κB pathway to activate RAW264.7 cells. 

Key words: Radix Pseudostellariae fibrous root saponins, RAW 264.7 cells, immunomodulation

中图分类号:  R285