Activation of RAW264.7 macrophages by an acidic polysaccharide derived from Citrus grandis 'Tomentosa'

Int J Biol Macromol. 2020 Aug 1:156:1323-1329. doi: 10.1016/j.ijbiomac.2019.11.172. Epub 2019 Nov 21.

Abstract

Citrus grandis 'Tomentosa' which is a special Citrus cultivar, has been employed as cough suppressant and expectorant in traditional Chinese medicine for thousands of years. The aim of this study is to investigate the immunomodulatory role of an acidic polysaccharide (designated as CGTP-AP) purified from C. grandis 'Tomentosa'. CGTP-AP showed effective immune activation in RAW264.7 macrophages at the concentration of 1-100 μg/mL. CGTP-AP could promote the release of NO in dose- and time-dependent manners. Enzyme-Linked Immunosorbent Assay (ELISA) and RT-PCR analysis demonstrated that CGTP-AP could stimulate the secretion of TNF-α and IL-6 in a dosage-dependent way. Western blot analysis and RT-PCR analysis indicated that CGTP-AP treatment could induce the iNOS and COX-2 expression in RAW264.7 macrophages. By conducting the inhibitors experiments, the activation of NF-κB and MAPK signaling pathways by CGTP-AP treatment was confirmed. Therefore, the present results declared that CGTP-AP could be a promising candidate as a potent immunomodulator for the application in future pharmaceutical development.

Keywords: Citrus grandis ‘Tomentosa’; Cytokines; Immune activation; Macrophages; Polysaccharides; Western blot.

MeSH terms

  • Animals
  • Citrus / chemistry*
  • Cyclooxygenase 2 / metabolism
  • Dose-Response Relationship, Drug
  • Gene Expression Regulation, Enzymologic / drug effects
  • Interleukin-6 / metabolism
  • MAP Kinase Signaling System / drug effects
  • Macrophage Activation / drug effects*
  • Macrophages / drug effects*
  • Macrophages / immunology*
  • Macrophages / metabolism
  • Mice
  • NF-kappa B / metabolism
  • Nitric Oxide Synthase Type II / metabolism
  • Polysaccharides / chemistry
  • Polysaccharides / pharmacology*
  • RAW 264.7 Cells
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Interleukin-6
  • NF-kappa B
  • Polysaccharides
  • Tumor Necrosis Factor-alpha
  • Nitric Oxide Synthase Type II
  • Cyclooxygenase 2