Activation of murine macrophages by G1-4A, a polysaccharide from Tinospora cordifolia, in TLR4/MyD88 dependent manner

• Published in: International immunopharmacology 2017-09, Vol.50, p.168-177
• Main Authors: Gupta, Pramod Kumar, Rajan, M.G.R., Kulkarni, Savita
• Format: Article
• Language: English
• Subjects: Adjuvants, Immunologic; Animals; Antibodies; Antibodies, Blocking - metabolism; Blocking antibodies; Cell activation; Classical pathway; Cytokines; Cytokines - metabolism; Female; Herbal medicine; Immune response; Immune system; Immunity, Innate; Immunomodulator; Inflammation Mediators - metabolism; Innate immunity; Interferon; Interleukin 10; Interleukin 12; Interleukin 6; JNK protein; Macrophage; Macrophage Activation; Macrophages; Macrophages - immunology; Major histocompatibility complex; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; MyD88 protein; Myeloid Differentiation Factor 88 - genetics; Myeloid Differentiation Factor 88 - metabolism; Nitric oxide; Nitric Oxide Synthase Type II - genetics; Nitric Oxide Synthase Type II - metabolism; Peritoneum; Polysaccharides; Polysaccharides - immunology; RAW 264.7 Cells; RNA, Small Interfering - genetics; Saccharides; Signal Transduction; siRNA; Tinospora - immunology; Tinospora cordifolia; TLR4 protein; Toll-Like Receptor 4 - genetics; Toll-Like Receptor 4 - metabolism; Toll-like receptors; Tumor necrosis factor-α; Tumorigenesis; γ-Interferon
• ISSN: 1567-5769; 1878-1705
• DOI: 10.1016/j.intimp.2017.06.025

Macrophages are centrally placed in the innate immune system and their activation is crucial to the generation of appropriate immune response in the event of any pathogenic invasion, tumorigenesis or other human diseases. Many plant derived polysaccharides are known to activate macrophages. In the present study, effects of G1-4A, a polysaccharide derived from Tinospora cordifolia, on the activation of macrophages were investigated. Our data demonstrated the up regulation of expression of TNF-α, IL-β, IL-6, IL-12, IL-10 and IFN-γ in RAW 264.7 cell line and peritoneal macrophages after G-14A treatment. Nitric oxide levels were also enhanced along with up-regulation of NOS2 expression in murine macrophages post G1-4A treatment. Further, G1-4A treatment up-regulated the surface expression of MHC-II and CD-86 in macrophages. Using siRNA against TLR4, MyD88 and anti-TLR4 blocking antibodies, we established that G1-4A activated macrophages by classical pathway in TLR4-MyD88 dependent manner. Additionally, G1-4A treatment activated p38, ERK and JNK MAPKs in macrophages. Using pharmaceutical inhibitors of above MAPKs we concluded that G1-4A activates the macrophages by activation of p38, ERK and JNK MAPKs in RAW264.7 macrophages. Thus our data suggests the activation of macrophages by classical pathway after treatment of G1-4A. •G1-4A treatment activates murine macrophages through classical pathway.•G1-4A mediated macrophage activation is TLR4/MyD88 dependent.•ERK, JNK and p38 MAPK were involved in G1-4A mediated macrophage activation.