Magnoflorine Enhances LPS-Activated Pro-Inflammatory Responses via MyD88-Dependent Pathways in U937 Macrophages

Md Areeful Haque, Ibrahim Jantan, Hemavathy Harikrishnan, Siti Mariam Abdul Wahab

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Magnoflorine, a major bioactive metabolite isolated from Tinospora crispa, has been reported for its diverse biochemical and pharmacological properties. However, there is little report on its underlying mechanisms of action on immune responses, particularly on macrophage activation. In this study, we aimed to investigate the effects of magnoflorine, isolated from T. crispa on the pro-inflammatory mediators generation induced by LPS and the concomitant NF- κ B, MAPKs, and PI3K-Akt signaling pathways in U937 macrophages. Differentiated U937 macrophages were treated with magnoflorine and the release of pro-inflammatory mediators was evaluated through ELISA, while the relative mRNA expression of the respective mediators was quantified through qRT-PCR. Correspondingly, western blotting was executed to observe the modulatory effects of magnoflorine on the expression of various markers related to NF- κ B, MAPK and PI3K-Akt signaling activation in LPS-primed U937 macrophages. Magnoflorine significantly enhanced the upregulation of TNF- α , IL-1 β , and PGE 2 production as well as COX-2 protein expression. Successively, magnoflorine prompted the mRNA transcription level of these pro-inflammatory mediators. Magnoflorine enhanced the NF- κ B activation by prompting p65, I κ B α , and IKK α / β phosphorylation as well as I κ B α degradation. Besides, magnoflorine treatments concentration-dependently augmented the phosphorylation of JNK, ERK, and p38 MAPKs as well as Akt. The immunoaugmenting effects were further confirmed by investigating the effects of magnoflorine on specific inhibitors, where the treatment with specific inhibitors of NF- κ B, MAPKs, and PI3K-Akt proficiently blocked the magnoflorine-triggered TNF- α release and COX-2 expression. Magnoflorine furthermore enhanced the MyD88 and TLR4 upregulation. The results suggest that magnoflorine has high potential on augmenting immune responses.

Original languageEnglish
JournalPlanta Medica
DOIs
Publication statusAccepted/In press - 15 Jun 2018

Fingerprint

Macrophages
Phosphatidylinositol 3-Kinases
Phosphorylation
Chemical activation
magnoflorine
Up-Regulation
Tinospora
Messenger RNA
Macrophage Activation
p38 Mitogen-Activated Protein Kinases
Transcription
Metabolites
Prostaglandins E
Interleukin-1
Western Blotting
Enzyme-Linked Immunosorbent Assay
Pharmacology

Keywords

  • Akt
  • immunomodulation
  • magnoflorine
  • MAPKs
  • Menispermaceae
  • MyD88
  • NF- κ B
  • Tinospora crispa

ASJC Scopus subject areas

  • Analytical Chemistry
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Drug Discovery
  • Complementary and alternative medicine
  • Organic Chemistry

Cite this

Magnoflorine Enhances LPS-Activated Pro-Inflammatory Responses via MyD88-Dependent Pathways in U937 Macrophages. / Haque, Md Areeful; Jantan, Ibrahim; Harikrishnan, Hemavathy; Abdul Wahab, Siti Mariam.

In: Planta Medica, 15.06.2018.

Research output: Contribution to journalArticle

Haque, Md Areeful ; Jantan, Ibrahim ; Harikrishnan, Hemavathy ; Abdul Wahab, Siti Mariam. / Magnoflorine Enhances LPS-Activated Pro-Inflammatory Responses via MyD88-Dependent Pathways in U937 Macrophages. In: Planta Medica. 2018.
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AU - Jantan, Ibrahim

AU - Harikrishnan, Hemavathy

AU - Abdul Wahab, Siti Mariam

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AB - Magnoflorine, a major bioactive metabolite isolated from Tinospora crispa, has been reported for its diverse biochemical and pharmacological properties. However, there is little report on its underlying mechanisms of action on immune responses, particularly on macrophage activation. In this study, we aimed to investigate the effects of magnoflorine, isolated from T. crispa on the pro-inflammatory mediators generation induced by LPS and the concomitant NF- κ B, MAPKs, and PI3K-Akt signaling pathways in U937 macrophages. Differentiated U937 macrophages were treated with magnoflorine and the release of pro-inflammatory mediators was evaluated through ELISA, while the relative mRNA expression of the respective mediators was quantified through qRT-PCR. Correspondingly, western blotting was executed to observe the modulatory effects of magnoflorine on the expression of various markers related to NF- κ B, MAPK and PI3K-Akt signaling activation in LPS-primed U937 macrophages. Magnoflorine significantly enhanced the upregulation of TNF- α , IL-1 β , and PGE 2 production as well as COX-2 protein expression. Successively, magnoflorine prompted the mRNA transcription level of these pro-inflammatory mediators. Magnoflorine enhanced the NF- κ B activation by prompting p65, I κ B α , and IKK α / β phosphorylation as well as I κ B α degradation. Besides, magnoflorine treatments concentration-dependently augmented the phosphorylation of JNK, ERK, and p38 MAPKs as well as Akt. The immunoaugmenting effects were further confirmed by investigating the effects of magnoflorine on specific inhibitors, where the treatment with specific inhibitors of NF- κ B, MAPKs, and PI3K-Akt proficiently blocked the magnoflorine-triggered TNF- α release and COX-2 expression. Magnoflorine furthermore enhanced the MyD88 and TLR4 upregulation. The results suggest that magnoflorine has high potential on augmenting immune responses.

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