TY - JOUR
T1 - Magnoflorine Enhances LPS-Activated Pro-Inflammatory Responses via MyD88-Dependent Pathways in U937 Macrophages
AU - Haque, Md Areeful
AU - Jantan, Ibrahim
AU - Harikrishnan, Hemavathy
AU - Abdul Wahab, Siti Mariam
N1 - Funding Information:
The authors thank the Ministry of Agriculture and Agro-based Industries, Malaysia, for the financial support under the NKEA Research Grant Scheme (NRGS) (Grant no. NH1015D075).
Publisher Copyright:
© 2018 Georg Thieme. All rights reserved.
PY - 2018/6/15
Y1 - 2018/6/15
N2 - 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.
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.
KW - Akt
KW - immunomodulation
KW - magnoflorine
KW - MAPKs
KW - Menispermaceae
KW - MyD88
KW - NF- κ B
KW - Tinospora crispa
UR - http://www.scopus.com/inward/record.url?scp=85048854163&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85048854163&partnerID=8YFLogxK
U2 - 10.1055/a-0637-9936
DO - 10.1055/a-0637-9936
M3 - Article
C2 - 29906814
AN - SCOPUS:85048854163
SN - 0032-0943
VL - 84
SP - 1255
EP - 1264
JO - Planta Medica
JF - Planta Medica
IS - 17
ER -