Polyphenol-rich pomegranate fruit extract (POMx) suppresses PMACI-induced expression of pro-inflammatory cytokines by inhibiting the activation of MAP Kinases and NF-kappaB in human KU812 cells.
BACKGROUND
Mast cells and basophils are multifunctional effector cells and contain plentiful secretary granules in their cytoplasm. These cell types are involved in several inflammatory and immune events and are known to produce an array of mediators including a broad spectrum of cytokines. Pomegranate fruit is rich in anthocyanins and hydrolysable tannins; a group of polyphenolic compounds shown to be potent antioxidant with anti-inflammatory activity. However, no studies have been undertaken to investigate whether a polyphenol-rich pomegranate fruit extract (POMx) inhibits the inflammatory activity of activated human mast cells and basophils. The aim of this study was to examine whether POMx modulates inflammatory reactions using human basophilic cell line KU812.


METHODS
KU812 cells were stimulated with phorbol-12-myristate 13-acetate plus calcium inophore A23187 (PMACI). The inhibitory effect of POMx on pro-inflammatory cytokine gene expression and production by stimulated KU812 cells was measured by quantitative RT-PCR, and cytokine-specific ELISA assays, respectively. Western blotting was used to analyze the effect of POMx on the activation of mitogen-activated protein kinases (MAPKs), and the nuclear factor (NF)-kappaB in PMACI stimulated KU812 cells. Effect on the activity of NF-kappaB was determined using Luciferase reporter assay. Significance of differences from control values were analyzed by means of standard statistical methods.


RESULTS
POMx significantly decreased PMACI stimulated inflammatory gene expression and production of interleukin (IL)-6 and IL-8 in KU812 cells. The inhibitory effect of POMx on the pro-inflammatory cytokines was MAPK subgroups c-jun N-terminal kinase (JNK)- and extracellular-regulated kinase (ERK) dependent. In addition, POMx suppressed the NF-kappaB activation induced by PMACI by inhibiting IkappaB-degradation in human basophil cells. POMx also suppressed the powerful induction of NF-kappaB promoter-mediated luciferase activity in transiently transfected KU812 cells.


CONCLUSIONS
These novel pharmacological actions of POMx provide new suggestion that POMx or POMx-derived compounds may be of therapeutic use for the treatment of inflammatory diseases by suppressing mast cells/basophils activation.
Rasheed
Zafar
Z
Department of Pathology, Microbiology, & Immunology, School of Medicine, University of South Carolina, 6439 Garners Ferry Road, Columbia, SC-29209, USA. Tariq.Haqqi@uscmed.sc.edu.
Akhtar
Nahid
N
Anbazhagan
Arivarasu N
AN
Ramamurthy
Sangeetha
S
Shukla
Meenakshi
M
Haqqi
Tariq M
TM