Gypenoside XLIX isolated from Gynostemma pentaphyllum inhibits nuclear factor-kappaB activation via a PPAR-alpha-dependent pathway

Summary

Nuclear factor (NF)-κB is important in the generation of inflammation. Besides regulating lipid metabolism, peroxisome proliferator-activated receptor (PPAR)-α activators also reduce NF-κB activation to terminate activation of inflammatory pathways. Gynostemma pentaphyllum (GP) has been used to treat various inflammatory diseases and hyperlipidemia. Here, we demonstrate that GP extract and one of its main components, Gypenoside XLIX (Gyp-XLIX) inhibited LPS-induced NF-κB activation in murine macrophages. Furthermore, Gyp-XLIX restored the LPS- and TNF-α-induced decrease in cytosolic I-κBα protein expression and inhibited the translocation of NF-κB(p65) to the nucleus in THP-1 monocyte and HUVEC cells. The inhibition of LPS- and TNF-α-induced NF-κB luciferase activity in macrophages was abolished by MK-886, a selective PPAR-α antagonist. GP extract and Gyp-XLIX (EC50: 10.1 μM) enhanced PPAR-α luciferase activity in HEK293 cells transfected with the tK-PPREx3-Luc reporter plasmid and expression vectors for PPAR-α. Additionally, Gyp-XLIX specifically enhanced PPAR-α mRNA and protein expression in THP-1-derived macrophage cells. The selectivity of Gyp-XLIX for PPAR-α was demonstrated by the activation of only PPAR-α in HEK293 cells transfected with expression vectors for PPAR-α, PPAR-β/δ or PPAR-γ1 plasmids and in THP-1-derived macrophage naturally expressing all three PPAR isoforms. The present study demonstrates that Gyp-XLIX, a naturally occurring gynosaponin, inhibits NF-κB activation via a PPAR-α-dependent pathway.

Abbreviations

BSA:

bovine serum albumin

EDTA:

ethylenediaminetetraacetic acid

GP:

Gynostemma pentaphyllum

Gyp-XLIX:

gypenoside XLIX

HEK293:

human embryonic kidney 293

HUVEC:

human umbilical vein endothelial cells

iNOS:

inducible nitric oxide synthase

I-κB:

inhibitor-κB

LPS:

lipopolysaccharide

NF-κB:

nuclear factor-kappaB

NO:

nitric oxide

PBS:

phosphate-buffered saline

PDTC:

pyrrolidine dithiocarbamate

PPAR:

peroxisome proliferator-activated receptor

RT-PCR:

reverse transcription polymerase chain reaction

SDS:

sodium dodecyl sulfate

TNF-α:

tumor necrosis factor-α

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Acknowledgements

The authors wish to thank Ankang Pharmaceutical Company Beijing, China, for its generous gift of gypenosides extract, and Dr Rujee K. Duke and Mr Bruce N. Tattam for their assistance in the project.

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Correspondence to Basil D. Roufogalis.

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Tom Hsun-Wei Huang and Yuhao Li contributed equally.

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Huang, T.H., Li, Y., Razmovski-Naumovski, V. et al. Gypenoside XLIX isolated from Gynostemma pentaphyllum inhibits nuclear factor-kappaB activation via a PPAR-alpha-dependent pathway. J Biomed Sci 13, 535–548 (2006). https://doi.org/10.1007/s11373-006-9076-8

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Key words

  • Gynostemma pentaphyllum
  • gypenoside
  • nuclear factor-kappaB
  • peroxisome proliferator-activated receptor-alpha