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A novel role of andrographolide, an NF-kappa B inhibitor, on inhibition of platelet activation: the pivotal mechanisms of endothelial nitric oxide synthase/cyclic GMP

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Abstract

Andrographolide is a novel NF-κB inhibitor from the leaves of Andrographis paniculata. Platelet activation is relevant to a variety of thrombotic diseases. However, no data are available concerning the effects of andrographolide in platelet activation. The aim of this study was to examine the mechanisms of andrographolide in preventing platelet activation. Andrographolide (25–75 μΜ) exhibited a more potent activity of inhibiting platelet aggregation stimulated by collagen. Andrographolide inhibited collagen-stimulated platelet activation accompanied by relative Ca2+ mobilization; thromboxane A2 formation; and phospholipase C (PLC)γ2, protein kinase C (PKC), mitogen-activated protein kinase (MAPK), and Akt phosphorylation. Andrographolide markedly increased cyclic GMP, but not cyclic AMP levels. Andrographolide also stimulated endothelial nitric oxide synthase (eNOS) expression, NO release, and vasodilator-stimulated phosphoprotein (VASP) phosphorylation. ODQ, an inhibitor of guanylate cyclase, markedly reversed the andrographolide-mediated inhibitory effects on platelet aggregation, p38 MAPK and Akt phosphorylation, and the andrographolide-mediated stimulatory effect on VASP phosphorylation. Furthermore, a PI3 kinase inhibitor (LY294002) but not a PKC inhibitor (Ro318220) significantly diminished p38 MAPK phosphorylation; nevertheless, a p38 MAPK inhibitor (SB203580) and LY294002 diminished PKC activity stimulated by collagen. Andrographolide also reduced collagen-triggered hydroxyl radical (OH⋅) formation. In vivo studies revealed that andrographolide (22 and 55 μg/kg) is effective in reducing the mortality of ADP-induced acute pulmonary thromboembolism and significantly prolonged platelet plug formation in mice. This study demonstrates for the first time that andrographolide possesses a novel role of antiplatelet activity, which may involve the activation of the eNOS-NO/cyclic GMP pathway, resulting in the inhibition of the PI3 kinase/Akt-p38 MAPK and PLCγ2–PKC cascades, thereby leading to inhibition of platelet aggregation.

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Acknowledgments

This work was supported by grants from the National Science Council of Taiwan (NSC94-2321-B-038-001 and NSC97-2320-B-038-016-MY3) and the Committee on Chinese Medicine and Pharmacy (CCMP97-RD-008).

Disclosure of potential conflict of interests

The authors declare no conflict of interests related to this study.

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Authors and Affiliations

  1. Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan

    Wan-Jung Lu, Thanasekaran Jayakumar, Tsorng-Han Fong & Joen-Rong Sheu

  2. Department of Surgery, Mackay Memorial Hospital, Taipei, Taiwan

    Jie-Jen Lee

  3. Department of Pharmacology, School of Medicine, College of Medicine, Taipei Medical University, 250 Wuxing St., Taipei, 11031, Taiwan

    Duen-Suey Chou, George Hsiao & Joen-Rong Sheu

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  1. Wan-Jung Lu
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Correspondence to Joen-Rong Sheu.

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Lu, WJ., Lee, JJ., Chou, DS. et al. A novel role of andrographolide, an NF-kappa B inhibitor, on inhibition of platelet activation: the pivotal mechanisms of endothelial nitric oxide synthase/cyclic GMP. J Mol Med 89, 1261–1273 (2011). https://doi.org/10.1007/s00109-011-0800-0

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