Fighting Inflammation, The Body’s Silent Killer

What’s scarier than the boogeyman you know? The boogeyman you don’t
know. Well, chronic inflammation is indeed the boogeyman you don’t know because it isn’t felt until the pain or dysfunction from years of its silent reign inspire physician visits.

Cancer. Alzheimer’s. Autoimmune diseases. Depression. What do all these
conditions have in common? They have been linked to chronic inflammation. It seems impossible that one thing can be responsible for so many problems, but a growing body of medical research is revealing that long-term, systemic inflammation that occurs when one’s immune system goes into overdrive—sending out cells that attack healthy tissues, along with the unhealthy—may beat the root of many prevalent diseases. The medical and nutraceutical research communities are highly aware how chronic inflammation damages the human organism in many ways. In vivo research has outlined and clarified the cascade of reactions causing unhealthy inflammation.

Inflammation, however, is not an unjustified biological response. It is critical for our survival and is the body’s natural mechanism to defend against a diverse variety of pathogens including bacteria, viruses, fungi, tumors and other harmful agents (e.g., chemicals, radiation, burns and wounds). Inflammation is a complex reaction of the body in response to cellular injury (such as a bruise). It is marked by tissue swelling, capillary dilation, antihistamine activity, redness, heat and pain. It serves as a mechanism initiating the elimination of noxious agents and of damaged tissue (see Figure 1).

NF-kB: The Master Inflammation Switch

Implicated in the onset of 95% of all cancers is a common but under-recognized molecule most people have never heard of: nuclear factor-kappaB (NFkB). Emerging research validates the danger it poses to maturing individuals.

NF-kB is a protein that acts as a switch to turn inflammation on and off in the body. Scientists describe NF-kB as a “smoke sensor” that detects dangerous threats like free radicals and infectious agents. In response to these threats, NF-kB “turns on” genes that produce inflammation. As we age, NF-kB expression in the body increases, provoking widespread chronic inflammation and setting the stage for diseases ranging from atherosclerosis and diabetes to Alzheimer’s. Evidence from recent research suggests that by inhibiting NF-kB, a wide range of diseases and conditions in which inflammation plays a critical role can be treated (see Figure 2).

ParActin®: The Natural NF-kB Inhibitor

ParActin® helps balance and calm over-protective immune cells. Andrographolides in ParActin® can exert their benefi cial effects through PPAR gamma activation (i.e., PPAR gamma agonist), which effectively turns off the “master power switch” NF-kB, responsible for exerting the over-reactive inflammatory response. By deactivating NF-kB, the hyperexpression of cytokines, proinflammatory proteins and enzymes such as COX-2, PGE2, interleukin-2 and interferon gamma is reduced (see Figure 3). ParActin® is a patented extract from the plant called Andrographis paniculata, standardized to andrographolide, 14 deoxyandrographolide and neoandrographolide.

These naturally occurring phytochemicals in the Andrographis paniculata plant have been shown by researchers to have beneficial actions in supporting healthy joints, bones and muscles; this is so important in today’s active lifestyle. ParActin® was awarded U.S. Patent #8,084,495 B2 (Dec. 27, 2011), titled, “Composition of Labdane Diterpenes Extracted from Andrographis paniculata, Useful for the Treatment of Autoimmune Diseases, and Alzheimer’s Disease by Activation for PPAR-Gamma Receptors.”

Controlled Study 1

Andrographolide Inhibits IFN-g & IL-2 Cytokine Production and Protects Against Cell Apoptosis

Rafael A. Burgos, Karina Seguel, Mirna Perez, Ada Meneses, Marcela Ortega, Maria I. Guarda, Anitsi Loaiza, Juan L. Hancke

Controlled Study 2

Andrographolide interferes with binding of nuclear factor-kB to DNA in HL-60-derived neutrophilic cells,  Marı´a A. Hidalgo, Alex Romero, Jaime Figueroa, Patricia Corte´ s, Ilona I. Concha, Juan L. Hancke & , Rafael A. Burgos

Andrographolide is a labdane diterpene isolated from the
leaves of Andrographis paniculata, a Chinese herbal medicine
used for the treatment of viral infections and inflammatory
diseases (Coon & Ernst, 2004). Andrographolide is known to
exert several anti-inflammatory actions, including inhibition of
intercellular adhesion molecule-1 expression in monocytes
activated by tumor necrosis factor-a (Habtemariam, 1998),
suppression of inducible nitric oxide synthetase (iNOS)
expression in RAW264.7 cells stimulated by lipopolysaccharide
(LPS) and interferon-g (Chiou et al., 1998; 2000) and
inhibition of microglial activation through inhibition of iNOS
and cyclooxygenase-2 (COX-2) expression (Wang et al., 2004).
In neutrophils, andrographolide reduces adhesion induced by
N-formyl-methionyl-leucyl-phenylalanine (fMLP) (Shen et al.,
2000), and reactive oxygen species production and Mac-1 expression induced by phorbol myristate acetate and fMLP
(Shen et al., 2002).
Nuclear factor kappaB (NF-kB) is a transcription factor
found in a great variety of immune cells participating in the
regulation of genes involved in cellular and physiological
process, such as growth and apoptosis, and has an important
role in the inflammatory and immune response by inducing the
transcription of proinflammatory genes (Baeuerle & Baltimore,
1996). For instance, the proinflammatory mediators
such as intercellular adhesion molecule-1, iNOS and COX-2
are proteins regulated by the transcription factor NF-kB
(Barnes & Karin, 1997; Roebuck & Finnegan, 1999). Activation
of NF-kB involves the phosphorylation of specific
inhibitory factors (IkBa, IkBb and IkBe) by I-kappa kinase
(IKK)1, IKK2 and NF-kB essential modulator (Karin &
Delhase, 2000). Phosphorylated IkB is rapidly degraded in
response to stimuli (Simeonidis et al., 1999). This degradation
allows the resultant free NF-kB dimer to translocate into the
nucleus and induces gene transcription (Baldwin, 1996; Ghosh et al., 1998). Activation of NF-kB has an important
role in neutrophils, regulating apoptosis (Ward et al., 1999),
production of interleukin-1, -6, -8 and adhesion molecules
(Baeuerle & Henkel, 1994) and expression of COX-2 (Kim
et al., 2001).
In neutrophils, NF-kB is activated by proinflammatory
stimulus such as, fMLP and platelet-activating factor (PAF)
(McDonald et al., 1997). fMLP and PAF are factors that
mediate diverse neutrophil functions, such as chemotaxis,
superoxide production and COX-2 expression. These factors
bind to G-protein-coupled receptors, which induce mitogenactivated
protein kinase (MAPK) activation (Chao & Olson,
1993; Ishii & Shimizu, 2000). PAF activates p38 and
extracellular signal-regulated protein kinase (ERK)1/2 MAPK
in neutrophils (Nick et al., 1997; Hidalgo et al., 2004; Khreiss
et al., 2004). On the other hand, it has been demonstrated that
fMLP activates mainly ERK1/2 protein in human neutrophils
(Nick et al., 1997).
Several natural compounds have been reported to reduce
NF-kB activation (Bremner & Heinrich, 2002) and in the
present study, using neutrophil-like dimethylsulfoxide
(DMSO) differentiated HL-60 cells, as a cellular model of
neutrophils (Santos-beneit & Mollinedo, 2000), we have shown
that NF-kB activation by PAF and fMLP is strongly inhibited
by andrographolide, an effect that is mediated by andrographolide
blocking the binding of NF-kB to DNA.

Controlled Study 3

Andrographolide reduces IL-2 production in T-cells by interfering with NFAT and MAPK activation. María D. Carretta a, Pablo Alarcón a, Evelyn Jara a, Loreto Solis a, Juan L. Hancke a, Ilona I. Concha b, María A. Hidalgo a, Rafael A. Burgos a,⁎

The nuclear factor of activated Tcells (NFAT) is a transcription factor essential for cytokine production during T-cell
activation and is the target of several immunosuppressive drugs. Andrographolide is a diterpenic labdane that
possesses anti-inflammatory and immunomodulatory effects. Several studies propose that andrographolide can
reduce the immune response through inhibition of the nuclear factor kappa B (NF-κB) and mitogen-activated
protein kinases (MAPK) such as extracellular signal regulated kinase 1/2 (ERK1/2) pathways. Moreover,
andrographolide reduces IFN-γ and IL-2 production induced by concanavalin A inmurine T-cell.Nevertheless, the
mechanisms involved in the decrease of cytokine production are unknown. In the present study, we determined
that andrographolide reduced IL-2 production in Jurkat cells stimulated with phorbol myristate acetate and
ionomycin (PMA/Ionomycin). We then showed that andrographolide reduced NFAT luciferase activity and
interfered with its nuclear distribution, with these effects being linked to an increase in c-jun-N-terminal kinase
(JNK) phosphorylation. Additionally, reduction of NF-κB activity in Jurkat cells treatedwith andrographolidewas
observed. Using Western blotting, we demonstrated that andrographolide decreased ERK1 and ERK5
phosphorylation induced by anti-CD3 or PMA/Ionomycin. Andrographolide did not affect cell viability at
concentration of 10 and 50 μM; however, our results suggest that andrographolide increase early apoptosis at
100 μM. We concluded that andrographolide can exert immunomodulatory effects by interfering with NFAT
activation and ERK1 and ERK5 phosphorylation in T-cells.