Immune Netw.  2010 Dec;10(6):212-218. 10.4110/in.2010.10.6.212.

The Stem Bark of Kalopanax pictus Exhibits Anti-inflammatory Effect through Heme Oxygenase-1 Induction and NF-kappaB Suppression

• Bang SY
• Park GY
• Park SY
• Kim JH
• Lee YK
• Lee SJ
• Kim Y

• KMID: 2150677
• DOI: http://doi.org/10.4110/in.2010.10.6.212

Abstract

BACKGROUND
The stem bark of Kalopanax pictus (KP) has been used in traditional medicine to treat rheumatoidal arthritis, neurotic pain and diabetes mellitus in China and Korea. In this study, the mechanism responsible for anti-inflammatory effects of KP was investigated.
METHODS
We examined the effects of KP on NO production, nitric oxide synthase (iNOS) and HO-1 expression, NF-kappaB, Nrf2 and MAPK activation in mouse peritoneal macrophages.
RESULTS
The aqueous extract of KP inhibited LPS-induced NO secretion as well as inducible iNOS expression, without affecting cell viability. KP suppressed LPS-induced NF-kappaB activation, phosphorylation and degradation of IkappaB-alpha, phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). Furthermore, KP induced HO-1 expression and Nrf2 nuclear translocation.
CONCLUSION
These results suggest that KP has the inhibitory effects on LPS-induced NO production in macrophages through NF-kappaB suppression and HO-1 induction.

Keyword

Kalopanax pictus; NO; NF-kappaB; MAPK; HO-1; Nrf2

MeSH Terms

Animals
Arthritis
Cell Survival
China
Diabetes Mellitus
Heme
Heme Oxygenase-1
I-kappa B Proteins
JNK Mitogen-Activated Protein Kinases
Kalopanax
Korea
Macrophages
Medicine, Traditional
Mice
NF-kappa B
Nitric Oxide Synthase
Phosphorylation
Phosphotransferases
Heme
Heme Oxygenase-1
I-kappa B Proteins
JNK Mitogen-Activated Protein Kinases
NF-kappa B
Nitric Oxide Synthase
Phosphotransferases

Figure

• 

  Figure 1 Effect of KP on the NO secretion and iNOS expression in TG-elicited mouse peritoneal macrophages. (A) Cells were incubated with various concentrations of KP for 1 h and then stimulated with 0.1µg/ml LPS for 24 h at 37℃. The amount of nitrite released was measured by the method of Griess. Values are means±S.E. of three independent experiments. *p<0.05 and †p<0.01 vs LPS-treated group. (B) Cells were treated with KP and/or LPS as mentioned above and equal cytosolic extracts were analyzed by Western blotting with anti-iNOS antibody. Western blot detection of β-tubulin was estimated protein-loading control for each lane. (C) Cells were incubated with various concentration of UR in presence of 0.1µg/ml of LPS for 24 h. Then cell viability was measured by MTT assay as described in Materials and Methods. Data represent the relative viability to control group and are expressed as the means±S.E. of three independent experiments.

• 

  Figure 2 Inhibitory effect of KP on NF-κB activity and phosphorylation/degradation of IκB-α. (A) Effect of KP on DNA binding activity of NF-κB. TG-elicited mouse peritoneal macrophages were incubated with various concentrations of KP for 1 h, and then stimulated with LPS (0.1µg/ml) for 30 min. Nuclear proteins were extracted and assayed for NF-κB DNA binding affinity by EMSA as described in Materials and methods. ns, non-specific band. (B) Effect of KP on LPS-stimulated phosphorylation and degradation of IκB-α. Cells were incubated with various concentrations of KP for 1 hr, and then stimulated with 0.1µg/ml LPS for 15 min. Cells were harvested and equal cytosolic extracts were analyzed by Western blotting with anti-IκB-α and -phospho-IκB-α. Western blot detection of β-tubulin was estimated protein-loading control for each lane.

• 

  Figure 3 Effect of KP on ERK1/2 and JNK activity in LPS-stimulated macrophages. TG-elicited mouse peritoneal macrophages were treated with indicated concentrations of KP for 1 h and stimulated with 0.1µ g/ml LPS for 15 min. Equal amount of cell extracts was analyzed by western blotting with anti-phospho-ERK1/2 or -phospho-JNK antibody. Western blot detection of non-phosphorylated kinases was estimated protein-loading control for each lane.

• 

  Figure 4 Effect of KP on HO-1 expression and nuclear translocation of in macrophages. (A) TG-elicited mouse peritoneal macrophages were incubated with indicated concentrations of KP for 24 h. HO-1 protein levels of cytosolic extracts were analyzed by Western blotting. (B) Cells were incubated with indicated concentrations of KP for 3 h and the levels of Nrf2 in nuclear extracts were analyzed by Western blotting. Western blot detection of β-tubulin was estimated protein-loading control for each lane.

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