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KMID : 1129720120290010037
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Korean Journal of Acupuncture
2012 Volume.29 No. 1 p.37 ~ p.46
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Pinus Densiflora Gnarl Extract for Pharmacopuncture Inhibits Inflammatory Responses through Heme Oxygenase-1 Induction in Lipopolysaccharide-Stimulated RAW264.7 Macrophages
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Lee Kang-Pa
Moon Jin-Young
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Abstract
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Objectives: The gnarl of Pinus densiflora, called Songjeol in Korea, has been used as a medicinal herb for the treatment of inflammatory-related diseases such as arthralgia, myalgia and bruise. However, the molecular actions and mechanisms have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of Pinus densiflora gnarl pharmacopuncture (PDGP) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells.
Methods: Cytotoxicity was assessed by XTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expressions of interleukin-1β (IL-1β), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) were analyzed by RT-PCR. Reactive oxidative species (ROS) generation was measured using the fluorescence microscopy. In addition, inducible nitric oxide synthase (iNOS) and redox factor-1 (Ref-1) protein expressions were detected by Western blotting.
Results: PDGP inhibited NO production and ROS generation in LPS-stimulated RAW264.7 cells. At the mRNA level, PDGP suppressed IL-1β, IL-6 and COX-2 expression. On the other hand, PDGP induced HO-1 mRNA expression. Furthermore, PDGP suppressed iNOS and Ref-1 protein expression.
Conclusions: This result suggests that PDGP can act as a suppressor agent on NO and iNOS through induction of HO-1, and play an useful role in blocking inflammatory responses.
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KEYWORD
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Pinus densiflora gnarl, pharmacopuncture, nitric oxide, heme oxygenase-1
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