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Pinus Densiflora Gnarl Extract for Pharmacopuncture Inhibits Inflammatory Responses through Heme Oxygenase-1 Induction in Lipopolysaccharide-Stimulated RAW264.7 Macrophages  

Lee, Kang-Pa (Division of Bio Science, College of Science and Technology, Dongguk University)
Moon, Jin-Young (Dept. of Meridian & Acupoint, College of Oriental Medicine, Dongguk University)
Publication Information
Korean Journal of Acupuncture / v.29, no.1, 2012 , pp. 37-46 More about this Journal
Abstract
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-$1{\beta}$ (IL-$1{\beta}$), 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-$1{\beta}$, 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.
Keywords
Pinus densiflora gnarl; pharmacopuncture; nitric oxide; heme oxygenase-1;
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