KMID : 1149220170250010001
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Korean Journal of Oriental Medical Prescription 2017 Volume.25 No. 1 p.1 ~ p.10
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Inhibition of NO Production by Ethanol Extract of Prunus mume Fruits in LPS-Stimulated RAW 264.7 Macrophages through Regulation of the Nrf2/HO-1 Signaling Pathway
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Kang Hye-Joo
Choi Eun-Ok Jeong Jin-Woo Park Shin-Hyung Park Cheol Hong Su-Hyun Shin Soon-Shik Cheong Jae-Hun Choi Yung-Hyun
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Abstract
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Objectives : The fruit of Prunus mume Siebold & Zucc. has been used as an alternative medicine and functional food in Korea and Japan for preventive and therapeutic purposes. However, its molecular actions and mechanism on anti-inflammatory activity have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of the ethanol extract of P. mume fruit (EEPM) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and sought to understand the associated molecular mechanisms.
Methods : Cytotoxicity was assessed by an MTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expression of inducible nitric oxide synthase (iNOS) was analyzed by RT-PCR. In addition, expression levels of iNOS, nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) protein were detected by Western blotting.
Results : Our data indicated that EEPM inhibited NO production in LPS-stimulated RAW264.7 cells in a concentration-dependent manner. At the mRNA and protein levels, EEPM suppressed LPS-induced iNOS expression. On the other hand, EEPM markedly enhanced HO-1 expression, which was associated with an induction and nuclear translocation of Nrf2. Moreover, the inhibitory effect of EEPM against LPS?induced NO production was significantly enhanced by hemin, a HO-1 inducer; however, EEPM's effect on the production of NO was abolished by zinc protoporphyrin IX, a HO-1 inhibitor.
Conclusion : The results suggest that EEPM can act as a suppressor agent on NO production through an activation of Nrf2/HO-1 signaling pathway, and may be a promising candidate for the treatment of inflammatory diseases.
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KEYWORD
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Prunus mume , NO , HO-1 , Nrf2 , anti-inflammatory activity
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