[논문]Anti-inflammatory Effect of Dangyuja (Citrus grandis Osbeck) Leaves in LPS-stimulated RAW 264.7 Cells

Yang, Eun-Jin; Lee, Hye-Ja; Kang, Gyeoung-Jin; Park, Sun-Soon; Yoon, Weon-Jong; Kang, Hee-Kyoung; Cho, So-Mi Kim; Yoo, Eun-Sook

[국내논문] Anti-inflammatory Effect of Dangyuja (Citrus grandis Osbeck) Leaves in LPS-stimulated RAW 264.7 Cells 원문보기

Food science and biotechnology, v.18 no.5, 2009년, pp.1063 - 1070

Yang, Eun-Jin (Department of Pharmacology, College of Medicine, Jeju National University) , Lee, Hye-Ja (Department of Pharmacology, College of Medicine, Jeju National University) , Kang, Gyeoung-Jin (Department of Pharmacology, College of Medicine, Jeju National University) , Park, Sun-Soon (Department of Pharmacology, College of Medicine, Jeju National University) , Yoon, Weon-Jong (Department of Pharmacology, College of Medicine, Jeju National University) , Kang, Hee-Kyoung (Department of Pharmacology, College of Medicine, Jeju National University) , Cho, So-Mi Kim (Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University) , Yoo, Eun-Sook (Department of Pharmacology, College of Medicine, Jeju National University)

Abstract ▼ AI-Helper

Dangyuja (Citrus grandis Osbeck) is a native plant growing only on Jeju Island in Korea. In this study, antiinflammatory effect of dangyuja leaves on a murine macrophage cell line was investigated. RAW 264.7 murine macrophage cells were stimulated with lipopolysaccharide (LPS, $1{\mu}g/mL$) to induce expression of pro-inflammatory markers [interleukin (IL)-6 and inducible nitric oxide synthase (iNOS)]. The crude extract (80% MeOH Ex.) and solvent fractions (hexane, $CHCl_3$, EtOAc, BuOH, and $H_2O$ Ex.) were obtained from dangyuja leaves. The $CHCl_3$ fraction inhibited the nitric oxide (NO) and IL-6 production in a dose-dependent manner. Also, the $CHCl_3$ fraction inhibited mRNA expression and protein levels of iNOS in a dose-dependent manner. Furthermore, the $CHCl_3$ fraction inhibited LPS-induced nuclear factor (NF)-${\kappa}B$ activation and phosphorylation of mitogen-activated protein kinases (MAPKs: ERK, JNK, and p38). These results suggest that dangyuja leaves may inhibit LPS-induced production of inflammatory markers by blocking NF-${\kappa}B$ and MAPKs signaling in RAW 264.7 cells.

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가설 설정

To assess whether the tested fractions of dangyuja affected cell viability, RAW 264.7 cells were incubated with LPS (1 µg/mL) in the presence of fraction of dangyuja leaves (100 µg/mL).

제안 방법

In order to investigate the effect of dangyuja leaves on the NO production, we measured NO concentrations in the culture medium. We incubated RAW 264.
7 cells (32,33). Thus, we examined the effects of the CHCl₃ fraction on IL-6 production in LPSstimulated RAW 264.7 cells using the ELISA and RT-PCR. The CHCl₃ fraction inhibited IL-6 production and IL-6 mRNA expression in LPS stimulated RAW 264.

대상 데이터

Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Lipopolysaccharide (LPS, Escherichia coli 0111:B4) was purchased from SigmaAldrich (St. Louis, MO, USA). All other chemicals used were analytical grade.
Murine macrophage RAW 264.7 cells were purchased from the Korean Cell Line Bank (Seoul, Korea). They were cultured in DMEM containing 2 mM glutamine, 10 mM HEPES, penicillin (100 units/mL), streptomycin (100 µg/mL), and 10% FBS.

데이터처리

Student’s t-test was used to assess the statistical significance of differences.

이론/모형

Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (25,26). RAW 264.

성능/효과

In conclusion, we demonstrated that the CHCl3 fraction of dangyuja leaves markedly inhibits the expression of macrophage-mediated inflammation factors such as NO and IL-6 without cell cytotoxicity via a blockade of NF-κB activation and MAPKs (ERK1/2, JNK, and p38) phosphorylation in LPS-stimulated RAW 264.7 cells.

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