Nutr Res Pract.  2011 Apr;5(2):101-106.

Anti-inflammatory effect of the water fraction from hawthorn fruit on LPS-stimulated RAW 264.7 cells

• Li C
• Wang MH

Abstract

The hawthorn fruit (Crataegus pinnatifida Bunge var. typica Schneider) is used as a traditional medicine in Korea. The objective of this study was to understand the mechanisms of the anti-inflammatory effects of the water fractionated portion of hawthorn fruit on a lipopolysaccharide (LPS)-stimulated RAW 264.7 cellular model. The level of nitric oxide (NO) production in the water fraction and LPS-treated RAW 264.7 cells were determined with an ELISA. The cytotoxicity of the water fraction and LPS was measured with an MTT assay. Expression of nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-alpha, interleukin 6 (IL-6), and interleukin 1beta (IL-1beta) mRNA were analyzed with a reverse transcription polymerase chain reaction (RT-PCR). The water fraction of hawthorn fruit was determined to be safe and significantly inhibited NO production in LPS-stimulated RAW 264.7 cells and suppressed COX-2, TNF-alpha, IL-1beta, and IL-6 expression. The observed anti-inflammatory effects of the water fraction of hawthorn fruit might be attributed to the down-regulation of COX-2, TNF-alpha, IL-1beta, and IL-6 expression in LPS-stimulated RAW 264.7 cells.

Keyword

Anti-inflammatory; hawthorn fruit; LPS; RAW 264.7 cell

MeSH Terms

Crataegus
Cyclooxygenase 2
Down-Regulation
Enzyme-Linked Immunosorbent Assay
Fruit
Interleukin-1beta
Interleukin-6
Korea
Medicine, Traditional
Nitric Oxide
Nitric Oxide Synthase
Polymerase Chain Reaction
Reverse Transcription
RNA, Messenger
Tumor Necrosis Factor-alpha
Water
Cyclooxygenase 2
Interleukin-1beta
Interleukin-6
Nitric Oxide
Nitric Oxide Synthase
RNA, Messenger
Tumor Necrosis Factor-alpha
Water

Figure

• 

  Fig. 1 Cell viability of murine macrophage RAW 264.7 cells. (A) Cell viability of RAW 264.7 cells after incubation in the presence of various concentrations of hawthorn fruit water fraction for 24 h. (B) Cell viability of RAW 264.7 cells co-treated with hawthorn fruit water fraction and 2 µg/ml of lipopolysaccharide (LPS) for 24 h. Cell viability was measured using an MTT assay. Data represent the mean ± SD of three independent experiments.

• 

  Fig. 2 Inhibitory effect of hawthorn fruit water fraction on nitric oxide (NO) production in a culture medium of LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 2 µg/ml of lipopolysaccharide (LPS) and hawthorn fruit water fraction for 24 h. The absorbance was determined at 550 nm with an ELISA. Data represent the mean ± SD of three independent experiments.

• 

  Fig. 3 Effects of hawthorn fruit water fraction on nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumour necrosis factor (TNF)-α, interleukin 6 (IL-6), and interleukin 1β (IL-1β) mRNA production in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 2 µg/ml of lipopolysaccharide (LPS) and various concentrations of hawthorn fruit water fraction for 24 h. (A) Reverse transcription polymerase chain reaction (RT-PCR) analysis of the expression of iNOS, COX-2, TNF-α, IL-1β, and IL-6 mRNA. (B), (C), (D), (E), and (F) Quantification of the iNOS, COX-2, TNF-α, IL-1β, and IL-6 expression levels were achieved with densitometric measurement.

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