[논문]Anti-Inflammatory Activity of the Solvent-Partitioned Fractions from Spergularia marina in LPS-Stimulated RAW 264.7 Cells

Kong, Chang-Suk

doi:10.3746/pnf.2014.19.4.261

Anti-Inflammatory Activity of the Solvent-Partitioned Fractions from Spergularia marina in LPS-Stimulated RAW 264.7 Cells 원문보기

Preventive nutrition and food science, v.19 no.4, 2014년, pp.261 - 267

Kong, Chang-Suk (Department of Food and Nutrition, College of Medical and Life Science, Silla University)

Abstract ▼ AI-Helper

As a part of ongoing research to elucidate and characterize antioxidant and anti-inflammatory nutraceuticals, solvent-partitioned fractions from Spergularia marina were tested for their ability to scavenge radicals and suppress inflammation. The results of the 2',7'-dichlorofluorescein diacetate assay indicate that solvent-partitioned fractions from S. marina scavenged intracellular radicals in $H_2O_2$-stimulated mouse macrophages. The tested fractions decreased the generation of nitric oxide (NO) and the expression of inflammation mediators, namely, inducible nitric oxide synthase (iNOS) and interleukin (IL)-6, by lipopolysaccharide (LPS)-induced mouse macrophages, indicating that S. marina decreases inflammation. Among all tested fractions [i.e., $H_2O$, n-buthanol (n-BuOH), 85% aqueous methanol (aq. MeOH), and n-hexane], the 85% aq. MeOH fraction showed the strongest antioxidant and anti-inflammatory response. The 85% aq. MeOH fraction scavenged 80% of the free radicals produced by $H_2O_2$-induced control cells. In addition, NO production was 98% lower in 85% aq. MeOH fraction-treated cells compared to LPS-induced control cells. The mRNA expression of iNOS and IL-6 was also suppressed in 85% aq. MeOH fraction-treated cells. The results of the current study suggest that the phenolic compound components of S. marina are responsible for its antioxidant and anti-inflammatory effects.

주제어

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제안 방법

After confirming the anti-oxidative effect of the S. marina solvent fractions, the anti-inflammatory potential of these fractions was evaluated by assessing NO production by LPS-stimulated RAW 264.7 cells in the presence and absence of the solvent-partitioned S. marina fractions. The presence of LPS activated the RAW 264.
The cells were pretreated with LPS (1 μg/mL), followed by treatment with the solvent fractions for 24 h (A) and 48 h (B). The nitrite content of the culture media was analyzed. Means with different letters (a-e) are significantly different (P<0.
Strong antioxidant molecules are considered to show anti-inflammatory effect due to close relation between inflammatory and oxidative stress pathways (8). Therefore, the anti-inflammatory activity of solvent-partitioned fractions from S. marina on LPS-stimulated RAW 264.7 mouse macrophages was evaluated in this study. When activated with pathogenic substances (e.

대상 데이터

The target cDNA was amplified using the following sense and antisense primers: forward 5'-TTCCAG-AAT-CCC-TGG-ACA-AG-3' and reverse 5'-TGGTCA-AAC-TCT-TGG-GGT-TC-3' for inducible nitric oxide synthase (iNOS); forward 5'-AGT-TGC-CTT-CTT-GGGACT-GA-3' and reverse 5'-CAG-AAT-TGC-CAT-TGCACA-AC-3' for interleukin (IL)-6; and forward 5'-CCACAG-CTG-AGA-GGG-AAA-TC-3' and reverse 5'-AAGGAA-GGC-TGG-AAA-AGA-GC-3' for β-actin.

데이터처리

Differences between the means of the individual groups were analyzed with the analysis of variance (ANOVA) procedure of Statistical Analysis System SAS v9.1 (SAS Institute, Cary, NC, USA) with Duncan’s multiple range tests.
After 15 min of incubation at room temperature, the absorbance was measured at 550 nm with a GENios^Ⓡ microplate reader (Tecan Austria GmbH). Nitrite concentrations were calculated by regression analysis. Serial dilutions of sodium nitrite were used as a standard.

이론/모형

Relative quantification was calculated using the 2−ΔΔCT method.
The total polyphenol content of S. marina fractions was determined by the Folin-Ciocalteu method. In brief, 20 μL of each extract was mixed with 100 μL of 1:10 FolinCiocalteu reagent in a microplate.

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