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Anti-inflammatory effect of the taffy mu yeot, made from the Korean radish Raphanus sativus L. in a lipopolysaccharide-induced murine model of pulmonary inflammation

Dan-Hee Kim
Dan-Hee Kim
 and 
Sang-Yun Nam
Sang-Yun Nam
   | Aug 31, 2017
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Article Category: Original Article
Published Online: Aug 31, 2017
Page range: 145 - 156
DOI: https://doi.org/10.5372/1905-7415.1102.546
Keywords
© 2017 Dan-Hee Kim, Sang-Yun Nam
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Effect of radish taffy on inflammatory activity of RAW264.7 cells in vitro. (Left panel) To examine the cytotoxicity of radish taffy, 10 μL of cell counting kit (CCK)-8 reagent in the final 1 h of incubation was added to cells exposed to 0–10 mg/ml of radish taffy and lipopolysaccharide, and then absorbance of the supernatant measured at 450 nm. (Right panel) To examine the effect of radish taffy on NO and cytokine production, cells were exposed to 0.01 or 0.1 mg/mL of radish taffy and 5 μM of PEITC for 20 h and then NO and cytokine levels in the supernatants were measured. Accumulated data from 2 separate experiments with similar results are shown and data are presented as means ± standard error of the mean. ∗∗P < 0.01 (n = 8).
Effect of radish taffy on inflammatory activity of RAW264.7 cells in vitro. (Left panel) To examine the cytotoxicity of radish taffy, 10 μL of cell counting kit (CCK)-8 reagent in the final 1 h of incubation was added to cells exposed to 0–10 mg/ml of radish taffy and lipopolysaccharide, and then absorbance of the supernatant measured at 450 nm. (Right panel) To examine the effect of radish taffy on NO and cytokine production, cells were exposed to 0.01 or 0.1 mg/mL of radish taffy and 5 μM of PEITC for 20 h and then NO and cytokine levels in the supernatants were measured. Accumulated data from 2 separate experiments with similar results are shown and data are presented as means ± standard error of the mean. ∗∗P < 0.01 (n = 8).

Figure 2

Dose–response effect of radish taffy (RT) on recruitment of leukocytes to airways in mice with induced pulmonary inflammation. Mice received 0.2–2 g/kg of RT (100 μL) orally or an equal volume of sterilized distilled water as a control for 10 days. One hour after the last administration of RT, pulmonary inflammation was induced by instillation of lipopolysaccharide (LPS 10 μg) and bronchoalveolar lavage fluid was analyzed 24 h later. Accumulated data from 2 separate experiments with similar results are shown. ∗∗P < 0.01 (n = 8).
Dose–response effect of radish taffy (RT) on recruitment of leukocytes to airways in mice with induced pulmonary inflammation. Mice received 0.2–2 g/kg of RT (100 μL) orally or an equal volume of sterilized distilled water as a control for 10 days. One hour after the last administration of RT, pulmonary inflammation was induced by instillation of lipopolysaccharide (LPS 10 μg) and bronchoalveolar lavage fluid was analyzed 24 h later. Accumulated data from 2 separate experiments with similar results are shown. ∗∗P < 0.01 (n = 8).

Figure 3

Effect of radish taffy (RT) on the cellularity in bronchoalveolar lavage fluid (BALF). (A) Total leukocyte numbers in BALF were counted using a hemacytometer (∗∗P < 0.01, n = 10) and (B) the numbers of polymorphonuclear leukocytes (PMN), and (C) mononuclear cells (MNC) were calculated with cytospin preparations stained with Diff-Quik. ∗∗P < 0.01 (n = 10).
Effect of radish taffy (RT) on the cellularity in bronchoalveolar lavage fluid (BALF). (A) Total leukocyte numbers in BALF were counted using a hemacytometer (∗∗P < 0.01, n = 10) and (B) the numbers of polymorphonuclear leukocytes (PMN), and (C) mononuclear cells (MNC) were calculated with cytospin preparations stained with Diff-Quik. ∗∗P < 0.01 (n = 10).

Figure 4

Effect of radish taffy (RT) on myeloperoxidase (MPO) expression in bronchoalveolar lavage fluid (BALF) and lung tissue. BALF and lung homogenates were prepared and MPO levels were determined using an ELISA. ∗∗P < 0.01 (n = 10).
Effect of radish taffy (RT) on myeloperoxidase (MPO) expression in bronchoalveolar lavage fluid (BALF) and lung tissue. BALF and lung homogenates were prepared and MPO levels were determined using an ELISA. ∗∗P < 0.01 (n = 10).

Figure 5

Effect of radish taffy (RT) on cytokine secretion. Levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL) 6, IL 10, and IL 12 were determined with ELISAs in bronchoalveolar lavage fluid (BALF) (left panel) and lung homogenates (right panel). ∗∗P < 0.01 (n = 14).
Effect of radish taffy (RT) on cytokine secretion. Levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL) 6, IL 10, and IL 12 were determined with ELISAs in bronchoalveolar lavage fluid (BALF) (left panel) and lung homogenates (right panel). ∗∗P < 0.01 (n = 14).

Figure 6

Effect of radish taffy (RT) on nuclear factor (NF)-κB activation. Total and phosphorylated NF-κB p65 (Ser536) levels were determined in lung homogenates using a PathScan sandwich ELISA kit (n = 4).
Effect of radish taffy (RT) on nuclear factor (NF)-κB activation. Total and phosphorylated NF-κB p65 (Ser536) levels were determined in lung homogenates using a PathScan sandwich ELISA kit (n = 4).

Figure 7

Total ion chromatograms of allicin (A), glycyrrhizin (B) and gingerol (C) for each standard (upper panels) and radish taffy sample (lower panels) using liquid chromatography-mass spectrometry (HPLC/MS) on a 1200 series HPLC system (Agilent Technologies) using a reversed phase column in the acquisition condition. Mass spectrometric detection was performed using Agilent 6410B instrument and ions were generated in positive ionization mode using electrospray ionization interface (intensity vertical axis). The fragmentor potential was 110 V and the interface heater 300 °C.
Total ion chromatograms of allicin (A), glycyrrhizin (B) and gingerol (C) for each standard (upper panels) and radish taffy sample (lower panels) using liquid chromatography-mass spectrometry (HPLC/MS) on a 1200 series HPLC system (Agilent Technologies) using a reversed phase column in the acquisition condition. Mass spectrometric detection was performed using Agilent 6410B instrument and ions were generated in positive ionization mode using electrospray ionization interface (intensity vertical axis). The fragmentor potential was 110 V and the interface heater 300 °C.

Figure 8

Total ion chromatogram of standard phenethyl isothiocyanate (PEITC) (A) and radish taffy sample (B) using of gas chromatography–mass spectrometry (GC/MS). Gas chromatography was performed on a DB-5MS fused-silica capillary column using a mass selective detector (Shimadzu GCMS-QP2010 Ultra) equipped with Labsolution software and National Institute of Standards and Technology spectra data.
Total ion chromatogram of standard phenethyl isothiocyanate (PEITC) (A) and radish taffy sample (B) using of gas chromatography–mass spectrometry (GC/MS). Gas chromatography was performed on a DB-5MS fused-silica capillary column using a mass selective detector (Shimadzu GCMS-QP2010 Ultra) equipped with Labsolution software and National Institute of Standards and Technology spectra data.

Quantification of 4 anti-inflammatory compounds in radish taffy by HPLC/MS

CompoundRetention timeSample peak area*

Sample was diluted to 40 mg/mL in distilled water and injection volume was 3 μL. ND = not detected.

Content in sample (μg/g)
Allicin4.583130.1
Glycyrrhizin6.287682,380
6-Gingerol6.5628862.0
Phenethyl isothiocyanate13.87NDND
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