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Anti-Inflammation and Protective Effects of Anethum graveolens L. (Dill Seeds) on Esophageal Mucosa Damages in Reflux Esophagitis-Induced Rats

Nam, Hyeon-Hwa, Nan, Li, Choo, Byung-Kil·2021·Foods
DOI 10.3390/foods10102500PMC PMC8535990
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Abstract

Anethum graveolens L. (dill seeds) are important medicinal and functional foods in Europe and central and south Asia, often used as a seasoning in daily diets. Anethum graveolens L. seeds (AGS) are used to treat indigestion and have shown physiological activities such as those against hypoglycemia and gastroesophageal disease. This study explored the protective effects of AGS extract on mucosal damages and inflammation in reflux esophagitis rats. AGS inhibited cellular inflammation including NO production and the expression of inflammatory proteins (iNOS and COX2 etc.), cytokines (IL-1β and TNF-α) and nuclear transfer factor related to NF-κB signaling caused by LPS stimulation in vitro. Furthermore, reflux esophagitis-induced rats were used to observe the anti-inflammatory effect of AGS. Tissue staining and inflammation-related protein expression of rats with acute reflux esophagitis indicated that AGS improved this inflammatory response, such as COX-2 and TNF-α in mucosa. In conclusion, AGS have good physiological activity and the possibility of being used as a medicinal food and a functional resource for the prevention and therapy of gastroesophageal diseases.

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Crossref · 10.3390/foods10102500

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Figures

Figure 1 Effects of Anethum graveolens L. seeds (AGS) in cell cytotoxicity and nitrite oxide (NO) production. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 18 h. Effect of AGS on morphological transformation (A), cell viability (B) and production of NO (C) in LPS-induced RAW264.7 cells. Scale bar was 200 μm. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).
Figure 1 Effects of Anethum graveolens L. seeds (AGS) in cell cytotoxicity and nitrite oxide (NO) production. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 18 h. Effect of AGS on morphological transformation (A), cell viability (B) and production of NO (C) in LPS-induced RAW264.7 cells. Scale bar was 200 μm. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).CC BY · PubMed Central
Figure 2 Inhibition effect of Anethum graveolens L. seeds (AGS) on cytokines production. Inhibition effect of AGS on production of TNF-α (A) and IL-1β (B) in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).
Figure 2 Inhibition effect of Anethum graveolens L. seeds (AGS) on cytokines production. Inhibition effect of AGS on production of TNF-α (A) and IL-1β (B) in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).CC BY · PubMed Central
Figure 3 Inhibition effect of Anethum graveolens L. seeds (AGS) on expression of inflammatory proteins. Inhibition effect of AGS on expression level (A) of iNOS (B), COX-2 (C), p-NF-κB (D) and p-IκBα (E) proteins in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 1h and 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05 and *** p < 0.001).
Figure 3 Inhibition effect of Anethum graveolens L. seeds (AGS) on expression of inflammatory proteins. Inhibition effect of AGS on expression level (A) of iNOS (B), COX-2 (C), p-NF-κB (D) and p-IκBα (E) proteins in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 1h and 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05 and *** p < 0.001).CC BY · PubMed Central
Figure 4 Inhibition the NF-kB Translocation by AGS. Inhibition effect of AGS nuclear transfer of NF-κB in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 1 h.
Figure 4 Inhibition the NF-kB Translocation by AGS. Inhibition effect of AGS nuclear transfer of NF-κB in LPS-induced RAW264.7 cells. Cells treated with the AGS (12.5, 25, 50 μg/mL) and LPS (1 μg/mL) for 1 h.CC BY · PubMed Central
Figure 5 Observation of the esophageal mucosa damages. The sketch map of surgery of the reflux esophagitis rat model (A), representative microscopic image of the esophageal mucosa damage (B), ratio of esophageal damage (C), pH and volume of gastric contents (D), of the esophageal mucosa. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).
Figure 5 Observation of the esophageal mucosa damages. The sketch map of surgery of the reflux esophagitis rat model (A), representative microscopic image of the esophageal mucosa damage (B), ratio of esophageal damage (C), pH and volume of gastric contents (D), of the esophageal mucosa. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (*** p < 0.001).CC BY · PubMed Central
Figure 6 The histological observation on esophageal mucosa. Scale bar was 200 μm.
Figure 6 The histological observation on esophageal mucosa. Scale bar was 200 μm.CC BY · PubMed Central
Figure 7 Expression of inflammatory proteins in esophagus. The expression of COX-2 (A) and TNF-α (B) in esophageal tissue. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05 and ** p < 0.01).
Figure 7 Expression of inflammatory proteins in esophagus. The expression of COX-2 (A) and TNF-α (B) in esophageal tissue. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05 and ** p < 0.01).CC BY · PubMed Central
Figure 8 Phytochemistry and functional properties of Anethum graveolens seeds (AGS). Chromatogram of 70% ethanolic extract of AGS (A); chlorogenic acid at 10.2 min, linoleic acid at 35.1 min, and elaidic acid at 39.5 min using HPLC-ELSD and Luna C8 (2) column. Anti-inflammatory effects of chlorogenic acid (CA), linoleic acid (LA), and elaidic acid (EA) in LPS-induced RAW 264.7 cells (B). Cells treated with the chlorogenic acid and elaidic acid (31.3, 62.5, 125 μM), linoleic acid (3.13, 6.25, 12.5 μM), and LPS (1 μg/mL) for 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05, ** p < 0.01 and *** p < 0.001).
Figure 8 Phytochemistry and functional properties of Anethum graveolens seeds (AGS). Chromatogram of 70% ethanolic extract of AGS (A); chlorogenic acid at 10.2 min, linoleic acid at 35.1 min, and elaidic acid at 39.5 min using HPLC-ELSD and Luna C8 (2) column. Anti-inflammatory effects of chlorogenic acid (CA), linoleic acid (LA), and elaidic acid (EA) in LPS-induced RAW 264.7 cells (B). Cells treated with the chlorogenic acid and elaidic acid (31.3, 62.5, 125 μM), linoleic acid (3.13, 6.25, 12.5 μM), and LPS (1 μg/mL) for 18h. Data were expressed as mean ± SD of duplicate experiment. Statistical analysis was performed for LPS compared with normal cells (### p < 0.001) and each sample compared with LPS-induced cells (* p < 0.05, ** p < 0.01 and *** p < 0.001).CC BY · PubMed Central

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Citations

Cites

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Metadata

Authors
Nam, H.-H., Nan, L., & Choo, B.-K.
Venue
Foods
Publisher
MDPI AG
Published
2021-10-18
Volume
10
Issue
10
Pages
2500
ISSN
2304-8158
DOI
10.3390/foods10102500
PMC
PMC8535990
Cited by
22
References
1
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
  • Rural Development Administration (PJ0142272020)
  • Jeonbuk National University (2020)

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