Abstract
Reflux esophagitis (RE) is a gastrointestinal disease caused by the reflux of gastric acid and stomach contents, and it leads to esophageal damage. Therefore, it is necessary to study the improvement of esophageal damage on a RE-induced model. The present study was accomplished to demonstrate the protective effects of a dichloromethane fraction of Geranium koreanum (DGK) plant on esophageal damage in an acute RE rat model. First, we examined the potential of anti-inflammatory effects of various fractions measured by cell cytotoxicity, morphological changes and nitric oxide (NO) production on lipopolysaccharide (LPS)-induced Raw 264.7 macrophage cells. Then, to evaluate the protective effects on RE, rats were partitioned into the following groups: normal control, RE-induced control and RE rats pre-treated with DGK 100 and 200 mg/kg body weight. The esophageal mucosal ulcer ratio was measured by the Image J program and histological changes were examined using a hematoxylin and eosin staining of the esophageal mucosa. The expression of pro-inflammatory proteins, cytokines and tight junction proteins involved in the esophageal mucosal damage were investigated using Western blotting and an enzyme-linked immunosorbent assay (ELISA) kit with esophagus tissue. DGK chemical profile and phenolic contents were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). The results showed that DGK exhibited anti-inflammatory effects against LPS-stimulated cells by significantly inhibiting NO production. Additionally, the results in vivo showed that improvement effects of DGK on esophageal mucosal damage. The expression of inflammatory proteins involved in nuclear factor κB (NF-κB) signaling pathways and tight junction protein (claudin-4 and -5) were significantly decreased in esophageal mucosa. We found the potential of DGK as source of replacement therapy products for inflammatory and RE disease.
Highlights
Gastroesophageal reflux disease (GERD) is usually induced by the reflux of gastric acid or stomach contents into the esophagus due to a defection of the lower esophageal sphincter and it related to unphysiological stress, excessive drinking, smoking, sleeping position, crapulence, and Westernization of the lifestyle and diet
To identified the cytotoxicity of HEX, DICHO, ethyl acetate (EA) and BUT fraction of Geranium koreanum, we analyzed the cell viability on Raw 264.7 macrophage cells treated with LPS (1 μg/mL) for 24 h using a cytotoxicity assay kit
We found that dichloromethane fraction of Geranium koreanum (DGK) has anti-inflammatory effects via inhibition of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) protein expression in the LPS-induced macrophage
Summary
Gastroesophageal reflux disease (GERD) is usually induced by the reflux of gastric acid or stomach contents into the esophagus due to a defection of the lower esophageal sphincter and it related to unphysiological stress, excessive drinking, smoking, sleeping position, crapulence, and Westernization of the lifestyle and diet. Reflux esophagitis (RE) causes symptoms such as heartburn, nausea, chronic cough, pharyngeal pain and asthma and may lead to complication of Barrett esophagus, esophagus stenosis and esophageal cancer when the disease continues for a long time [1,2,3,4]. Proton pump inhibitors (PPIs) and histamine receptor antagonist forms of treatment have been widely used for rapid relief of symptoms and mucosal injury in patients with reflux disease, but these drugs cause many complications and are resistant to long-term use [5,6]. It was reported that protective effects of herbal extracts on tissue damage are attributed to cellular defense mechanisms such as an inflammation and oxidative stress responses [7,8,9]
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