J Rheum Dis.  2017 Aug;24(4):192-202. 10.4078/jrd.2017.24.4.192.

Metabolomics Approach to Explore the Effects of Rebamipide on Inflammatory Arthritis Using Ultra Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry

• Moon SJ
• Lee SH
• Jung BH
• Min JK

• KMID: 2389060
• DOI: http://doi.org/10.4078/jrd.2017.24.4.192

Abstract

OBJECTIVE
Rebampide is a gastroprotective agent used to treat gastritis. It possesses anti-inflammatory and anti-arthritis effects, but the mechanisms of these effects are not well understood. The objective of this study was to explore mechanisms underlying the therapeutic effects of rebamipide in inflammatory arthritis.
METHODS
Collagen-induced arthritis (CIA) was induced in DBA/1J mice. DBA/1J mice were immunized with chicken type II collagen, then treated intraperitoneally with rebamipide (10 mg/kg or 30 mg/kg) or vehicle (10% carboxymethylcellulose solution) alone. Seven weeks later, plasma samples were collected. Plasma metabolic profiles were analyzed using ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry-based metabolomics study and metabolite biomarkers were identified through multivariate data analysis.
RESULTS
Low dose rebamipide treatment reduced the clinical arthritis score compared with vehicle treatment, whereas high dose rebamipide in CIA aggravated arthritis severity. Based on multivariate analysis, 17 metabolites were identified. The plasma levels of metabolites associated with fatty acids and phospholipid metabolism were significantly lower with rebamipide treatment than with vehicle. The levels of 15-deoxy-Δ¹²,¹â´ prostaglandin J2 and thromboxane B3 decreased only in high dose-treated groups. Certain peptide molecules, including enterostatin (VPDPR) enterostatin and bradykinin dramatically increased in rebamipide-treated groups at both doses. Additionally, corticosterone increased in the low dose-treated group and decreased in the high dose-treated group.
CONCLUSION
Metabolomics analysis revealed the anti-inflammatory effects of rebamipide and suggested the potential of the drug repositioning in metabolism- and lipid-associated diseases.

Keyword

Rebamipide; Arthritis; Metabolomics; Anti-oxidant

MeSH Terms

Animals
Arthritis*
Arthritis, Experimental
Biomarkers
Bradykinin
Carboxymethylcellulose Sodium
Chickens
Collagen Type II
Corticosterone
Drug Repositioning
Fatty Acids
Gastritis
Mass Spectrometry*
Metabolism
Metabolome
Metabolomics*
Mice
Multivariate Analysis
Plasma
Statistics as Topic
Therapeutic Uses
Biomarkers
Bradykinin
Carboxymethylcellulose Sodium
Collagen Type II
Corticosterone
Fatty Acids
Therapeutic Uses

Figure

• 

  Figure 1. Treatment with rebamipide (10 mg/kg) suppresses inflammatory arthritis in mice with collagen-induced arthritis (CIA). CIA was induced in DBA/1J mice by immunization with type II collagen (CII) in adjuvant. Changes in arthritis score in rebamipide-treated mice compared with vehicle-treated mice. Rebamipide dissolved in 10% carboxymethylcellulose solution (vehicle) was given intraperitoneally to 2 different groups (each receiving 10 or 30 mg/kg; n=10 mice per group) daily for 4 weeks, starting after booster immunization. A third group (n=10) received vehicle alone. WT: wild-type, D: day.

• 

  Figure 2. (A) Principal component analysis (PCA) score plots and (B) partial least square-discriminant analysis (PLS-DA) score plots based on plasma metabolic profiling. (a) Positive ionization mode. (b) Negative ionization mode. In PCA, the score plot was obtained with the two PCs presenting 47.2% (PC1) and 14.5% (PC2) variance in positive ionization mode and that was with three PCs presenting 23.3% (PC1), 18.7% (PC2), and 13.2% (PC3) in negative ionization mode. In PLS-DA, the score plot was obtained with the two PCs presenting 43.2% (PC1) and 30.5% (PC2) variance in positive ionization mode and that was with three PCs presenting 53.7% (PC1), 38.6% (PC2), and 23.3% (PC3) in negative ionization mode.

• 

  Figure 3. Comparison of metabolites with significant changes involved in (A) peptide metabolism, (B) fatty acid metabolism, (C) phospholipid metabolism, (D) acylcarnitine (β-oxidation) metabolism, (E) prostaglandin metabolism, and (F) corticosteroid hormone metabolism. *p＜0.05, **p＜0.01, and ***p＜0.001 compared with vehicle control. HpODE: hydroxyoctadecadienoic acid, LysoPC: lysophosphatidylcholine, PGJ2: prostaglandin J2.

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