Lab Anim Res.  2014 Mar;30(1):35-43.

Metabolomics approach to serum biomarker for loperamide-induced constipation in SD rats

• Kim JE
• Lee YJ
• Kwak MH
• Jun G
• Koh EK
• Song SH
• Seong JE
• Kim JW
• Kim KB
• Kim S
• Hwang DY

Abstract

Loperamide has long been known as an opioid-receptor agonist useful as a drug for treatment of diarrhea resulting from gastroenteritis or inflammatory bowel disease as well as to induce constipation. To determine and characterize putative biomarkers that can predict constipation induced by loperamide treatment, alteration of endogenous metabolites was measured in the serum of Sprague Dawley (SD) rats treated with loperamide for 3 days using 1H nuclear magnetic resonance (1H NMR) spectral data. The amounts and weights of stool and urine excretion were significantly lower in the loperamide-treated group than the No-treated group, while the thickness of the villus, crypt layer, and muscle layer was decreased in the transverse colon of the same group. The concentrations of aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and creatinine (Cr) were also slightly changed in the loperamide-treated group, although most of the serum components were maintained at a constant level. Furthermore, pattern recognition of endogenous metabolites showed completely separate clustering of the serum analysis parameters between the No-treated group and loperamide-treated group. Among 35 endogenous metabolites, four amino acids (alanine, glutamate, glutamine and glycine) and six endogenous metabolites (acetate, glucose, glycerol, lactate, succinate and taurine) were dramatically decreased in loperamide-treated SD rats. These results provide the first data pertaining to metabolic changes in SD rats with loperamide-induced constipation. Additionally, these findings correlate the changes in 10 metabolites with constipation.

Keyword

loperamide; constipation; metabolomics; serum; transverse colon

MeSH Terms

Amino Acids
Animals
Aspartate Aminotransferases
Biomarkers
Colon, Transverse
Constipation*
Creatinine
Diarrhea
Gastroenteritis
Glucose
Glutamic Acid
Glutamine
Glycerol
Inflammatory Bowel Diseases
L-Lactate Dehydrogenase
Lactic Acid
Loperamide
Magnetic Resonance Spectroscopy
Metabolomics*
Rats*
Succinic Acid
Weights and Measures
Amino Acids
Aspartate Aminotransferases
Creatinine
Glucose
Glutamic Acid
Glutamine
Glycerol
L-Lactate Dehydrogenase
Lactic Acid
Loperamide
Succinic Acid

Figure

• 

  Figure 1 Effects of loperamide treatment on histological parameters in the transverse colon of SD rats. (A) H&E-stained sections of transverse colons collected from No-treated rats (a and b) andloperamide-treated rats (c and d) were observed at two different magnifications using a light microscope. (B) The crypt number per specific area and diameter per crypt was measured with Leica Application Suite. Five to six rats per group were assayed in triplicate by H&E. Data represent the mean±SD from three replicates. *P<0.05 compared to the No-treated group.

• 

  Figure 2 Metabolomics pattern recognition using PCA (a) and OPLS-DA (b). (A) Global profiling of loperamide treatment in the serum samples. (B) Targeted profiling of loperamide treatment in the serum samples. (C) The VIP showed the major metabolites contributing to cluster separation.

• 

  Figure 3 Concentration of four amino acids after loperamide administration in SD rats. Data represent the mean±SD from three replicates. *P<0.05 compared to the No-treated group.

• 

  Figure 4 Concentration of six endogenous metabolites after loperamide administration in SD rats. Data represent the mean±SD from three replicates. *P<0.05 compared to the No-treated group.

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  Figure 5 Pathway related to endogenous metabolites that were changed after loperamide treatment. Bold arrows indicate the alteration of metabolites, while the box shows the altered metabolites.

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