J Korean Med Sci.  2012 Jul;27(7):777-783. 10.3346/jkms.2012.27.7.777.

Methionine Enhances the Contractile Activity of Human Colon Circular Smooth Muscle In Vitro

• Choe EK
• Moon JS
• Park KJ

• KMID: 2157923
• DOI: http://doi.org/10.3346/jkms.2012.27.7.777

Abstract

Effective drug to manage constipation has been unsatisfactory. We sought to determine whether methionine has effect on the human colon. Human colon tissues were obtained from the specimens of colon resection. Microelectrode recording was performed and contractile activity of muscle strips and the propagation of the contractions in the colon segment were measured. At 10 microM, methionine depolarized the resting membrane potential (RMP) of circular muscle (CM) cells. In the CM strip, methionine increased the amplitude and area under the curve (AUC) of contractions. In the whole segment of colon, methionine increased the amplitude and AUC of the high amplitude contractions in the CM. These effects on contraction were maximal at 10 microM and were not observed in longitudinal muscles in both the strip and the colon segment. Methionine reversed the effects of pretreatment with sodium nitroprusside, tetrodotoxin and Nw-oxide-L-arginine, resulting in depolarization of the RMP, and increased amplitude and AUC of contractions in the muscle strip. Methionine treatment affected the wave pattern of the colon segment by evoking small sized amplitude contractions superimposed on preexisting wave patterns. Our results indicate that a compound mimicking methionine may provide prokinetic functions in the human colon.

Keyword

Colon; Methionine; Gastrointestinal Motility; Humans

MeSH Terms

Area Under Curve
Arginine/pharmacology
Colon/drug effects/physiology
Humans
Membrane Potentials/drug effects
Methionine/*pharmacology
Microelectrodes
Muscle Contraction/*drug effects
Muscle, Smooth/drug effects/*physiology
Nitroprusside/pharmacology
Tetrodotoxin/pharmacology
Nitroprusside
Tetrodotoxin
Methionine
Arginine

Figure

• 

  Fig. 1 Effect of methionine on the membrane potential of human colon circular muscle. (A) Methionine at 10 µM concentration caused depolarization of the resting membrane potential. (B) Various concentrations of methionine (5, 10, 100, 500 µM) were used to treat circular muscle. 10 µM of methionine treatment resulted in maximal depolarization in the resting membrane potential of circular muscle. The figure shows the resting membrane potential according to the concentration of methionine.

• 

  Fig. 2 Effect of methionine on the membrane potential of human colon circular muscle in the presence of pretreated drugs. Pretreatment drug was perfused at 15 min intervals to 10 µM methionine. (A) Pretreating with 100 µM SNP hyperpolarized the RMP. 10 µM of methionine depolarized the RMP and overcame the effect of SNP. (B) Pretreating with 1 µM TTX depolarized the RMP. 10 µM methionine depolarized the RMP and had additional effect over TTX. (C) Pretreating with 100 µM NOLA depolarized the RMP. 10 µM methionine depolarized the RMP and had additional effect over NOLA. *The horizontal line is the reference to compare the RMP after drug treatment.

• 

  Fig. 3 Effect of methionine on the contraction of human colon CM strip. (A) After 10 µM methionine treatment on CM, amplitude and AUC for one minute increased. The black vertical line indicates the starting point of 10 µM methionine perfusion. (B) First KRB solution, then 1 µM TTX and 10 µM methionine were perfused at 45 min time intervals. TTX had a neurogenic blocking effect and inhibited the contraction in the CM strip. Note that after perfusing 10 µM methionine in the presence of TTX, the amplitude and area under the curve for one minute increased, indicating that 10 µM of methionine treatment had overcome the blocking effect of TTX and enhanced the contraction of the CM strip. The data in the open circle is magnified at the second row. (C) First KRB solution, then 100 µM NOLA and 10 µM methionine was perfused at 45 min time intervals. NOLA enhanced the contraction in the CM strip. Note that after perfusing 10 µM methionine, the amplitude and AUC increased further. 10 µM of methionine treatment had additional effect over NOLA.

• 

  Fig. 4 Effect of methionine on human colon segment tension. (A) Methionine at 10 µM concentration caused and increase in amplitude and AUC in the dominant waves of the circular muscle. (B) Various concentrations of methionine (10, 50, 100 µM) were used \to treat the circular muscle of the human colon segment. 10 µM methionine treatment resulted in a maximal increase in amplitude and AUC of the dominant waves in the circular muscle of the colon segment. The figure shows the amplitude according to the concentration of treated methionine. (C) 10 µM of methionine had no effect on longitudinal muscle motility.

• 

  Fig. 5 Effect of methionine on wave patterns of human colon segment circular muscle. In 3 samples, a non-dominant wave newly appeared after treating with 10 µM methionine. The black vertical line indicates the starting point of the 10 µM methionine perfusion.

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