Lab Anim Res.  2016 Dec;32(4):231-240. 10.5625/lar.2016.32.4.231.

Characterization the response of Korl:ICR mice to loperamide induced constipation

• Kim JE
• Yun WB
• Sung JE
• Lee HA
• Choi JY
• Choi YS
• Jung YS
• Kim KS
• Hwang DY

• KMID: 2362915
• DOI: http://doi.org/10.5625/lar.2016.32.4.231

Abstract

Animal models of constipation induced with drugs and diet have been widely employed to investigate therapeutic effects and the action mechanism of drugs against this disease. ICR mice were selected to produce this disease model through oral administration of loperamide (Lop), even though SD rats are commonly utilized in studies of constipation. To compare the responses of ICR mice obtained from three different sources to constipation inducers, alterations in stool number, histopathological structure, mucin secretion and opioid-receptor downstream signaling pathway were measured in Korl:ICR (Korea FDA source), A:ICR (USA source) and B:ICR (Japan source) injected with low and high concentrations of Lop (LoLop and HiLop). The number, weight and moisture content of stools decreased significantly in the Lop treated group of all ICR relative to the Vehicle treated group. Additionally, decreased mucosa layer thickness, muscle thickness, and mucin secretion were observed in the transverse colon of Lop treated ICR mice, while a similar number of goblet cells and crypt of lieberkuhn were detected in the same group. Furthermore, a similar change in the level of Gα expression and PKC phosphorylation was detected in the Lop treated group relative to the vehicle treated group, while some differences in the change pattern were observed in the B:ICR group. Therefore, these results of the present study provide strong additional evidence that Korl:ICR, A:ICR and B:ICR derived from different sources have a similar overall response to constipation induced by Lop injection, although there were a few differences in the magnitude of their responses.

Keyword

Korl:ICR; constipation; loperamide; excretion parameters; mucin secretion

MeSH Terms

Administration, Oral
Animals
Colon, Transverse
Constipation*
Diet
Goblet Cells
Loperamide*
Mice*
Mice, Inbred ICR
Models, Animal
Mucins
Mucous Membrane
Phosphorylation
Rats
Therapeutic Uses
Loperamide
Mucins
Therapeutic Uses

Figure

• 

  Figure 1 Strategy for induction of chronic constipation using Lop. After the first injection of Lop (4 mg/kg) for 4 days, ICR mice were allowed to rest for 3 days. Two different concentrations (4 and 8 mg/kg) of Lop were then subcutaneously injected into three groups of ICR mice to produce constipated mice.

• 

  Figure 2 Alteration of the number and morphology of stools in three ICR groups after Lop injection. (A) The number of stools were observed during two Lop injection phases and one stationary phase in Vehicle, LoLop and HiLop treated mice. Numbers were counted three times and data represent the means±SD of three replicates. *, P<0.05 compared to the Korl:ICR group. (B) Following the final treatment, total stools were collected from metabolic cages and their morphology was observed.

• 

  Figure 3 Alteration of histopathological structure in Lop-induced constipated ICR mice. H&E stained sections of transverse colons rats from the Vehicle treated group, LoLop treated group or HiLop treated group were observed at 200× magnification using a light microscope. The length of the mucosa layer and muscle thickness in each tissue were measured using Leica Application Suite. Data represent the means±SD of three replicates. *, P<0.05 compared to the Korl:ICR group.

• 

  Figure 4 Mucin secretion in the transverse colon of Lop-induced constipated ICR mice. Mucin secreted from crypt layer cells was stained with alcian blue at pH 2.5 and their morphology were observed at 200× magnification. Five to six rats per group were assayed in triplicate by alcian blue staining.

• 

  Figure 5 Alteration of the level of Gα expression and PKC phosphorylation. The Gα expression and PKC phosphorylation within opioid-receptor downstream signaling pathway was measured by Western blotting using HRP-labeled anti-rabbit IgG antibody. After the intensity of each band was determined using an imaging densitometer, the relative levels of protein were calculated based on the intensity of actin protein. Data represent the means±SD of three replicates. *, P<0.05 compared to the Korl:ICR group.

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