Korean J Gastroenterol.  2009 Jul;54(1):20-27. 10.4166/kjg.2009.54.1.20.

Therapeutic Effect of Allogenic Bone Marrow Transplantation in Acute TNBS-induced Colitis

• Maeng LS
• Chang ED
• Chae HS
• Kim JS
• Min JY
• Sohn HS
• Rho SY
• Kim HK
• Cho YS
• Choi KY
• Lee HK

• KMID: 1792762
• DOI: http://doi.org/10.4166/kjg.2009.54.1.20

Abstract

BACKGROUND/AIMS
Bone marrow-derived cells (BMDC) contribute to tissue maintenance under many kinds of pathologic conditions. We carried out a study to see how BMDC play a role in the treatment of experimental murine colitis. METHODS: We divided the animals into 3 groups and treated them with 50% ethanol (control group), 2,4,6-trinitrobenzene sulfinic acid colitis (TNBS group), and TNBS+bone marrow transplant (BMT group). To induce colitis, TNBS (5.0 mg/mouse) dissolved in 50% ethanol was injected into anus weekly for two weeks. Bone marrow transplantations were performed using bone marrow of male transgenic mouse (donor) with green fluoresence protein (GFP) into female wild type mouse (recipient) three weeks before TNBS instillation. All animals were sacrificed, and colons were extracted one week after the last TNBS instillation. We measured microscopic scores of mucosal injury and investigated the GFP expression for bone marrow engraftment. The immunostaining of vimentin and alpha-smooth muscle actin (alpha-SMA) for myofibroblasts was performed. RESULTS: The score of mucosal injury in the TNBS group was much more severe than those in control, and reduced significantly by BMT (p<0.05). GFP-positive cells were almost deposited in pericryptal niche of BMT group but not at all in both control and TNBS group. Most of myofibroblasts stained with both vimentin and SMA also infiltrated into pericryptal niche. But, the number of myofibroblasts stained with vimentin and SMA in both control and TNBS group was smaller than that in BMT group. CONCLUSIONS: BMDC deposited on pericryptal niche might have a significant role in repairing acute experimental murine colitis.

Keyword

Bone marrow transplantation; TNBS-induced colitis

MeSH Terms

Actins/metabolism
Acute Disease
Animals
*Bone Marrow Transplantation
Colitis/chemically induced/pathology/*surgery
Female
Fibroblasts/cytology
Intestinal Mucosa/cytology
Male
Mice
Mice, Inbred C57BL
Mice, Transgenic
Transplantation, Homologous
Trinitrobenzenesulfonic Acid/*toxicity
Vimentin/metabolism

Figure

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  Fig. 1. Serial body weight changes of TNBS and BMT group.

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  Fig. 2. Microscopic findings show that the mucosal injury of TNBS colitis (score 3) is more severe than that of control (score 0) or BMT group (score 1). (A) 50% E-OH group, (B) TNBS group, (C) BMT group (H&E, ×200).

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  Fig. 3. Comparison of the mean ulcer score among three groups by H&E staining. It showed that the mean ulcer score of TNBS group is higher than those of control and BMT group.

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  Fig. 4. GFP positive cells providing evidence for successful bone marrow transplantation were found only in BMT group but not in both control and TNBS group. (A) 50% E-OH group, (B) TNBS group, (C, D) BMT group (IHC, ×200).

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  Fig. 5. Myofibroblasts stained with vimentin were mainly deposited in pericryptial area, and the number of positive cells in BMT group was higher than that in control or TNBS group. (A) 50% E-OH group, (B) TNBS group, (C) BMT group (IHC, ×200).

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  Fig. 6. α-SMA positive cells for detection of myofibroblasts are also located in pericryptal area and the number of positive cells is much higher in BMT group than in control or TNBS group. (A) 50% E-OH group, (B) TNBS group, (C, D) BMT group (Immunofluorescence staining, ×400).

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