Eubacterium limosum ameliorates experimental colitis and metabolite of microbe attenuates colonic inflammatory action with increase of mucosal integrity - PubMed (original) (raw)

Eubacterium limosum ameliorates experimental colitis and metabolite of microbe attenuates colonic inflammatory action with increase of mucosal integrity

Osamu Kanauchi et al. World J Gastroenterol. 2006.

Abstract

Aim: To examine the effect of Eubacterium limosum (E. limosum) on colonic epithelial cell line in vitro, and to evaluate the effect of E. limosum on experimental colitis.

Methods: E. limosum was inoculated anaerobically and its metabolites were obtained. The growth stimulatory effect of the E. limosum metabolites on T84 cells was evaluated by SUDH activity, and the anti-inflammatory effect by IL-6 production. The change in mRNA of toll like receptor 4 (TLR4) was evaluated by real time PCR. Colitis was induced by feeding BALB/C mice with 2.0% dextran sodium sulfate. These mice received either 5% lyophilized E. limosum (n = 7) or control diet (n = 7). Seven days after colitis induction, clinical and histological scores, colon length, and cecal organic acid levels were determined.

Results: The E. limosum produced butyrate, acetate, propionate, and lactate at 0.25, 1.0, 0.025 and 0.07 mmol/L, respectively in medium. At this concentration, each acid had no growth stimulating activity on T84 cells; however, when these acids were mixed together at the above levels, it showed significantly high activity than control. Except for lactate, these acids significantly attenuated IL-6 production at just 0.1 mmol/L. In addition, under TNF-alpha stimulation, butyrate attenuated the production of TLR4 mRNA. The treatment with E. limosum significantly attenuated clinical and histological scores of colitis with an increase of cecal butyrate levels, compared with the control group.

Conclusion: E. limosum can ameliorate experimental colonic inflammation. In part, the metabolite of E. limosum, butyrate, increases mucosal integrity and shows anti-inflammatory action modulation of mucosal defense system via TLR4.

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Figures

Figure 5

Figure 5

Disease activity index after initiation of dextran sulfate sodium-induced colitis. Control indicates the control diet (cellulose diet) group. E.limisum indicates the Eubacterium limosum diet group.Data is expressed as means ± SD (n = 7). a_P_ < 0.05 vs control. Disease activity index was determined by scoring changes in body weight, hemoccult positivity, and stool consistency.

Figure 1

Figure 1

Effect of organic acids (A) and the cocktail mixture of organic acids (B) on SUDH activity in T84 cells. Cells were incubated with various concentrations of fatty acids for 24 hours. The data is expressed as means ± SD (n = 4). a_P_ < 0.05 vs control. 1The cocktail was an organic acid mixture adjusted to that of 10 % E. limosum supernatant (butyrate [0.25 mmol/L], acetate [1 mmol/L], propionate (0.025 mmol/L), and lactate [0.07 mmol/L]).

Figure 2

Figure 2

Effect of organic acids on secretion of IL-6 by TNF-alpha stimulation in T84 cells. Cells were incubated with TNF-alpha (50 ng/mL) and various concentrations of organic acids for 24 hours, then supernatant was applied for determination of IL-6 production by ELISA. Data is expressed as means ± SD (n = 3). a_P_ < 0.05 vs control.

Figure 3

Figure 3

Inhibition of response to TNF -alpha (25 ng/mL) induced the mRNA level of toll-like receptor 4 (TLR4) in T84 by exposure to butyrate. Cells were treated with 0 to 2.0 mmol/L sodium butyrate for 6 hr prior to extraction of RNA and real time PCR (RT-PCR). RT-PCR was performed with specific primers and probs for indicated receptor components and housekeeping gene beta-actin. All data were normalized to the expression of beta-actin, and presented as % value of control (without TNF-alpha treatment). Data is expressed as means ± SD.

Figure 4

Figure 4

Changes in body weight after initiation of dextran sulfate sodium-induced colitis. control indicates the control diet (cellulose diet) group. E.limisum indicates the Eubacterium limosum diet group. Data is expressed as means ± SD (n = 7). a_P_<0.05 vs control.

Figure 6

Figure 6

Light histological section of colonic mucosa (A) and changes in colon length and colonic mucosal damage score (B) of dextran sulfate sodium-induced colitis control and E.limisum diet groups. (magnification X 50). Control indicates the control diet (cellulose diet) group. E.limisum indicates the Eubacterium limosum diet group. Data is expressed as means ± SD (n = 7). a_P_ < 0.05 vs control.

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