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Effect of Obstructive Jaundice and Nitric Oxide on the Profiles of Intestinal Bacterial Flora in Wild and iNOS Mice.

Hong JY, F Sato E, Nishikawa T, Hiramoto K, Inoue M - J Clin Biochem Nutr (2009)

Bottom Line: The amounts of bacterial DNA detected in fecal samples from both animal groups pretreated with antibiotics were extremely low as compared with untreated groups.We found that the profiles of enteric bacteria changed markedly after BDL.Among enteric bacteria examined, Lactobacillus murinus was found to increase markedly after BDL in rectum of both animal groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Pathology, Osaka City University Medical School, 1-4-3 Asahimachi, Abeno, Osaka 545-8585, Japan.

ABSTRACT
We previously reported that the plasma level of endotoxin and colonic expression of IgA in the mouse increased with obstructive jaundice induced by bile duct ligation (BDL). To elucidate the mechanism of the BDL-induced increase, we analyzed the effect of BDL on intestinal flora in wild type and inducible nitric oxide synthase (iNOS)-deficient mice (iNOS(-/-)) using the terminal restriction fragment length polymorphism analysis (T-RFLP) and 16S rDNA clone libraries. The amounts of bacterial DNA detected in fecal samples from both animal groups pretreated with antibiotics were extremely low as compared with untreated groups. We found that the profiles of enteric bacteria changed markedly after BDL. The bacterial composition is significantly different between not only wild type and iNOS(-/-) mice but also those before and after BDL, respectively. Among enteric bacteria examined, Lactobacillus murinus was found to increase markedly after BDL in rectum of both animal groups. However, Escherichia coli markedly increased after BDL in the iNOS(-/-) mice. These findings suggest that profiles of enteric flora change markedly in animals during obstructive jaundice by some mechanism that is affected by bile constituents and iNOS-derived NO.

No MeSH data available.


Related in: MedlinePlus

T-RFLP analysis of intestinal microbiota before and after BDL. Data shows T-RFLP patterns of 16S rDNA from mouse feces (day 0~7) digested with HhaI. 16S rDNA were amplified using universal primers 27F and 1492R. The minimum and maximum values of the ordinate of each T-RFLP pattern are 0 to 800 fluorescence intensity (arbitary units) for HhaI. Fecal samples from the rectum of (A) wild and (B) iNOS−/− mice, and the cerum of (C) wild and (D) iNOS−/− mice
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Figure 2: T-RFLP analysis of intestinal microbiota before and after BDL. Data shows T-RFLP patterns of 16S rDNA from mouse feces (day 0~7) digested with HhaI. 16S rDNA were amplified using universal primers 27F and 1492R. The minimum and maximum values of the ordinate of each T-RFLP pattern are 0 to 800 fluorescence intensity (arbitary units) for HhaI. Fecal samples from the rectum of (A) wild and (B) iNOS−/− mice, and the cerum of (C) wild and (D) iNOS−/− mice

Mentions: Although bacterial DNA was easily detected in fecal samples from wild and iNOS−/− mice, it was practically difficult to detect DNA from both animal groups pretreated with antibiotics (Fig. 1). TRFLP patterns of the wild and iNOS−/− mice were compared using two different restriction enzymes before and after BDL (Fig. 2 and 3). The profiles of bacterial community in the feces from wild and iNOS−/− mice changed markedly after BDL. The profiles of bacterial distribution in the feces from the rectum and the cecum also differed significantly after BDL. After digestion with HhaI, terminal restriction fragments (T-RFs) at bp 249 and 370, and bp 370 were detected in the feces from rectum and cecum of wild type mice after BDL. In contrast, HhaI digestion revealed T-RFs at bp 233, 249, and 370, and bp 197 and 208 in the feces from the rectum and cecum of iNOS−/− mice that had been subjected to BDL (Fig. 2). MspI digestion revealed T-RFs at bp 491, 492 and 565, and bp 491 in the feces from the rectum and cecum of BDL-treated wild type mice. MspI digestion revealed T-RFs at bp 492, 568 and 583, and bp 223 and 269 in the fecal sample from the rectum and cecum of BDL-treated iNOS−/− mice (Fig. 3).


Effect of Obstructive Jaundice and Nitric Oxide on the Profiles of Intestinal Bacterial Flora in Wild and iNOS Mice.

Hong JY, F Sato E, Nishikawa T, Hiramoto K, Inoue M - J Clin Biochem Nutr (2009)

T-RFLP analysis of intestinal microbiota before and after BDL. Data shows T-RFLP patterns of 16S rDNA from mouse feces (day 0~7) digested with HhaI. 16S rDNA were amplified using universal primers 27F and 1492R. The minimum and maximum values of the ordinate of each T-RFLP pattern are 0 to 800 fluorescence intensity (arbitary units) for HhaI. Fecal samples from the rectum of (A) wild and (B) iNOS−/− mice, and the cerum of (C) wild and (D) iNOS−/− mice
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654472&req=5

Figure 2: T-RFLP analysis of intestinal microbiota before and after BDL. Data shows T-RFLP patterns of 16S rDNA from mouse feces (day 0~7) digested with HhaI. 16S rDNA were amplified using universal primers 27F and 1492R. The minimum and maximum values of the ordinate of each T-RFLP pattern are 0 to 800 fluorescence intensity (arbitary units) for HhaI. Fecal samples from the rectum of (A) wild and (B) iNOS−/− mice, and the cerum of (C) wild and (D) iNOS−/− mice
Mentions: Although bacterial DNA was easily detected in fecal samples from wild and iNOS−/− mice, it was practically difficult to detect DNA from both animal groups pretreated with antibiotics (Fig. 1). TRFLP patterns of the wild and iNOS−/− mice were compared using two different restriction enzymes before and after BDL (Fig. 2 and 3). The profiles of bacterial community in the feces from wild and iNOS−/− mice changed markedly after BDL. The profiles of bacterial distribution in the feces from the rectum and the cecum also differed significantly after BDL. After digestion with HhaI, terminal restriction fragments (T-RFs) at bp 249 and 370, and bp 370 were detected in the feces from rectum and cecum of wild type mice after BDL. In contrast, HhaI digestion revealed T-RFs at bp 233, 249, and 370, and bp 197 and 208 in the feces from the rectum and cecum of iNOS−/− mice that had been subjected to BDL (Fig. 2). MspI digestion revealed T-RFs at bp 491, 492 and 565, and bp 491 in the feces from the rectum and cecum of BDL-treated wild type mice. MspI digestion revealed T-RFs at bp 492, 568 and 583, and bp 223 and 269 in the fecal sample from the rectum and cecum of BDL-treated iNOS−/− mice (Fig. 3).

Bottom Line: The amounts of bacterial DNA detected in fecal samples from both animal groups pretreated with antibiotics were extremely low as compared with untreated groups.We found that the profiles of enteric bacteria changed markedly after BDL.Among enteric bacteria examined, Lactobacillus murinus was found to increase markedly after BDL in rectum of both animal groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Pathology, Osaka City University Medical School, 1-4-3 Asahimachi, Abeno, Osaka 545-8585, Japan.

ABSTRACT
We previously reported that the plasma level of endotoxin and colonic expression of IgA in the mouse increased with obstructive jaundice induced by bile duct ligation (BDL). To elucidate the mechanism of the BDL-induced increase, we analyzed the effect of BDL on intestinal flora in wild type and inducible nitric oxide synthase (iNOS)-deficient mice (iNOS(-/-)) using the terminal restriction fragment length polymorphism analysis (T-RFLP) and 16S rDNA clone libraries. The amounts of bacterial DNA detected in fecal samples from both animal groups pretreated with antibiotics were extremely low as compared with untreated groups. We found that the profiles of enteric bacteria changed markedly after BDL. The bacterial composition is significantly different between not only wild type and iNOS(-/-) mice but also those before and after BDL, respectively. Among enteric bacteria examined, Lactobacillus murinus was found to increase markedly after BDL in rectum of both animal groups. However, Escherichia coli markedly increased after BDL in the iNOS(-/-) mice. These findings suggest that profiles of enteric flora change markedly in animals during obstructive jaundice by some mechanism that is affected by bile constituents and iNOS-derived NO.

No MeSH data available.


Related in: MedlinePlus