Moxibustion inhibits interleukin-12 and tumor necrosis factor alpha and modulates intestinal flora in rat with ulcerative colitis

AIM: To investigate the effect of moxibustion on intestinal flora and release of interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) from the colon in rat with ulcerative colitis (UC). METHODS: A rat model of UC was established by local stimulation of the intestine with supernatant from colonic contents harvested from human UC patients. A total of 40 male Sprague-Dawley rats were randomly divided into the following groups: normal (sham), model (UC), herb-partition moxibustion (HPM-treated), and positive control sulfasalazine (SA-treated). Rats treated with HPM received HPM at acupuncture points ST25 and RN6, once a day for 15 min, for a total of 8 d. Rats in the SA group were perfused with SA twice a day for 8 d. The colonic histopathology was observed by hematoxylin-eosin. The levels of intestinal flora, including Bifidobacterium, Lactobacillus, Escherichia coli (E. coli), and Bacteroides fragilis (B. fragilis), were tested by real-time quantitative polymerase chain reaction to detect bacterial 16S rRNA/DNA in order to determine DNA copy numbers of each specific species. Immunohistochemical assays were used to observe the expression of TNF-alpha and IL-12 in the rat colons. RESULTS: HPM treatment inhibited immunopathology in colonic tissues of UC rats; the general morphological score and the immunopathological score were significantly decreased in the HPM and SA groups compared with the model group [3.5 (2.0-4.0), 3.0 (1.5-3.5) vs 6.0 (5.5-7.0), P < 0.05 for the general morphological score, and 3.00 (2.00-3.50), 3.00 (2.50-3.50) vs 5.00 (4.50-5.50), P < 0.01. for the immunopathological score]. As measured by DNA copy number, we found that Bifidobacterium and Lactobacillus, which are associated with a healthy colon, were significantly higher in the HPM and SA groups than in the model group (1.395 +/- 1.339, 1.461 +/- 1.152 vs 0.045 +/- 0.036, P < 0.01 for Bifidobacterium, and 0.395 +/- 0.325, 0.851 +/- 0.651 vs 0.0015 +/- 0.0014, P < 0.01 for Lactobacillus). On the other hand, E coli and B. fragilis, which are associated with an inflamed colon, were significantly lower in the HPM and SA groups than in the model group (0.244 +/- 0.107, 0.628 +/- 0.257 vs 1.691 +/- 0.683, P < 0.01 for E. coli, and 0.351 +/- 0.181, 0.416 +/- 0.329 vs 1.285 +/- 1.039, P < 0.01 for B. fragills). The expression of TNF-alpha and IL-12 was decreased after HPM and SA treatment as compared to UC model alone (4970.81 +/- 959.78, 6635.45 +/- 1135.16 vs 12333.81 +/- 680.79, P < 0.01 for TNF-alpha, and 5528.75 +/- 1245.72, 7477.38 +/- 1259.16 vs 12550.29 +/- 1973.30, P < 0.01 for IL-12). CONCLUSION: HPM treatment can regulate intestinal flora and inhibit the expression of TNF-alpha and IL-12 in the colon tissues of UC rats, indicating that HPM can improve colonic immune response. (C) 2012 Baishideng. All rights reserved.