OBJECTIVE: Irritable bowel syndrome (IBS) is a common disorder in clinical practice, but the pathophysiology of IBS has not been completely elucidated yet. Experiments have revealed that the concentrations of some kinds of brain-gut peptides, such as substance P, were abnormal in the plasma and/or the intestinal mucosa. In order to explore further the possible role of substance P containing nerve fibers in the enteric nervous system and central nervous system, the expression of c-fos, a well-established marker of activated neural pathway, was induced to show substance P containing a neural pathway in the rat model of constipation-predominant IBS by rectal distention. METHODS: The rat model was set up by intragastric instillation of 2.0 mL water at 0-4 degrees C in 20 male Wistar rats for two weeks. Both the model group and the controls underwent rectal distention under deep anesthesia. Sections containing the anatomical areas of interest were obtained and processed for c-fos protein and substance P immunohistochemistry using the strept avidin-biotin complex (SABC) method. The staining results were analyzed semi-quantitatively, using a computerized color image analyzer with two parameters: opacity density and immunoreactive areas. The statistical difference of the opacity density and immunoreactive areas between the two groups was analyzed by a t-test. Correlation analysis was used to investigate the relationship between the expression of substance P and c-fos protein of the same region in the model group. RESULTS: The opacity density of substance P immunoreactive tissues in the ileocecal junction, colon, the posterior horn of the spinal cord and the hypothalamus of the model group were all significantly higher compared with those in the control group (176.6 vs 155.5, 172.3 vs 152.0, 182.1 vs 160.2, 128.3 vs 117.9; P < 0.05, respectively). Meanwhile in the ileocecal junction, colon, the posterior horn of the spinal cord and the hypothalamus of the model group, the opacity density of c-fos protein-positive tissue were all significantly higher than those of the same region in the controls (120.9 vs 109.0, 101.3 vs 92.2, 125.4 vs 88.7, 115.5 vs 88.6; P < 0.05, respectively). The distribution of c-fos protein-positive tissue is similar to that of the substance P and the analysis shows that there is close correlation between the expression of substance P and c-fos protein of the same region in the model group (r = 0.594-0.721, P < 0.05). CONCLUSIONS: The expression of substance P and c-fos protein in both the enteric nervous system and the central nervous system of the constipation-predominant IBS rat model is abnormal, which suggests that an abnormal change in substance P may be involved in the pathogenesis of IBS and the substance P-containing neural pathway may be one of the neural pathways that play important role in the regulation of the gastrointestinal function.
OBJECTIVE:Irritable bowel syndrome (IBS) is a common disorder in clinical practice, but the pathophysiology of IBS has not been completely elucidated yet. Experiments have revealed that the concentrations of some kinds of brain-gut peptides, such as substance P, were abnormal in the plasma and/or the intestinal mucosa. In order to explore further the possible role of substance P containing nerve fibers in the enteric nervous system and central nervous system, the expression of c-fos, a well-established marker of activated neural pathway, was induced to show substance P containing a neural pathway in the rat model of constipation-predominant IBS by rectal distention. METHODS: The rat model was set up by intragastric instillation of 2.0 mL water at 0-4 degrees C in 20 male Wistar rats for two weeks. Both the model group and the controls underwent rectal distention under deep anesthesia. Sections containing the anatomical areas of interest were obtained and processed for c-fos protein and substance P immunohistochemistry using the strept avidin-biotin complex (SABC) method. The staining results were analyzed semi-quantitatively, using a computerized color image analyzer with two parameters: opacity density and immunoreactive areas. The statistical difference of the opacity density and immunoreactive areas between the two groups was analyzed by a t-test. Correlation analysis was used to investigate the relationship between the expression of substance P and c-fos protein of the same region in the model group. RESULTS: The opacity density of substance P immunoreactive tissues in the ileocecal junction, colon, the posterior horn of the spinal cord and the hypothalamus of the model group were all significantly higher compared with those in the control group (176.6 vs 155.5, 172.3 vs 152.0, 182.1 vs 160.2, 128.3 vs 117.9; P < 0.05, respectively). Meanwhile in the ileocecal junction, colon, the posterior horn of the spinal cord and the hypothalamus of the model group, the opacity density of c-fos protein-positive tissue were all significantly higher than those of the same region in the controls (120.9 vs 109.0, 101.3 vs 92.2, 125.4 vs 88.7, 115.5 vs 88.6; P < 0.05, respectively). The distribution of c-fos protein-positive tissue is similar to that of the substance P and the analysis shows that there is close correlation between the expression of substance P and c-fos protein of the same region in the model group (r = 0.594-0.721, P < 0.05). CONCLUSIONS: The expression of substance P and c-fos protein in both the enteric nervous system and the central nervous system of the constipation-predominant IBSrat model is abnormal, which suggests that an abnormal change in substance P may be involved in the pathogenesis of IBS and the substance P-containing neural pathway may be one of the neural pathways that play important role in the regulation of the gastrointestinal function.