J Jørgensen1, P B Mortensen. 1. Dept. of Medicine, Copenhagen University Hospital, Rigshospitalet, Denmark.
Abstract
BACKGROUND: Previous metabolic studies of the colonic mucosa have been done using isolated cells or small biopsy specimens. METHODS: A new method for assessing the utilization of short-chain fatty acids in human colonic mucosal tissue strips considerably larger than routine samples was evaluated and compared with the method of isolated colonocytes. Human colonic mucosal strips and isolated human and rat colonocytes were incubated with acetate (C2), butyrate (C4), and hexanoate (C6), and oxidation rates obtained by quantifying the production of CO2. RESULTS: The wet weight of strips was highly correlated with the production of CO2, and intersample coefficient of variance was <10%. The production of CO2 from the oxidation of C2, C4, and C6 was in the order of C2 > C4 > C6 for both strips and isolated human and rat colonocytes. The production of adenosine triphosphate (ATP) in strips and isolated human and rat colonocytes was in the order of C2 < or = C4 < or = C6. The Km value for the oxidation of butyrate to CO2 in strips (1.8 mmol/l) was several times higher than previously reported for isolated human and rat colonocytes (0.1-0.3 mmol/l). CONCLUSIONS: This new method is highly reproducible and able to assess the metabolic activity of the colonic mucosa. The high Km value of butyrate oxidation in mucosal strips seems to reflect the in vivo Km value of colonocytes and shows the importance of a preserved anatomic structure in metabolic studies of the colonic epithelium. The low Km value for isolated colonocytes probably reflects the intracellular ability to oxidize butyrate. We propose that both isolated colonocytes and mucosal strips be used in studies of colonic mucosal metabolism. This method is relevant in disease states of the colon in which a disagreement prevails as to the ability to oxidize butyrate by colonocytes, such as in ulcerative colitis.
BACKGROUND: Previous metabolic studies of the colonic mucosa have been done using isolated cells or small biopsy specimens. METHODS: A new method for assessing the utilization of short-chain fatty acids in humancolonic mucosal tissue strips considerably larger than routine samples was evaluated and compared with the method of isolated colonocytes. Humancolonic mucosal strips and isolated human and rat colonocytes were incubated with acetate (C2), butyrate (C4), and hexanoate (C6), and oxidation rates obtained by quantifying the production of CO2. RESULTS: The wet weight of strips was highly correlated with the production of CO2, and intersample coefficient of variance was <10%. The production of CO2 from the oxidation of C2, C4, and C6 was in the order of C2 > C4 > C6 for both strips and isolated human and rat colonocytes. The production of adenosine triphosphate (ATP) in strips and isolated human and rat colonocytes was in the order of C2 < or = C4 < or = C6. The Km value for the oxidation of butyrate to CO2 in strips (1.8 mmol/l) was several times higher than previously reported for isolated human and rat colonocytes (0.1-0.3 mmol/l). CONCLUSIONS: This new method is highly reproducible and able to assess the metabolic activity of the colonic mucosa. The high Km value of butyrate oxidation in mucosal strips seems to reflect the in vivo Km value of colonocytes and shows the importance of a preserved anatomic structure in metabolic studies of the colonic epithelium. The low Km value for isolated colonocytes probably reflects the intracellular ability to oxidize butyrate. We propose that both isolated colonocytes and mucosal strips be used in studies of colonic mucosal metabolism. This method is relevant in disease states of the colon in which a disagreement prevails as to the ability to oxidize butyrate by colonocytes, such as in ulcerative colitis.