Ramamoorthy Prabhu

Oral glutamine attenuates surgical manipulation-induced alterations in the intestinal brush border membrane.

BACKGROUND Our earlier work has shown that surgical manipulation of the intestine results in oxidative stress and mucosal damage along with alterations in the brush border membrane (BBM). Glutamine feeding is known to offer protection against damage to mucosa under various conditions and this study looked at the effect of oral supplementation of glutamine or glutamic acid in the intestinal BBM alterations after surgical manipulation. MATERIALS AND METHODS Control and rats pretreated for 7 days with 2% glutamine or glutamic acid or isonitrogenous amino acids, glycine, or alanine were subjected to surgical manipulation of the intestine. BBMs were isolated from the intestine and functional and structural alterations to these membranes were assessed and compared. RESULTS Surgical manipulation resulted in oxidative stress in the enterocyte BBM and these changes included a decrease in alkaline phosphatase activity and alpha-tocopherol content along with an increase in lipid peroxidation parameters. A decrease in glucose transport by the isolated BBM vesicles suggested functional impairment. Surgical manipulation also resulted in phospholipid degradation possibly mediated by PLA(2) and membrane protease activation. Glutamine or glutamic acid supplementation prevented these changes but not by glycine or alanine. CONCLUSION This study suggests that oral glutamine or glutamic acid supplementation prior to surgery can offer protection to the intestine and this might prevent postsurgical complications.

Retinoid metabolism in the rat small intestine.

Vitamin A (retinol) is essential for epithelial cell growth, differentiation and proliferation. The absorption of retinol occurs in the small intestine, and the metabolism of this vitamin is not well studied in this organ. The intestinal epithelium has a high rate of cell proliferation and differentiation, and the present study looked at the level of retinoids and metabolizing enzymes involved in their interconversion along the villus-crypt axis under normal conditions. Intestine was removed from control rats, and enterocytes at various stages of maturation and differentiation were quantified by the metal chelation method. Using HPLC, various retinoid concentrations in the cell homogenate and the metabolizing enzymes in the cytosol were quantified. The proliferating crypt cells were found to have a higher level of retinoic acid as well as of the enzymes involved in its formation, such as retinaldehyde oxidase and retinol dehydrogenase, compared with the villus cells, suggesting a possible role for this compound in intestinal epithelial cell proliferation and differentiation. The high level of retinol and high retinaldehyde reductase activity in the villus cells suggest the important role played by this enzyme in the conversion of dietary beta-carotene to retinol via retinaldehyde. In summary, this study has given for the first time a detailed analysis of the retinoid levels and metabolizing enzymes in different cell populations in the rat small intestinal epithelium.

Simple method of preparation of rat colonocyte apical membranes using polyethylene glycol precipitation

Background: Although a number of methods are available for the preparation of brush border membranes (BBM) from the small intestine, very few studies have been performed on the isolation of colonic apical membranes (CAM). Cation precipitation is one of the methods used for the isolation of these membranes, which can result in altered lipid composition as a result of activation of phospholipase. We have earlier established a method for BBM isolation using polyethylene glycol (PEG) precipitation. In the present study, CAM from isolated colonocytes were prepared using PEG and the isolated membranes were characterized.