Functional Activity of Renal Cytochrome P-450 1A1 Enzyme in Rats with Low Birth-Weight
Date of Award
Restricted Access Thesis
College of Theology, Arts, & Sciences
Math & Science
Dr. Rici Hallstrand
To determine the effect of maternal low protein diet (LPD) administered during pregnancy on the status of renal cytochrome-P450 (CYP) enzymes in the offspring, pregnant rats were fed either a purified control diet (C76) or a low protein diet (L76) throughout pregnancy and lactation and offspring were weaned onto lab chow on postnatal day 28. The control diet contained 18% protein while the LPD contained 8% protein. One male and one female offspring were sacrificed and kidneys were collected and frozen from 28-day, 65-day, and 150-day old rats. Renal microsomes were prepared via differential centrifugation technique. For the current study, we examined CYP1A1 which is an important drug metabolizing enzyme involved in the generation of carcinogenic metabolites. Its main role is the oxidation of aromatic amines, polarizing such compounds so they may be readily metabolized and excreted from the body. CYP1A2 activity was measured via its ability to oxidize the substrate ethoxy-resorufin into the fluorescent product resorufin; activity was measured at 530/590 nm upon incubation of renal microsomes for 30 min. Preliminary analysis of day 28, 65, and 150 male and female rats suggests no alterations in CYP1A2 activity normalized to microsomal protein amount. However, upon normalization to total kidney mass, we found significant changes. In summary, renal CYP1A1 activity is permanently altered by early life nutritional insult.
Honors: Thesis with Distinction Award