Proceedings of The Physiological Society
University of Manchester (2010) Proc Physiol Soc 19, PC294
The effect of acetylsalicylic acid on early renal development using cultured embryonic metanephros.
A. S. Carvalho1, M. J. Elmes1, S. J. Welham1
1. Nutritional Sciences, University of Nottingham, Loughborough, United Kingdom.
The maternal ingestion of Non-Steroidal Anti-Inflammatory drugs (NSAIDs) during pregnancy has been associated with renal failure and abnormal renal development in the offspring at birth and throughout life. The objective of this study was to evaluate the effects of the non-selective NSAID acetylsalicylic acid (ASA), also known as Aspirin, on ex vivo embryonic kidney growth and development. Dams were mated and 12 days after plugging, at day 12 of gestation, were culled. Embryos were quickly removed and decapitated. Whole metanephroi (between n=8 and n=12) were microdissected from the embryos and culture for 7 days in DMEM: F-12 medium. The contra-laterals were cultured in concentrations of ASA approximate to physiological doses; ranging from 0.04mg/mL-0.4mg/mL, where 0.2mg/mL is equivalent to taking two 500mg tablets of Aspirin. Images were taken at 6x magnification on the day of dissection (day 0) and every 24hours throughout the experiment. Metanephroi cross-sectional areas were measured using Image Pro Plus v5.1. Using a paired t-test, the data showed that ASA concentrations as low as 0.1mg/mL produced significant smaller metanephroi compared with the control, while the concentration of 0.4mg/mL had a detrimental effect on size as well as structure. Metanephroi cultured with 0.1mg/ml of ASA incurred a significant decrease in size by day 4 of culture (P<0.025), the physiological dose of 0.2mg/mL was significantly smaller by days 3 (P<0.05), and 0.4mg/mL resulted in significantly smaller metanephroi from day 1 (P<0.005). The results suggest that ASA has an adverse effect on early stage renal growth and development. We hypothesise that ASA is causing this effect through its inhibitory properties on the COX enzymes, which are responsible for the conversion of arachidonic acid to prostaglandins, hence implicating a role for prostaglandins in renal development. Future work for this study includes attempting to rescue the cultured metanephroi with exogenous prostaglandins to determine whether this is the mechanism involved in producing the phenotypes observed, and also to analyse the pH of ASA at the different concentration to rule out acidity as a factor.
Where applicable, experiments conform with Society ethical requirements