Citation

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THE LIPOPOLYSACCHARIDE DETOXIFYING PROPERTIES OF CALF INTESTINAL ALKALINE PHOSPHATASE: AN IN VITRO STUDY

Bouwman, C;

This thesis provides an overview of the role of intestinal alkaline phosphatase (IAP) and lipopolysaccharide (LPS) in liver fibrosis and investigates the anti-inflammatory properties of IAP in vitro. IAP is an enzyme primarily expressed in the intestines, playing a vital role in nutrient absorption and gut health. LPS, an endotoxin found in gram-negative bacteria, triggers an inflammatory response when leaked from the intestines into the portal vein and subsequently into the liver. In liver fibrosis, LPS-induced inflammation leads to excessive extracellular matrix production and fibrotic tissue formation. Studies suggest that IAP can detoxify LPS by dephosphorylating its lipid A component. The thesis focuses on exploring additional factors that may enhance the detoxification of LPS by IAP through nitric oxide (NO) assays, p-nitrophenyl phosphate (pNPP) assays and co-immunoprecipitation. IAP was unable to significantly reduce inflammation caused by LPS on RAW 264.7 cells. With pNPP assays it was found that IAP had a higher activity in deproteinized and regular fetal bovine serum (FBS), compared to delipidated FBS. Additionally, during the immunoprecipitation experiment, several proteins co-eluted with IAP and IAP+LPS from FBS. There was no big visual difference on the SDS-page gel between the sample containing IAP compared to IAP + LPS. Further analysis using LC-MS is required to identify and quantify these co-eluted proteins of interest. These findings suggest that lipids and proteins in the cellular environment may play a role in modulating the activity of AP.