Glibenclamide, ATP and Metformin Increases the Expression of Human Bile Salt Export Pump ABCB11

ABSTRACT Background Bile Salt Export Pump (BSEP/ABCB11) is important in the maintenance of the enterohepatic circulation of bile acids and drugs. Drugs such as rifampicin, glibenclamide inhibit BSEP. Progressive Familial Intrahepatic Cholestasis Type-2, a lethal pediatric disease, some forms of intrahepatic cholestasis of pregnancy, and drug-induced cholestasis are associated with BSEP dysfunction. Methods We started with a bioinformatic approach to identify the relationship between ABCB11 and other proteins, microRNAs, and drugs. Microarray data set of the liver samples from ABCB11 knockout mice was analyzed by GEO2R tool. Differentially expressed gene pathway enrichment analysis was done by ClueGo v2.5.5 app from Cytoscape. Protein-protein interaction network was constructed by STRING application in Cytoscape. Networks were analyzed using the Cytoscape software v3.7.1. CyTargetLinker v4.1.0 was used to screen the transcription factors, microRNAs and drugs. Predicted drugs were validated on human liver cell line, HepG2. BSEP expression was quantified by Real Time PCR and Western Blot. Results ABCB11 knockout in mice was associated with a predominant upregulation and downregulation of genes associated with cellular component movement and sterol metabolism respectively. We further identified the hub genes in the network. Genes related to immune activity, cell signaling and fatty acid metabolism were dysregulated. We further identified drugs (glibenclamide and ATP) and a total of 14 microRNAs targeting the gene. Western Blot and Real Time PCR analysis confirmed the upregulation of BSEP on the treatment of HepG2 cells with glibenclamide, ATP, and metformin. Conclusion The differential expression of cell signaling genes and those related to immune activity in ABCB11 KO animals may be secondary to cell injury. We have found glibenclamide, ATP, and metformin upregulates BSEP. The mechanisms involved and the clinical relevance of these findings need to be investigated.