PMID: 9537853Apr 16, 1998Paper

Detection of endothelin-1 and its receptors in rat liver endothelial cells by in situ reverse transcriptase-polymerase chain reaction

Journal of Hepatology
I DiamantisJ Reichen

Abstract

Endothelin (ET)-1 is a potent vasoconstrictor, also in the liver; it increases intrahepatic resistance by targeting hepatic stellate cells acting via the ET-A receptor. The role of the expression and regulation of ET-1 and its receptors in endothelial liver cells, an important site of ET production in normal liver, is not yet well documented. In the present study, we have developed an in situ reverse transcriptase (RT)-polymerase chain reaction method to elucidate the presence of prepro-ET-1, ET-A and ET-B receptors in isolated rat liver endothelial cells. After in situ collagenase-pronase liver perfusion, endothelial cells were isolated using a Nycodenz gradient. RT-polymerase chain reaction and in situ RT-polymerase chain reaction were performed using specific primers for prepro-endothelin, ET-A and ET-B. Trypan blue exclusion test showed a viability of more than 90% in the freshly isolated non-parenchymal cells. RT-polymerase chain reaction showed expression of prepro-endothelin, ET-A and ET-B in RNA isolated from the endothelial cells. After in situ RT-polymerase chain reaction, we detected cytoplasmatic staining representing the presence of mRNA in the endothelial cells, with all sets of primers. About 60-80% of cells were...Continue Reading

References

Aug 10, 1995·The New England Journal of Medicine·E R Levin
Jan 1, 1994·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·S OkumuraT Kashiwagi
Oct 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·C HoussetD M Bissell
Mar 8, 1993·FEBS Letters·T A Tran-ThiK Decker
May 1, 1997·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·M B CastilloM R Celio

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Citations

May 20, 2011·Journal of Gastroenterology and Hepatology·Wei ChenTingbo Liang
Dec 20, 2008·Acta Anaesthesiologica Scandinavica·A AnemanA Hadengue
May 16, 2002·Journal of Cellular Physiology·Rong Shao, Don C Rockey

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