K+‐channel inhibition reduces portal perfusion pressure in fibrotic rats and fibrosis associated characteristics of hepatic stellate cells

Liver International - Tập 35 Số 4 - Trang 1244-1252 - 2015
Christian Freise1, Silke Heldwein2,1, Ulrike Erben1, Joachim Hoyer3, Ralf Köhler4, Korinna Jöhrens5, Eleonora Patsenker6, Martin Rühl1, Daniel Seehofer7, Felix Stickel6, Rajan Somasundaram1
1Department of Gastroenterology Infectiology and Rheumatology Charité – University Medicine Berlin Campus Benjamin Franklin Berlin Germany
2Department of Clinical Research ‐ Hepatology University of Berne Berne Switzerland
3Department of Internal Medicine and Nephrology, University Hospital, Philipps-University, Marburg, Germany
4I+CS and ARAID Aragon Institute of Health Sciences Zaragoza 50009 Spain
5Department of Pathology Charité ‐ University Medicine Berlin Campus Charité Mitte Berlin Germany
6Department of Clinical Research - Hepatology; University of Berne; Berne Switzerland
7Department of General Visceral and Transplantation Surgery Charité – University Medicine Berlin Campus Virchow Klinikum Berlin Germany

Tóm tắt

AbstractBackground & AimsIn liver fibrosis, activated hepatic stellate cells (HSC) secrete excess extracellular matrix, thus, represent key targets for antifibrotic treatment strategies. Intermediate‐conductance Ca2 + ‐activated K+‐channels (KCa3.1) are expressed in non‐excitable tissues affecting proliferation, migration and vascular resistance rendering KCa3.1 potential targets in liver fibrosis. So far, no information about KCa3.1 expression and their role in HSC exists. Aim was to quantify the KCa3.1 expression in HSC depending on HSC activation and investigation of antifibrotic properties of the specific KCa3.1 inhibitor TRAM‐34 in vitro and in vivo.MethodsKCa3.1 expression and functionality were studied in TGF‐β1–activated HSC by quantitative real time PCR, western‐blot and patch‐clamp analysis respectively. Effects of TRAM‐34 on HSC proliferation, cell cycle and fibrosis‐related gene expression were assessed by [3H]‐thymidine incorporation, FACS‐analysis and RT‐PCR respectively. In vivo, vascular resistance and KCa3.1 gene and protein expression were determined in bile duct ligated rats by in situ liver perfusion, Taqman PCR and immunohistochemistry respectively.ResultsFibrotic tissues and TGF‐β1‐activated HSC exhibited higher KCa3.1‐expressions than normal tissue and untreated cells. KCa3.1 inhibition with TRAM‐34 reduced HSC proliferation by induction of cell cycle arrest and reduced TGF‐β1‐induced gene expression of collagen I, alpha‐smooth muscle actin and TGF‐β1 itself. Furthermore, TRAM‐34 blocked TGF‐β1‐induced activation of TGF‐β signalling in HSC. In vivo, TRAM‐34 reduced the thromboxane agonist‐induced portal perfusion pressure.ConclusionInhibition of KCa3.1 with TRAM‐34 downregulates fibrosis‐associated gene expression in vitro, and reduces portal perfusion pressure in vivo. Thus, KCa3.1 may represent novel targets for the treatment of liver fibrosis.

Từ khóa


Tài liệu tham khảo

10.1172/JCI24282

10.1172/JCI66028

10.1016/S0168-8278(02)00263-5

10.1016/j.dld.2004.01.003

10.1055/s-2001-17550

10.1016/j.jhep.2006.04.014

Tuchweber B, 1996, Proliferation and phenotypic modulation of portal fibroblasts in the early stages of cholestatic fibrosis in the rat, Lab Invest, 74, 265

10.1074/jbc.M700194200

10.1053/jhep.2001.28457

10.1038/ncpgasthep0055

10.1055/s-2001-17558

10.1016/S0399-8320(07)73970-2

10.2741/reeves

Beljaars L, 2002, Targeting hepatic stellate cells for cell‐specific treatment of liver fibrosis, Front Biosci, 7, e214, 10.2741/beljaars

10.1159/000090320

10.1590/S0102-86502006000700013

10.1016/S0168-8278(98)80057-3

10.1053/jhep.2001.23500

10.1016/S0168-8278(01)00198-2

10.3349/ymj.2004.45.4.649

Marra F, 2002, Role of hepatic stellate cells in the pathogenesis of portal hypertension, Nefrologia, 22, 34

10.2174/1389450013348173

10.1161/01.RES.85.9.e33

10.1007/s002329900601

Wulff H, 2003, Potassium channels as therapeutic targets for autoimmune disorders, Curr Opin Drug Discov Devel, 6, 640

10.4049/jimmunol.173.2.776

Wulff H, 2007, Targeting effector memory T‐cells with Kv1.3 blockers, Curr Opin Drug Discov Devel, 10, 438

10.1161/01.ATV.0000156399.12787.5c

10.1073/pnas.0903458106

10.1073/pnas.97.14.8151

Rojkind M, 1995, Characterization and functional studies on rat liver fat‐storing cell line and freshly isolated hepatocyte coculture system, Am J Pathol, 146, 1508

Weiskirchen R, 2005, Isolation and culture of hepatic stellate cells, Methods Mol Med, 117, 99

10.1016/j.phrs.2014.04.013

10.1016/j.jhep.2007.03.019

10.1016/j.jhep.2010.10.044

10.1016/B978-0-12-091302-2.50010-4

10.1016/S0168-8278(00)80422-5

10.1152/ajpgi.00315.2002

10.1007/s002329900135

10.1016/j.bbrc.2004.02.041

10.1002/cphy.c120035

10.2337/db13-0135

10.1093/ndt/gft431

10.1016/j.tox.2008.06.013

10.1111/j.1478-3231.2008.01668.x

10.1152/ajpgi.00337.2005

10.1172/JCI115905

Rockey DC, 1992, Rat hepatic lipocytes express smooth muscle actin upon activation in vivo and in culture, J Submicrosc Cytol Pathol, 24, 193

10.1136/gut.50.4.571

10.1016/S1089-3261(05)70195-1

10.1053/jhep.2003.50044

10.1371/journal.pone.0085244

10.1371/journal.pone.0063028

Thông tin
Thông tin xuất bản

Liver International

Tập 35 Số 4

1244-1252

Thông tin tác giả