Activation of Trpv4 Reduces the Hyperproliferative Phenotype of Cystic Cholangiocytes From an Animal Model of ARPKD

被引：82

作者：

Gradilone, Sergio A. ^{[1
]}

Masyuk, Tatyana V. ^{[1
]}

Huang, Bing Q. ^{[1
]}

Banales, Jesus M. ^{[1
,2
,3
]}

Lehmann, Guillermo L. ^{[1
,4
]}

Radtke, Brynn N. ^{[1
]}

Stroope, Angela ^{[1
]}

Masyuk, Anatoliy I. ^{[1
]}

Splinter, Patrick L. ^{[1
]}

LaRusso, Nicholas F. ^{[1
]}

机构：

[1] Mayo Clin, Coll Med, Div Gastroenterol & Hepatol, Miles & Shirley Fiterman Ctr Digest Dis, Rochester, MN 55905 USA

[2] Univ Navarra & Ciberehd, Mol Genet Lab, Div Gene Therapy & Hepatol, Sch Med, Pamplona, Spain

[3] Univ Navarra & Ciberehd, CIMA, Pamplona, Spain

[4] Univ Nacl Rosario, Fac Ciencias Bioquim & Farmaceut, Consejo Nacl Invest Cient & Tecn, Inst Fisiol Expt, RA-2000 Rosario, Santa Fe, Argentina

来源：

GASTROENTEROLOGY | 2010年 / 139卷 / 01期

基金：

美国国家卫生研究院;

关键词：

Calcium; PKD; Cholangiocytes; Trpv4; POLYCYSTIC KIDNEY-DISEASE; RECEPTOR POTENTIAL VANILLOID-4; CATION CHANNEL TRPV4; AUTOSOMAL-DOMINANT; LIVER-DISEASE; EPITHELIAL-CELLS; HEPATIC CYSTOGENESIS; PCK RAT; B-RAF; PROTEIN;

D O I：

10.1053/j.gastro.2010.04.010

中图分类号：

R57 [消化系及腹部疾病];

学科分类号：

摘要：

BACKGROUND & AIMS: In polycystic liver diseases, cyst formation involves cholangiocyte hyperproliferation. In polycystic kidney (PCK) rats, an animal model of autosomal-recessive polycystic kidney disease (ARPKD), decreased intracellular calcium [Ca2+](i) in cholangiocytes is associated with hyperproliferation. We recently showed transient receptor potential vanilloid 4 (Trpv4), a calcium-entry channel, is expressed in normal cholangiocytes and its activation leads to [Ca2+](i) increase. Thus, we hypothesized that pharmacologic activation of Trpv4 might reverse the hyperproliferative phenotype of PCK cholangiocytes. METHODS: Trpv4 expression was examined in liver of normal and PCK rats, normal human beings, and patients with autosomal-dominant polycystic kidney disease or ARPKD. Trpv4 activation effect on cell proliferation and cyst formation was assessed in cholangiocytes derived from normal and PCK rats. The in vivo effects of Trpv4 activation on kidney and liver cysts was analyzed in PCK rats. RESULTS: Trpv4 was overexpressed both at messenger RNA (8-fold) and protein (3-fold) levels in PCK cholangiocytes. Confocal and immunogold electron microscopy supported Trpv4 overexpression in the livers of PCK rats and ARPKD or autosomal-dominant polycystic kidney disease patients. Trpv4 activation in PCK cholangiocytes increased [Ca2+](i) by 30%, inhibiting cell proliferation by approximately 25%-50% and cyst growth in 3-dimensional culture (3-fold). Trpv4-small interfering RNA silencing blocked effects of Trpv4 activators by 70%. Trpv4 activation was associated with Akt phosphorylation and beta-Raf and Erk1/2 inhibition. In vivo, Trpv4 activation induced a significant decrease in renal cystic area and a nonsignificant decrease in liver cysts. CONCLUSIONS: Taken together, our in vitro and in vivo data suggest that increasing intracellular calcium by Trpv4 activation may represent a potential therapeutic approach in PKD.

引用

页码：304 / U425

页数：13

共 56 条

[1] Clinical and molecular characterization defines a broadened spectrum of autosomal recessive polycystic kidney disease (ARPKD) [J].