Objectives: Congenital hepatic fibrosis (CHF) is an important cause of morbidity and mortality in patients with autosomal recessive polycystic kidney disease (ARPKD). The pathogenesis of CHF remains undefined. Several recent studies suggest that the renin-angiotensin system (RAS) is an important mediator of progressive hepatic fibrosis through activation of profibrotic mediators, such as transforming growth factor-β (TGF-β). RAS activation has not previously been studied in patients with CHF or in animal models. The aim of the present study was to characterize RAS expression during the course of CHF in the PCK rat.
Materials and Methods: Studies were conducted in the PCK rat, an orthologous ARPKD/CHF model, and age-matched normal control Sprague-Dawley rats. Expression of the RAS components, renin, angiotensinogen, angiotensin-converting enzyme (ACE), and angiotensin II type 1 receptor (AT1R), as well as the profibrotic mediator TGF-β, was examined in cystic PCK and control rat livers at 2, 4, and 6 months of age by quantitative real-time polymerase chain rection (qRT-PCR). Angiotensin II (ANG II) was examined by immunohistochemistry (IHC). Fibrosis was assessed by IHC using reticulin staining and Masson trichrome. Collagen content was determined by hydroxyproline analysis.
Results: Progressive fibrosis and increased hepatic collagen content occurred in PCK rats with age. In 4- and 6-month-old PCK rat livers, ACE gene expression was markedly increased, 8- and 17-fold, respectively, compared with age-matched control livers. Expression of the other RAS components, renin, angiotensinogen, and AT1R were not significantly different. IHC demonstrated prominent ANG II protein expression in periportal regions in PCK rats. In contrast, no expression was noted in control livers. TGF-β expression was also increased in PCK rat livers with progressive disease.
Conclusions: The present study demonstrates, for the first time, RAS upregulation in an orthologous rat ARPKD/CHF model. Increases in ACE and ANG II, as well as the downstream target, the profibrotic mediator TGF-β, suggest that RAS activation may be an important mediator of CHF disease progression. The findings also suggest that treatment with RAS inhibitors, specifically ACE inhibitors or AT1R blockers, could be therapeutic in slowing disease progression in CHF.
*Department of Research, MetroHealth Medical Center, Case Western Reserve University, Cleveland, OH, USA
‡Division of Gastroenterology, Hepatology, and Nutrition, the Children's Hospital of Philadelphia, Philadelphia, USA
§Departments of Medicine and Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, USA.
Received 4 June, 2009
Accepted 10 November, 2009
Address correspondence and reprint requests to Katherine MacRae Dell, MD, Department of Research, MetroHealth Medical Center, 2500 MetroHealth Dr, R457, Cleveland, OH 44109 (e-mail: KatherineDell@case.edu).
This research was supported by the NIH/NIDDK (DK-058123 to R.G.W.), the National Kidney Foundation of Ohio, MetroHealth Medical Center, and a pilot award from the UAB Recessive PKD Center Core (to K.M.D.). R.G.W. is funded in part by a pilot award from the ARPKD/CHF Alliance. J.W. is supported by NIH training grant T32-DK007006.
The authors report no conflicts of interest.