Objective Aldosterone, one of the main peptides in renin angiotensin aldosterone system (RAAS), has been suggested to mediate liver fibrosis and portal hypertension. Spironolactone, an aldosterone antagonist, has beneficial effect on hyperdynamic circulation in clinical practice. However, the mechanisms remain unclear. The present study aimed to investigate the role of spionolactone on liver cirrhosis and portal hypertension. Methods Liver cirrhosis was induced by bile duct ligation (BDL). Spironolactone was administered orally (20 mg/kg/d) after bile duct ligation was performed. Liver fibrosis was assessed by histology, Masson's trichrome staining, and the measurement of hydroxyproline and type I collagen content. The activation of HSC was determined by analysis of alpha smooth muscle actin (α-SMA) expression. Protein expressions and protein phosphorylation were determined by immunohistochemical staining and Western blot analysis, Messenger RNA levels by quantitative real time polymerase chain reaction (Q-PCR). Portal pressure and intrahepatic resistance were examined in vivo. Results Treatment with spironolactone significantly lowered portal pressure. This was associated with attenuation of liver fibrosis, intrahepatic resistance and inhibition of HSC activation. In BDL rat liver, spironolactone suppressed up-regulation of proinflammatory cytokines (TNFα and IL-6). Additionally, spironolactone significantly decreased ROCK-2 activity without affecting expression of RhoA and Ras. Moreover, spironolactone markedly increased the levels of endothelial nitric oxide synthase (eNOS), phosphorylated eNOS and the activity of NO effector- protein kinase G (PKG) in the liver. Conclusion Spironolactone lowers portal hypertension by improvement of liver fibrosis and inhibition of intrahepatic vasoconstriction via down-regulating ROCK-2 activity and activating NO/PKG pathway. Thus, early spironolactone therapy might be the optional therapy in cirrhosis and portal hypertension.
References
[1]
Bosch J, Garcia-Pagan JC (2000) Complications of cirrhosis. I. Portal hypertension. J Hepatol 32: 141–156.
[2]
Groszmann RJ, Abraldes JG (2005) Portal hypertension: from bedside to bench. J Clin Gastroenterol 39: S125–S130.
[3]
Rodriguez-Vilarrupla A, Fernandez M, Bosch J, Garcia- Pagan JC (2007) Current concepts on the pathophysiology of portal hypertension. Ann Hepatol 6: 28–36.
[4]
Friedman SL (2008) Mechanisms of hepatic fibrogenesis. Gastroenterology 134: 1655–1669.
[5]
Trebicka J, Hennenberg M, Laleman W, Shelest N, Biecker E, et al. (2007) Atorvastatin lowers in portal pressure in cirrhotic rats by inhibition of RhoA/Rho-kinase and activation of endothelial nitric oxide synthase. Hepatology 46: 242–253.
[6]
Sowers JR, Whaley-Connell A, Epstein M (2009) Narrative review: the emerging clinical implications of the role of aldosterone in the metabolic syndrome and resistant hypertension. Ann Intern Med 150: 776–783.
[7]
Li X, Meng Y, Wu P, Zhang Z, Yang X (2009) Angiotensin II and aldosterone stimulating NF-κB and AP-1 activation in hepatic fibrosis of rat. Regul Pept 138: 15–25.
[8]
Fujisawa G, Muto S, Okada K, Kusano E, Ishibashi S (2006) Mineralocorticoid receptor antagonist spironolactone prevents pig serum-induced hepatic fibrosis in rats. Transl Res 148: 149–156.
[9]
Tandon P, Abraldes JG, Berzigotti A, Garcia-Pagan JC, Bosch J (2010) Renin-angiotensin-aldosterone inhibitors in the reduction of portal pressure: a systematic review and meta-analysis. J Hepatol 53: 273–282.
[10]
Wiest R, Groszmann RJ (2002) The paradox of nitric oxide in cirrhosis and portal hypertension: too much, not enough. Hepatology 35: 478–491.
[11]
Ji H, Meng Y, Zhang X, Luo W, Wu P, et al. (2011) Aldosterone induction of hepatic stellate cell contraction through activation of RhoA/ROCK-2 signaling pathway. Regul Pept 169: 13–20.
[12]
Paizis G, Cooper ME, Schembri JM, Tikellis C, Burrell LM, et al. (2002) Up-regulation of components of the renin-angiotensin system in the bile duct-ligated rat liver. Gastroenterology 123: 1667–1676.
[13]
Kountouras J, Billing BH, Scheuer PJ (1984) Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat. Br J Exp Pathol 65: 305–311.
[14]
Li X, Yang X, Wu P, Meng Y, Li S, et al. (2001) Gene-CYP11B2 expression in rat liver in hepatic fibrogenesis induced by CCl4. Chin Med J (Engl) 114(1): 64–68.
[15]
Trebicka J, Leifeld L, Hennenberg M, Biecker E, Eckhardt A, et al. (2008) Hemodynamic effects of urotensin II and its specific receptor antagonist palosuran in cirrhotic rats. Hepatology 47: 1264–1276.
[16]
Zhang XL, Liu JM, Yang CC, Zheng YL, Liu L, et al. (2006) Dynamic expression of extracellular signal-regulated kinase in rat liver tissue during hepatic fibrogenesis. World J Gastroenterol 12: 6376–6381.
[17]
Miyoshi H, Rust C, Roberts PJ, Burgart LJ, Gores GJ (1999) Hepatocyte apoptosis after bile duct ligation in the mouse involves Fas. Gastroenterology 117: 669–677.
[18]
Oelze M, Mollnau H, Hoffmann N, Warnholtz A, Bodenschatz M, et al. (2000) Vasodilator-stimulated phosphoprotein serine 239 phosphorylation as a sensitive monitor of defective nitric oxide/cGMP signaling and endothelial dysfunction. Circ Res 87: 999–1005.
[19]
Hennenberg M, Biecker E, Trebicka J, Jochem K, Zhou Q, et al. (2006) Defective RhoA/Rho-kinase signaling contributes to vascular hypocontractility and vasodilation in cirrhotic rats. Gastroenterology 130: 838–854.
[20]
Zhou Q, Hennenberg M, Trebicka J, Jochem K, Leifeld L, et al. (2006) Intrahepatic upregulation of RhoA and Rho-kinase signalling contributes to increased hepatic vascular resistance in rats with secondary biliary cirrhosis. Gut 55: 1296–1305.
[21]
Yang YY, Lin HC, Huang YT, Lee TY, Hou MC, et al. (2006) Roles of anandamide in the hepatic microcirculation in cirrhotic rats. Am J Physiol Gastrointest Liver Physiol 290: G328–G334.
[22]
Okumura H, Aramaki T, Katsuta Y, Satomura K, Akaike M, et al. (1991) Reduction in hepatic venous pressure gradient as a consequence of volume contraction due to chronic administration of spironolactone in patients with cirrhosis and no ascites. Am J Gastroenterol 86: 46–52.
[23]
Nevens F, Lijnen P, VanBilloen H, Fevery J (1996) The effect of long-term treatment with spironolactone on variceal pressure in patients with portal hypertension without ascites. Hepatology 23: 1047–1052.
[24]
Rombouts K, Niki T, Wielant A, Hellemans K, Schuppan D, et al. (2001) Effect of aldosterone on collagen steady state levels in primary and subcultured rat hepatic stellate cells. J Hepatol 34: 230–238.
[25]
Caligiuri A, De Franco RM, Romanelli RG, Gentilini A, Meucci M, et al. (2003) Antifibrogenic effects of canrenone, an antialdosteronic drug, on human hepatic stellate cells. Gatoroenterology 124: 504–520.
[26]
Yu J, Wu CW, Hui AY, Cheng AS, Go MY, et al. (2008) Elucidation of the role of COX-2 in liver fibrogenesis using transgenic mice. Biochem Biophys Res Commun 372: 571–577.
[27]
Wack KE, Ross MA, Zegarra V, Sysko LR, Watins SC, et al. (2001) Sinusoidal ultrastructure evaluated during the revascularization of regenerating rat liver. Hepatology 33: 363–378.
[28]
Reynaert H, Thompson MG, Thomas T, Geerts A (2002) Hepatic stellate cells: role in microcirculation and pathophysiology of portal hypertension. Gut 50: 571–581.
[29]
Friedman SL (2008) Mechanisms of hepatic fibrogenesis. Gastroenterology 134: 1655–1669.
[30]
Lee JS, Semela D, Iredale J, Shah VH (2007) Sinusoidal remodeling and angiogenesis: a new function for the liver-specific pericyte? Hepatology 45: 817–825.
[31]
Yang X, Li X, Wu P, Meng Y, Li S, et al. (2000) CYP11B2 expression in rat liver and the effect of spironolactone on hepatic fibrogenesis. Horm Res 53: 288–293.
[32]
Wada T, Kenmochi H, Miyashita Y, Sasaki M, Ojima M, et al. (2010) Spironolactone improves glucose and lipid metabolism by ameliorating hepatic steatosis and inflammation and suppressing enhanced gluconeogenesis induced by high-fat and high-fructose diet. Endocrinology 151(5): 2040–2049.
[33]
Rajagopalan S, Duquaine D, King S, Pitt B, Patel P (2002) Mineralocorticoid receptor antagonism in experimental atherosclerosis. Circulation 105: 2212–2216.
[34]
Leopold JA, Dam A, Maron BA, Scribner AW, Liao R, et al. (2007) Aldosterone impairs vascular reactivity by decreasing glucose-6-phosphate dehydrogenase activ-ity. Nat Med 13: 189–197.
[35]
Miyata K, Rahman M, Shokoji T, Nagai Y, Zhang GX, et al. (2005) Aldosterone stimulates reactive oxygen species production through activation of NADPH oxidase in rat mesangial cells. J Am Soc Nephrol 16: 2906–2912.
[36]
Fujisawa G, Okada K, Muto S, Fujita N, Itabashi N, et al. (2004) Spironolactone prevents early renal injury in streptozotocin-induced diabetic rats. Kidney Int 66: 1493–1502.
[37]
Sun Y, Zhang J, Lu L, Chen SS, Quinn MT, et al. (2002) Aldosterone-induced inflammation in the rat heart: role of oxidative stress. Am J Pathol 161: 1773–1781.
[38]
Rikitake Y, Liao JK (2005) Rho GTPases, statins, and nitric oxide. Circ Res 97: 1232–1235.
[39]
Ming XF, Viswambharan H, Barandier C, Ruffieux J, Kaibuchi K, et al. (2002) Rho GTPase/Rho kinase negatively regulates endothelial nitric oxide synthase phosphorylation through the inhibition of protein kinase B/Akt in human endothelial cells. Mol Cell Biol 22: 8467–8477.
[40]
Murthy KS, Zhou H, Grider JR, Makhlouf GM (2003) Inhibition of sustained mooth muscle contraction by PKA and PKG preferentially mediated by phosphorylation of RhoA. Am J Physiol Gastrointest Liver Physiol 284: G1006–G1016.
[41]
Wang Y, Gao J, Zhang D, Zhang J, Ma J, et al. (2010) New insights into the antifibrotic effects of sorafenib on hepatic stellate cells and liver fibrosis. J Hepatol 53(1): 132–144.
[42]
Hennenberg M, Trebicka J, Kohistani Z, Stark C, Nischalke HD, et al. (2011) Hepatic and HSC-specific sorafenib effects in rats with established secondary biliary cirrhosis. Lab Invest 91(2): 241–251.
[43]
Matei V, Rodríguez-Vilarrupla A, Deulofeu R, García-Calderó H, Fernández M, et al. (2008) Three-day tetrahydrobiopterin therapy increases in vivo hepatic NOS activity and reduces portal pressure in CCl4 cirrhotic rats. J Hepatol 49(2): 192–197.
[44]
Lavi？a B, Gracia-Sancho J, Rodríguez-Vilarrupla A, Chu Y, Heistad DD, et al. (2009) Superoxide dismutase gene transfer reduces portal pressure in CCl4 cirrhotic rats with portal hypertension. Gut 58(1): 118–125.
