title: Role of estrogen receptor β selective agonist in ameliorating portal hypertension in rats with CCl4-induced liver cirrhosis.
abstract: OBJECTIVE
  To investigate the role of diarylpropionitrile (DPN), a selective agonist of estrogen receptor β (ERβ), in liver cirrhosis with portal hypertension (PHT) and isolated hepatic stellate cells (HSCs).
  
  
  METHODS
  Female Sprague-Dawley rats were ovariectomized (OVX), and liver cirrhosis with PHT was induced by CCl4 injection. DPN and PHTPP, the selective ERβ agonist and antagonist, were used as drug interventions. Liver fibrosis was assessed by hematoxylin and eosin (HE) and Masson's trichrome staining and by analyzing smooth muscle actin expression. Hemodynamic parameters were determined in vivo using colored microspheres technique. Protein expression and phosphorylation were determined by immunohistochemical staining and Western blot analysis. Messenger RNA levels were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Collagen gel contraction assay was performed using gel lattices containing HSCs treated with DPN, PHTPP, or Y-27632 prior to ET-1 addition.
  
  
  RESULTS
  Treatment with DPN in vivo greatly lowered portal pressure and improved hemodynamic parameters without affecting mean arterial pressure, which was associated with the attenuation of liver fibrosis and intrahepatic vascular resistance (IHVR). In CCl4-treated rat livers, DPN significantly decreased the expression of RhoA and ROCK II, and even suppressed ROCK II activity. Moreover, DPN remarkedly increased the levels of endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS, and promoted the activities of protein kinase G (PKG), which is an NO effector in the liver. Furthermore, DPN reduced the contractility of activated HSCs in the 3-dimensional stress-relaxed collagen lattices, and decreased the ROCK II activity in activated HSCs. Finally, in vivo/in vitro experiments demonstrated that MLC activity was inhibited by DPN.
  
  
  CONCLUSIONS
  For OVX rats with liver cirrhosis, DPN suppressed liver RhoA/ROCK signal, facilitated NO/PKG pathways, and decreased IHVR, giving rise to reduced portal pressure. Therefore, DPN represents a relevant treatment choice against PHT in cirrhotic patients, especially postmenopausal women.
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Cheng-Gang
authors.initials: CG
authors.lastname: Zhang
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Wen-Sheng
authors.initials: WS
authors.lastname: Deng
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Duan
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Cheng-Gang Zhang, Bin Zhang, Wen-Sheng Deng, Ming Duan, Wei Chen, Zhi-Yong Wu, Department of Gastrointestinal Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Wu
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002251:Carbon Tetrachloride; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000188:drug therapy* / Q000209:etiology / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D047629:Estrogen Receptor beta / Q000819:agonists* / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D055166:Hepatic Stellate Cells / Q000187:drug effects / Q000378:metabolism; D006975:Hypertension, Portal / Q000209:etiology / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control*; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism; D008297:Male; D018994:Myosin Light Chains / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology*; D010052:Ovariectomy; D010766:Phosphorylation; D017082:Portal Pressure / Q000187:drug effects*; D011422:Propionates / Q000494:pharmacology*; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D017207:Rats, Sprague-Dawley; D020845:Selective Estrogen Receptor Modulators / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D014655:Vascular Resistance / Q000187:drug effects; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D000199:Actins; D002251:Carbon Tetrachloride; D017869:Cyclic GMP-Dependent Protein Kinases; D047629:Estrogen Receptor beta; D004967:Estrogens; D018994:Myosin Light Chains; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D009570:Nitriles; D011422:Propionates; D011720:Pyrazoles; D011743:Pyrimidines; D020845:Selective Estrogen Receptor Modulators; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; C486182:2,3-bis(4-hydroxyphenyl)-propionitrile; C556295:4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo(1,5-a)pyrimidin-3-yl)phenol; C498862:Acta2 protein, rat; C496316:Nos3 protein, rat; C580818:ROCK2 protein, rat
keywords: Estrogen receptor; Hepatic stellate cells; Nitric oxide; Portal hypertension; Rho-kinase signaling