Gene transfer of the neuronal NO synthase isoform to cirrhotic rat liver ameliorates portal hypertension
Qing Yu, … , Samuel E. George, Don C. Rockey
Qing Yu, … , Samuel E. George, Don C. Rockey
Published March 15, 2000
Citation Information: J Clin Invest. 2000;105(6):741-748. https://doi.org/10.1172/JCI7997.
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Article

Gene transfer of the neuronal NO synthase isoform to cirrhotic rat liver ameliorates portal hypertension

Abstract

Reduced production of nitric oxide (NO) in the cirrhotic liver results from a defect in hepatic endothelial cell nitric oxide synthase (ecNOS) and appears to contribute to the high intrahepatic resistance and portal hypertension typical of cirrhosis. Therefore, we postulated that targeting a heterologous NOS isoform to sinusoidal endothelial cells or other perisinusoidal cells, such as hepatic stellate cells, would counter the defect in NO production and reduce resistance to blood flow. Recombinant adenovirus (Ad) carrying the neuronal NOS gene (nNOS) targeted liver sinusoidal endothelial cells, stellate cells, and hepatocytes more efficiently than the corresponding cells in cirrhotic livers, but transduction rates were substantial even in cirrhotic animals. Expression of nNOS in each liver cell type, whether from normal or injured liver, caused increased NO production and inhibited endothelin-1–induced contractility of perisinusoidal stellate cells. Finally, in 2 different in vivo models of cirrhosis and portal hypertension, transduction of livers with recombinant Ad.nNOS significantly reduced intrahepatic resistance and portal pressure. The data highlight the feasibility of gene transfer to diseased liver and hepatic cells and demonstrate the potential of a novel therapy for portal hypertension caused by cirrhosis.

Authors

Qing Yu, Rong Shao, Hu Sheng Qian, Samuel E. George, Don C. Rockey

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Ad-mediated nNOS gene delivery and activity of nNOS. Parenchymal and non...
Ad-mediated nNOS gene delivery and activity of nNOS. Parenchymal and nonparenchymal cells were prepared as in Methods. After isolation, cells were grown for 48–72 hours under standard conditions and then transduced with Ad.β-gal or Ad.nNOS as in Methods. In a and b, cells were transduced with Ad.β-gal or Ad.nNOS, respectively, and stained for NADPH diaphorase. In c, the transduction efficiency after exposure to Ad.nNOS was determined by cell counting as in Methods (Ad.β-gal is used as an additional control). In d, cell lysates (25 μg total protein) were immunoblotted for nNOS. In e, 24 hours after transduction, culture supernatants were collected and nitrite detected. AP < 0.01 compared with control or Ad.β-gal for each cell type (n = 3). H, hepatocyte; SEC, sinusoidal endothelial cells; HSC, hepatic stellate cells.