Dual therapy with vildagliptin and sacubitril/valsartan alleviates portal hypertension and inhibits soluble epoxide hydrolase in cirrhotic rats

Figure 2 Intrahepatic vasoconstriction and vascular resistance in choline-deficient, L-amino acid-defined, high-fat diet-fed rats. A: Hepatic level of endothelin-1; B: Hepatic message RNA (mRNA) levels of cystathionine γ-lyase and dimethylarginine dimethyl amino-hydrolase 1; C: Western blotting for the phosphorylation of endothelial nitric oxide (NO) synthase (eNOS), inducible NO synthase (iNOS) and moesin in liver tissues; D-F: Quantitative phosphorylation rate of phospho-eNOS/total eNOS (D), phospho-iNOS/total iNOS (E) and phospho-moesin/total moesin (F). Actin was used as the loading control; G: Hepatic mRNA levels of guanosine triphosphate cyclohydrolase 1; H: Hepatic level of NO (quantified as the total amount of nitrate and nitrite); I: Hepatic level of cyclic guanosine monophosphate. The mRNA expression levels were measured by quantitative real-time polymerase chain reaction, and glyceraldehyde-3-phosphate dehydrogenase was used as internal control. Quantitative values are indicated as fold changes to the values of choline-supplemented L-amino acid-defined, normal-fat diet-fed group + vehicle (saline)-treated group (B, D-G). Data are the mean ± SD (A, B, G-I: n = 6; D-F: n = 3). ^aP < 0.05. ^bP < 0.01. Significant difference between groups determined by Student’s t-test (A, B, G-I) or Mann-Whitney U test (D-F). mRNA: Message RNA; eNOS: Endothelial nitric oxide synthase; iNOS: Inducible nitric oxide synthase; p-eNOS: Phospho-endothelial nitric oxide synthase; p-iNOS: Phospho-inducible nitric oxide synthase; Veh: Vehicle (saline)-treated groups; DP4i: Dipeptidyl peptidase-4 inhibitor (vildagliptin)-treated group; ARNI group: Angiotensin receptor-neprilysin inhibitor (sacubitril/valsartan)-treated group; CS: Choline-supplemented L-amino acid-defined, normal-fat diet-fed group; CD: Choline-deficient, L-amino acid-defined, high-fat die-fed groups. “Both group” defined dipeptidyl peptidase-4 inhibitor and angiotensin receptor-neprilysin inhibitor-treated group.