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

Figure 6 Hepatic fibrosis development in choline-deficient, L-amino acid-defined, high-fat diet-fed rats. A: Sirius-red staining and immunofluorescence with alpha-smooth muscle actin (α-SMA) in liver tissues. Scale bar: 50 μm. Nuclei were stained by 4’,6-diamidino-2-phenylindole; B: Quantification of Sirius-red-stained fibrotic area; C: Hepatic content of hydroxyproline; D: Quantification of α-SMA-positive hepatic stellate cells; E: Hepatic message RNA (mRNA) levels of profibrogenic markers (Acta2, Col1a1, Tgfb1 and Ctgf). 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-deficient, L-amino acid-defined, high-fat die-fed groups + vehicle (saline)-treated group (B, D and E). Data are the mean ± SD (n = 6, B-E). ^aP < 0.05. ^bP < 0.01. Significant difference between groups determined by Student’s t-test. CSANFD: Choline-supplemented L-amino acid-defined, normal-fat diet; CDAHFD: Choline-deficient, L-amino acid-defined, high-fat diet; mRNA: Message RNA; DAPI: 4’,6-diamidino-2-phenylindole; α-SMA: Alpha-smooth muscle actin; 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.