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

Figure 3 Hepatic soluble epoxide hydrolase status and epoxyeicosatrienoic acids level in choline-deficient, L-amino acid-defined, high-fat diet-fed rats. A: Immunofluorescence with soluble epoxide hydrolase (sEH) in liver tissues. Scale bar: 50 μm. Nuclei were stained by 4’,6-diamidino-2-phenylindole; B: Quantification of sEH positive area in liver tissues; C: SEH activity in liver tissues; D: Hepatic levels of 11,12-epoxyeicosatrienoic acids (EET) and 14,15-EET; E and F: Protein (E) and message RNA (mRNA) (F) levels of angiotensin II receptor type 1 (AT1R) in liver tissues; G: Pearson’s correlation between sEH expression and AT1R protein level in all of experimental rats; H and I: Protein (H) and mRNA (I) levels of nicotinamide adenine dinucleotide phosphate hydrogen oxidase 4 (NOX4) in liver tissues; J: Pearson’s correlation between sEH expression and NOX4 protein level in all of experimental rats. 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, F, I). Data are the mean ± SD (n = 6, B-F, H, I). ^aP < 0.05. ^bP < 0.01. Significant difference between groups determined by Student’s t-test. mRNA: Message RNA; DAPI: 4’,6-diamidino-2-phenylindole; CSANFD: Choline-supplemented L-amino acid-defined, normal-fat diet; CDAHFD: Choline-deficient, L-amino acid-defined, high-fat diet; sEH: Soluble epoxide hydrolase; 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.