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

Figure 4 Sinusoidal capillarization and intrahepatic angiogenesis in choline-deficient, L-amino acid-defined, high-fat diet-fed rats. A: Immunofluorescence with cluster of differentiation 34 in liver tissues. Scale bar: 50 μm. Nuclei were stained by 4’,6-diamidino-2-phenylindole; B: Quantification of cluster of differentiation 34-positive sinusoidal capillarization; C: Hepatic message RNA (mRNA) levels of capillary markers (platelet endothelial cell adhesion molecule-1, endothelin-1 and laminin subunit beta-1); D: Hepatic mRNA levels of sinusoidal endothelial markers (lymphatic vessel endothelial hyaluronan receptor 1, plasmalemma vesicle-associated protein and stabilin-2; E: Hepatic mRNA levels of Hedgehog signaling-related genes (Shh, Gli2, Gli3, Sfrp1 and Opn); F: Hepatic mRNA levels of genes related to vascular injury (vascular cell adhesion molecule 1, intercellular adhesion molecule 1, and neurogenic locus notch homolog protein 1); G: Hepatic mRNA levels of proangiogenic factors (Vegfa, Ptgs2, Pigf and Vwf). 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-G). Data are the mean ± SD (n = 6, B-G). ^aP < 0.05; ^bP < 0.01. Significant difference between groups determined by Student’s t-test. CD34: Cluster of differentiation 34; 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; Pecam1: Platelet endothelial cell adhesion molecule-1; ET-1: Endothelin-1; Lamb1: Laminin subunit beta-1; Lyve1: Lymphatic vessel endothelial hyaluronan receptor 1; Plvap: Plasmalemma vesicle-associated protein; Stab2: Stabilin-2; Vcam1: Vascular cell adhesion molecule 1; Icam1: Intercellular adhesion molecule 1; Notch1: Neurogenic locus notch homolog protein 1; 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.