Investigation of Genes and Proteins Expression Associating Serotonin Signaling Pathway in Lung and Pulmonary Artery Tissues of Dogs with Pulmonary Hypertension Secondary to Degenerative Mitral Valve Disease: The Preliminary Study

SIMPLE SUMMARY: Pulmonary hypertension is an unusual elevation of the blood pressure in pulmonary circulation. In dogs, pulmonary hypertension is commonly occurred as a complication of progressive degenerative mitral valve disease especially in senior small-breed dogs. Previous studies revealed that the serotonin signaling pathway is associated with the pathogenesis of pulmonary hypertension. However, research in dogs was scarce. Therefore, the present study aimed to illustrate the understanding of this point by assessing the expression of the targeted genes and proteins related to the serotonin pathway in lung tissues and pulmonary arteries of dogs. Our results showed that the pattern of gene and protein expression was different in canine lung and pulmonary arteries. The targeted proteins in pulmonary artery tissues of the degenerative mitral valve disease dogs with and without pulmonary hypertension tended to be upregulated. In addition, the expression of these protein was increased in the pneumocytes of the alveolar walls, pulmonary alveolar macrophages, and bronchial epithelial cells. A correlation between the targeted gene and protein expression and the echocardiographic data was also founded. Overall results pointed out that the serotonin pathway in lung and pulmonary artery tissues might have different roles in degenerative mitral disease with and without pulmonary hypertension. ABSTRACT: Pulmonary hypertension (PH) is defined as an increase in pulmonary vascular pressure. It is one of the most common complications that occur as a result of degenerative mitral valve disease (DMVD) in dogs. Serotonin (5-HT) can trigger the development of PH. Accordingly, this study investigated the changes in the expression of genes and proteins associated with local 5-HT signaling in the lungs and pulmonary arteries (PA) of dogs with PH secondary to DMVD. Lung and PA tissue samples were collected from the cadavers of fourteen small-breed dogs and divided into normal (n = 4), DMVD (n = 5) and DMVD with PH (n = 5) groups. Gene expression (tph1, slc6a4 and htr2a) was analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The expression of proteins (TPH-1, SERT, 5-HTR2A, ERK1/2 and pERK1/2) was examined by Western blot analysis and immunohistochemical staining. The results showed that the expression of genes and proteins evaluated by qRT-PCR and Western blot analysis in lung and PA tissues did not differ among groups. However, the expression of proteins related to 5-HT signaling tended to be upregulated in PA tissues from DMVD dogs with and without PH. Immunohistochemical examination revealed the overexpression of these proteins in the DMVD and DMVD with PH groups in lung tissue. These findings suggest a local effect of 5-HT signaling in DMVD dogs with and without PH.