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
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Animals
2.2. Investigation of Genes Related to 5-HT Signaling Pathway
2.3. Investigation of the Downstream Effectors in 5-HT Signaling Pathway by Western Blot Analysis
2.4. Investigation of the Downstream Effectors in 5-HT Signaling Pathway by Immunohistochemistry
2.5. Statistical Analysis
3. Results
3.1. Animals
3.2. Investigation of Genes Associating 5-HT Signaling Pathway
3.3. Investigation of Proteins Associating 5-HT Signaling Pathway
3.3.1. Western Blot Analysis
3.3.2. Immunohistochemical Study
3.4. The Correlation between the Relative Expression and Echocardiographic Parameters
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Genes | Accession No. | Forward Primer (5′ to 3′) | Reverse Primer (5′ to 3′) | Product Size (bp) | Tm (°C) | Reference of Primers |
---|---|---|---|---|---|---|
tph1 | NM_001197191.1 | CTGTGGAGTTTGGTCTCTGTAAG | TGTGATGAGACACTCCTGTTTG | 158 | 82.77 | This study |
slc6a4 | NM_001110771.1 | GGCTGAGATGAGGAACGAAG | TTGGACCAGATGTGTGGAAA | 222 | 84.43 | This study |
htr2a | NM_001005869.1 | TCTTTCAGCTTCCTCCCTCA | TCCTCGTTGCAGGACTCTTT | 227 | 84.70 | This study |
RPS19 | XM_005616513.3 | CCTTCCTCAAAAAGTCTGGG | GTTCTCATCGTAGGGAGCAAG | 95 | 80.69 | Brinkhof et al. [39] |
RPL32 | XM_022406256.1 | TGGTTACAGGAGCAACAAGAAA | GCACATCAGCAGCACTTCA | 100 | 81.54 | Peters, Peeters, Helps and Day [37] |
Normal (n = 4) | DMVD (n = 5) | DMVD with PH (n = 5) | p-Value | |
---|---|---|---|---|
Sex (male/female) | 3/1 | 1/4 | 3/2 | - |
Age (years) | 8 (7.8–10.0) | 15 (14–15) | 14 (13–15) | |
Weight (kg) | 5.6 (5.28–6.93) | 3.92 (3.7–4.14) | 4.3 (2.8–6.9) | |
Breed | Shih-Tzu (n = 2) Cocker Spaniel (n = 1) Mixed breed (n = 1) | Poodle (n = 3) Shih-Tzu (n = 1) Pomeranian (n = 1) | Chihuahua (n = 2) Poodle (n = 2) Pomeranian (n = 1) | - |
Stage (C/D) | - | 2/3 | 1/4 | - |
Medication | - | ACEIs (n = 5) Furosemide (n = 5) Pimobendane (n = 5) Spironolactone (n = 2) Moduretic (n = 1) | ACEIs (n = 5) Furosemide (n = 5) Pimobendane (n = 5) Spironolactone (n = 2) Moduretic (n = 2) Sildenafil (n = 1) | - |
DMVD (n = 5) | DMVD with PH (n = 5) | p-Value | |
---|---|---|---|
LA index (cm/kg) | 1.67 ± 0.28 | 1.83 ± 0.72 | 0.642 |
Ao index (cm/kg) | 1.01 ± 0.42 | 1.02 ± 0.28 | 0.952 |
LA/Ao | 1.80 ± 0.56 | 1.75 ± 0.38 | 0.862 |
IVSdN index (cm/kg) | 0.59 ± 0.13 | 0.56 ± 0.14 | 0.663 |
LVIDdN index (cm/kg) | 2.84 ± 0.87 | 1.66 ± 0.62 | 0.038 |
LVPWdN index (cm/kg) | 0.55 ± 0.24 | 0.48 ± 0.17 | 0.621 |
IVSsN index (cm/kg) | 0.92 ± 0.30 | 0.84 ± 0.29 | 0.707 |
LVIDsN index (cm/kg) | 1.59 ± 0.50 | 0.72 ± 0.30 | 0.010 |
LVPWsN index (cm/kg) | 0.85 ± 0.33 | 0.71 ± 0.10 | 0.407 |
%FS | 42.14 ± 5.82 | 55.16 ± 7.91 | 0.018 |
PAP (mmHg) | - | 72.03 ± 25.96 | - |
Gene | Normal (n = 4) | DMVD (n = 5) | DMVD with PH (n = 5) | p-Value |
---|---|---|---|---|
Lung tissues (×10−3) | ||||
tph1 | 0.61 (0.04–3.29) | 0.24 (0.06–4.21) | 0.13 (0.07–0.53) | 0.680 |
slc6a4 | 3.32 (0.19–6.23) | 1.11 (0.58–3.03) | 0.27 (0.13–1.03) | 0.081 |
htr2a | 0.33 (0.23–0.68) | 0.10 (0.09–0.55) | 0.33 (0.29–0.34) | 0.154 |
PA tissues (×10−3) | ||||
tph1 | 0.20 (0.04–3.89) | 1.89 (0.58–6.58) | 1.01 (0.45–1.71) | 0.117 |
slc6a4 | 1.60 (0.08–5.89) | 11.03 (10.66–11.40) | 0.74 (0.12–14.94) | 0.252 |
htr2a | 14.46 (0.85–20.05) | 2.00 (0.43–3.22) | 5.79 (0.41–19.19) | 0.270 |
Proteins | Normal (n = 4) | DMVD (n = 5) | DMVD with PH (n = 5) | p-Value |
---|---|---|---|---|
Lung tissues | ||||
TPH-1 | 2.33 (0.58–4.09) | 1.59 (0.52–5.43) | 1.56 (0.68–5.08) | 0.362 |
SERT | 1.14 (0.09–3.10) | 0.46 (0.04–1.00) | 1.49 (0.24–1.69) | 0.373 |
5-HTR2A | 4.40 (0.97–10.75) | 2.59 (1.12–9.46) | 3.15 (2.36–8.88) | 0.656 |
ERK1/2 | 1.74 (0.68–6.37) | 1.41 (0.72–5.38) | 2.16 (1.04–4.29) | 0.680 |
pERK1/2 | 0.72 (0.13–2.01) | 1.02 (0.35–4.09) | 0.21 (0.07–2.59) | 0.284 |
PA tissues | ||||
TPH-1 | 0.65 (0.49–1.12) | 0.81 (0.29–4.09) | 1.85 (0.51–2.95) | 0.401 |
SERT | 0.23 (0.14–0.39) | 0.14 (0.04–0.63) | 0.40 (0.04–0.74) | 0.761 |
5-HTR2A | 0.68 (0.44–1.00) | 0.99 (0.37–4.67) | 1.58 (0.50–2.57) | 0.350 |
ERK1/2 | 0.86 (0.73–1.04) | 0.81 (0.54–4.23) | 1.04 (0.48–4.15) | 0.831 |
pERK1/2 | 0.61 (0.35–0.89) | 0.45 (0.11–2.26) | 0.56 (0.10–2.87) | 0.990 |
Proteins | Cells | Normal (n = 4) | DMVD (n = 5) | DMVD with PH (n = 5) | p-Value |
---|---|---|---|---|---|
TPH-1 | Pneumocytes | 1.63 ± 0.13 | 2.76 ± 0.57 a,b | 4.90 ± 0.99 a,c | <0.001 |
PAMs Bronchial epithelial cells | 0.28 ± 0.16 74.84 ± 2.82 | 0.65 ± 0.26 76.62 ± 5.33 | 0.52 ± 0.21 81.83 ± 8.98 | 0.074 0.273 | |
SERT | Pneumocytes | 1.28 ± 0.24 | 3.50 ± 1.03 a | 4.35 ± 1.11 a | <0.001 |
PAMs Bronchial epithelial cells | 0.29 ± 0.14 67.59 ± 3.12 | 1.63 ± 0.94 a 69.87 ± 5.09 | 1.23 ± 0.47 a 71.90 ± 4.37 | 0.028 0.371 | |
5-HTR2A | Pneumocytes | 3.89 ± 0.48 | 7.36 ± 1.03 a | 7.29 ± 1.24 a | <0.001 |
PAMs Bronchial epithelial cells | 1.67 ± 0.29 86.52 ± 3.90 | 3.62 ± 1.00 a 84.79 ± 7.24 | 3.73 ± 0.65 a 87.32 ± 6.24 | 0.005 0.804 | |
ERK1/2 | Pneumocytes | 0.81 ± 0.10 | 2.40 ± 0.25 a,b | 1.54 ± 0.41 a,c | <0.001 |
PAMs Bronchial epithelial cells | 0.24 ± 0.02 78.84 ± 6.55 | 0.78 ± 0.43 76.82 ± 3.51 | 0.41 ± 0.16 81.01 ± 6.95 | 0.059 0.540 | |
pERK1/2 | Pneumocytes | 0.86 ± 0.18 | 1.63 ± 0.68 | 1.61 ± 0.43 | 0.243 |
PAMs Bronchial epithelial cells | 0.00 ± 0.00 0.00 ± 0.00 | 0.49 ± 0.30 26.16 ± 4.39 a,b | 0.53 ± 0.28 92.39 ± 7.19 a,c | 0.108 < 0.001 |
Organ or Cell | Genes or Proteins | Echo Parameters | r | p-Value |
---|---|---|---|---|
Lung | tph1 | LA/Ao | 0.648 | 0.043 |
pERK1/2 | LA/Ao | 0.758 | 0.011 | |
pERK1/2 | LVIDsN | 0.721 | 0.019 | |
PA | slc6a4 | LVIDdN | 0.857 | 0.014 |
slc6a4 | LVIDsN | 0.857 | 0.014 | |
5-HTR2A | PAP | −0.943 | 0.005 | |
Pneumocyte | TPH-1 | %FS | 0.705 | 0.023 |
TPH-1 | PAP | 0.829 | 0.042 | |
5-HTR2A | IVSdN | −0.693 | 0.026 | |
pERK1/2 | LVPWsN | 0.638 | 0.047 | |
PAM | SERT | PAP | 0.829 | 0.042 |
ERK1/2 | LVPWsN | 0.648 | 0.043 | |
pERK1/2 | LA | 0.721 | 0.019 | |
pERK1/2 | LA/Ao | 0.685 | 0.029 |
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Tangmahakul, N.; Sakarin, S.; Techangamsuwan, S.; Rungsipipat, A.; Surachetpong, S.D. 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. Vet. Sci. 2022, 9, 530. https://doi.org/10.3390/vetsci9100530
Tangmahakul N, Sakarin S, Techangamsuwan S, Rungsipipat A, Surachetpong SD. 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. Veterinary Sciences. 2022; 9(10):530. https://doi.org/10.3390/vetsci9100530
Chicago/Turabian StyleTangmahakul, Nattawan, Siriwan Sakarin, Somporn Techangamsuwan, Anudep Rungsipipat, and Sirilak Disatian Surachetpong. 2022. "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" Veterinary Sciences 9, no. 10: 530. https://doi.org/10.3390/vetsci9100530