Modulation of Cardiovascular Function in Primary Hypertension in Rat by SKA-31, an Activator of KCa2.x and KCa3.1 Channels
Abstract
:1. Introduction
2. Results
2.1. General
2.2. Influence of SKA-31 on BP and HR of SHR and WKY Rats
2.3. Influence of Endothelial Physical Disruption, INDO and l-NAME on SKA-31-Induced Relaxation
2.4. Influence of KCa, KIR2.1, and Na+/K+-ATPase Inhibitors on EDH-Type Relaxation Induced by SKA-31
2.5. Vasodilatory Effects of SKA-31, NS309, and Acetylcholine in sMAs
3. Discussion
3.1. Vascular Changes Related to Hypertension
3.2. Influence of SKA-31 on BP and HR
3.3. Influence of SKA-31 on Vasodilatory Effects in sMAs
3.4. Involvement of KCa2.3 and KCa3.1 Channels in EDH-Type Relaxation Induced by SKA-31
3.5. Downstream Signaling Pathway in EDH-Type Relaxation Induced by SKA-31
3.6. Influence of SKA-31 on Potentiation of Ach-Induced EDH in sMAs
3.7. Limitations
4. Materials and methods
4.1. Animals
4.2. Measurements of Blood Pressure (BP)
4.3. In Vivo Effects of SKA-31 in SHR and WKY Rats
4.4. Vessel Preparation
4.5. Concentration-Response Curves
4.6. Histopathological Examination
4.6.1. Width of Tunica Media in Mesenteric Arteries
4.6.2. Immunohistochemistry
4.7. Real-Time qPCR
4.8. Drugs
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
Ach | acetylcholine |
AMPK | AMP-activated protein kinase |
CRCs | concentration-response curves |
COX | cyclooxygenase |
DBP | diastolic blood pressure |
DMSO | dimethyl sulfoxide |
EDH | endothelium-derived hyperpolarization |
HR | heart rate |
i.p. | intraperitoneal |
i.v. | intravenous |
INDO | indomethacin |
KCa | calcium-activated potassium channel |
KCa2.x | small conductance calcium-activated potassium channels |
KCa2.3 | type 3 endothelial small conductance calcium-activated potassium channels |
KCa3.1 | intermediate conductance calcium-activated potassium channel |
KIR | inward-rectifier potassium ion channel |
l-NAME | Nω-nitro-l-arginine methyl ester |
NOS | nitric oxide synthase |
PGIS | prostacyclin I2 synthase |
Phe | phenylephrine |
SBP | systolic blood pressure |
SKA-31 | (naphtha(1–d)thiazol-2-ylamine) |
SIRT1 | silent information regulator T1, sirtuin-1 |
sMAs | small mesenteric arteries |
SHR | spontaneously hypertensive rats |
WKY | Wistar-Kyoto |
SHRSP | stroke-prone SHR |
vWF | von Willebrand factor |
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Group | n | Before SKA-31 (1 mg/kg)/veh | Before SKA-31 (3 mg/kg)/veh | Before SKA-31 (10 mg/kg)/veh | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
DBP | SBP | MAP | HR | DBP | SBP | MAP | HR | DBP | SBP | MAP | HR | ||
WKY+SKA-31 | 4 | 40 ± 2 | 82 ± 4 | 54 ± 3 | 258 ± 7 | 43 ± 2 | 86 ± 5 | 57 ± 3 | 263 ± 11 | 45 ± 2 | 95 ± 5 | 62 ± 3 | 262 ± 6 |
WKY+veh | 4 | 40 ± 4 | 81 ± 6 | 54 ± 5 | 251 ± 19 | 44 ± 3 | 86 ± 4 | 58 ± 3 | 235 ± 10 | 44 ± 2 | 87 ± 2 | 58 ± 2 | 235 ± 12 |
SHR+SKA-31 | 4 | 52 ± 4 | 113 ± 7 ** | 72 ± 4 * | 320 ± 27 | 55 ± 3 | 120 ± 5 ** | 77 ± 4 * | 347 ± 33 | 56 ± 4 | 121 ± 1 | 78 ± 3 * | 371 ± 31 * |
SHR+veh | 4 | 52 ± 3 | 106 ± 3 * | 70 ± 2 * | 287 ± 21 | 51 ± 3 | 109 ± 3 * | 70 ± 3 | 294 ± 26 | 54 ± 4 | 115 ± 4 | 74 ± 4 * | 320 ± 23 |
Group | Concentration in (μM) | ENDO | WKY | SHR | ||||||
---|---|---|---|---|---|---|---|---|---|---|
n | Tension (mN) | pEC25 | Rmax (%) | n | Tension (mN) | pEC25 | Rmax (%) | |||
SKA-31 | 0.01–10 | + | 12 | 10.6 ±1.2 | 6.8 ± 0.10 | 92.8 ± 2.3 | 13 | 10.7 ± 1.1 | 6.1 ± 0.07 &&& | 75.8 ± 6.1 & |
SKA-31 | 0.01–10 | − | 11 | 9.6 ± 1.4 | 5.2 ± 0.06 *** | 33.2 ± 2.1 *** | 4 | 9.0 ± 1.3 | N.D. | 19.6 ± 7.1 ***,& |
+INDO | 10 | + | 5 | 9.5 ± 1.6 | 6.1 ± 0.07 *** | 91.4 ± 6.5 | 6 | 10.9 ± 1.5 | 5.7 ± 0.06 * | 81.0 ± 3.2 |
+l-NAME | 100 | + | 5 | 10.2 ± 1.5 | 5.7 ± 0.10 *** | 68.2 ± 5.4 *** | 5 | 11.0 ± 1.0 | 5.6 ± 0.13 ** | 68.1 ± 5.6 |
+l-NAME+INDO (EDH) | 100 + 10 | + | 14 | 10.7 ± 0.8 | 5.6 ± 0.07 *** | 64.9 ± 7.4 *** | 15 | 11.4 ± 1.0 | 5.6 ± 0.10 ** | 63.1 ± 6.6 |
+UCL1684 | 0.1 | + | 9 | 11.6 ± 1.4 | 5.3 ± 0.06 # | 43.4 ± 4.3 | 8 | 10.7 ± 1.5 | 5.3 ± 0.08 # | 46.5 ± 4.0 |
+TRAM-34 | 10 | + | 6 | 9.3 ± 0.8 | 5.2 ± 0.10 ## | 36.4 ± 5.8 # | 11 | 9.1 ± 1.6 | 5.2 ± 0.07 ## | 32.2 ± 6.1 ## |
+Ba2+ | 30 | + | 9 | 8.9 ± 1.6 | 5.0 ± 0.20 ## | 25.7 ± 3.1### | 10 | 10.7 ± 1.1 | 5.1 ± 0.10 ## | 38.2 ± 3.9 ###,& |
+OUABAIN | 100 | + | 10 | 9.4 ± 1.1 | 5.2 ± 0.06 # | 37.3 ± 3.2 ## | 11 | 10.6 ± 1.2 | N.D. | 13.6 ± 3.3 ###,&& |
Primer | Sequence | References |
---|---|---|
KCa2.3 | Forward: 5′-CGCCTTCAGAATAGAGTT-3′ Reverse: 5′-GAGTGTGCATTGTATTGG-3′ | [61] |
KCa3.1 | Forward: 5′-CTGAGATGTTGTGGTTCCT-3′ Reverse: 5′-CAGTGGACAGCGTGATTA-3′ | [61] |
eNOS | Forward: 5′-AGCATGAGGCCTTGGTATTG-3′ Reverse: 5′-CCCGACATTTCCATCAGC-3′ | [62] |
Na+/K+-ATPase | Forward: 5′-GAAGCTCATCATCAGGCGACG-3′ Reverse: 5′-CCAGGGTAGAGTTCCGAGCTC-3′ | [63] |
KIR2.1 | Forward: 5′-CACGGGGATCTGGATGCTTCTAAA-3′ Reverse: 5′-AGCAATCGGGCACTCGTCTGTAAC-3′ | [63] |
PGIS | Forward: 5′-TTTTACAGATGACCGCACTCC-3′ Reverse: 5′-GAAATGAGTCAGCAGCAGGAC-3′ | [64] |
GAPDH | Forward: 5′-TGACTCTACCCACGGCAAGTT-3′ Reverse: 5′-TGATGGGTTTCCCGTTGATGA-3′ | [61] |
β-actin | Forward: 5′-GGGAAATCGTGCGTGACATT-3′ Reverse: 5′-GCGGCAGTGGCCATCTC-3′ | [61] |
cyclophilin A | Forward: 5′-TGTCTCTTTTCGCCGCTTGCTG-3′ Reverse: 5′-CACCACCCTGGCACATGAATCC-3′ | [65] |
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Kloza, M.; Baranowska-Kuczko, M.; Toczek, M.; Kusaczuk, M.; Sadowska, O.; Kasacka, I.; Kozłowska, H. Modulation of Cardiovascular Function in Primary Hypertension in Rat by SKA-31, an Activator of KCa2.x and KCa3.1 Channels. Int. J. Mol. Sci. 2019, 20, 4118. https://doi.org/10.3390/ijms20174118
Kloza M, Baranowska-Kuczko M, Toczek M, Kusaczuk M, Sadowska O, Kasacka I, Kozłowska H. Modulation of Cardiovascular Function in Primary Hypertension in Rat by SKA-31, an Activator of KCa2.x and KCa3.1 Channels. International Journal of Molecular Sciences. 2019; 20(17):4118. https://doi.org/10.3390/ijms20174118
Chicago/Turabian StyleKloza, Monika, Marta Baranowska-Kuczko, Marek Toczek, Magdalena Kusaczuk, Olga Sadowska, Irena Kasacka, and Hanna Kozłowska. 2019. "Modulation of Cardiovascular Function in Primary Hypertension in Rat by SKA-31, an Activator of KCa2.x and KCa3.1 Channels" International Journal of Molecular Sciences 20, no. 17: 4118. https://doi.org/10.3390/ijms20174118