Molecular Mechanisms and Experimental Protocols in the Study of Vasorelaxant Activity in Aortic Smooth Muscle Cells
DOI:
https://doi.org/10.48048/tis.2026.11549Keywords:
Vasorelaxation, Calcium channels, SERCA, Ryanodine receptors, Na⁺/Ca²⁺ exchangerAbstract
Vasorelaxation is a key process in cardiovascular regulation and a promising target for treating hypertension and vascular dysfunctions. This study outlines a comprehensive experimental framework for evaluating vasorelaxant mechanisms in isolated rat aortic rings using isometric tension recording. A combination of pharmacological tools and ionic modulation was used to explore the roles of voltage-dependent calcium channels, GPCR signaling, intracellular calcium stores (SERCA and ryanodine receptors), ion exchangers (NCX, Na⁺/K⁺-ATPase) and endothelium-derived factors. Computational docking complemented experimental data by identifying ligand-binding sites on key ion channels. Findings revealed that vasorelaxation involves multifaceted modulation of calcium influx, receptor-mediated signaling, and intracellular calcium cycling, with both endothelium-dependent and -independent pathways contributing. This integrative approach provides novel insights into vascular smooth muscle regulation and supports the development of targeted cardiovascular therapies.
HIGHLIGHTS
- Isolated rat aortic rings were used to evaluate vasorelaxation via isometric tension recording.
- Pharmacological modulators revealed roles of VGCCs, GPCR pathways, SERCA, RyR, NCX, and Na⁺/K⁺-ATPase in vascular relaxation.
- Endothelium-dependent and -independent mechanisms both contributed to the vasorelaxant responses.
- Intracellular Ca²⁺ cycling and reduced Ca²⁺ influx were identified as key determinants of smooth muscle relaxation.
- Computational docking supported experimental findings by identifying ligand-binding sites on major ion channels.
- The integrated experimental–computational approach provides insights for developing targeted cardiovascular therapeutics.
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PY Chen, L Qin, G Li, J Malagon-Lopez, Z Wang, S Bergaya, S Gujja, AW Caulk, SI Murtada, X Zhang, ZW Zhuang, DA Rao, G Wang, Z Tobiasova, B Jiang, RR Montgomery, L Sun, H Sun, EA Fisher, JR Gulcher, C Fernandez-Hernando, JD Humphrey, G Tellides, TW Chittenden and M Simons. Smooth muscle cell reprogramming in aortic aneurysms. Cell Stem Cell: Cell Press 20202; 26(4), 542-557.
J Redon and R Carmena. Present and future of drug therapy in hypertension: An overview. Blood Pressure 2024. https://doi.org/10.1080/08037051.2024.2320401
C Chan and DHH Lau. Challenges in the clinical care of major cardiovascular conditions in primary care: A narrative review. Current Problems in Cardiology 2025; 50(8), 103099.
Z Wang, X Sun, M Sun, C Wang and L Yang. Game changers: Blockbuster small-molecule drugs approved by the FDA in 2024. Pharmaceuticals 2025; 18(5), 729.
L Muhl, G Mocci, R Pietilä, J Liu, L He, G Genové, S Leptidis, S Gustafsson, B Buyandelger, E Raschperger, EM Hansson, JLM Björkegren, M Vanlandewijck, U Lendahl and C Betsholtz. A single-cell transcriptomic inventory of murine smooth muscle cells. Developmental Cell 2022; 57(20), 2426-2443.e6.
S Sodiqova, S Kadirova, A Zaynabiddinov, I Abdullaev, L Makhmudov, U Gayibov, M Yuldasheva, M Xolmirzayeva, R Rakhimov, A Mutalibov and H Karimjonov. Channelopathy activity of A-41(Propyl Ester of Gallic Acid): Experimental and computational study of antihypertensive activity. Trends in Sciences 2025; 22(9), 10496.
TF Aripov and UG Gayibov. Antiradical and antioxidant activity of the preparation “Rutan” from Rhus coriaria L. Journal of Theoretical and Clinical Medicine 2023;4, 164-170.
B Kim, C Jo, HY Choi and K Lee. Prunetin relaxed isolated rat aortic rings by blocking calcium channels. Molecules 2018; 23(9), 2372.
B Kim, K Lee, KS Chinannai, I Ham, Y Bu, H Kim, HY Choi. Endothelium-independent vasorelaxant effect of Ligusticum jeholense root and rhizoma on rat thoracic aorta. Molecule 2015; 20(6), 10721-10733.
ML Addison, P Ranasinghe and DJ Webb. Novel pharmacological approaches in the treatment of hypertension: A focus on RNA-based therapeutics. Hypertension 2023; 80(11), 2243-2254.
R Sayidaliyeva, S Kadirova, A Zaynabiddinov, I Abdullaev, L Makhmudov, U Gayibov, M Yuldasheva, M Kholmirzayeva, R Rakhimov, A Mutalibov and H Karimjonov. A-51 as a natural calcium channel blocker: An integrative study targeting hypertension. Trends in Sciences 2025; 22(11), 10760.
E Altobelli; L Rapacchietta, VF Profeta and R Fagnano. Risk factors for abdominal aortic aneurysm in population-based studies: A systematic review and meta-analysis. International Journal of Environmental Research and Public Health 2018; 15(12), 2805.
M Zaripova, I Abdullaev, A Bogbekov, U Gayibov, S Omonturdiev, R Makhmudov, N Ergashev, G Jabbarova, S Gayibova and T Aripov. In Vitro and in Silico studies of Gnaphalium U. Extract: Inhibition of α-amylase and α-glucosidase as a potential strategy for metabolic syndrome regulation. Trends in Sciences 2025; 22(8), 10098.
FV Brozovich, CJ Nicholson, CV Degen, YZ Gao, M Aggarwal and KG Morgan. Mechanisms of vascular smooth muscle contraction and the basis for pharmacologic treatment of smooth muscle disorders. Pharmacological Reviews 2016; 68(2), 476-532.
OS Zoirovich, AIZ Ugli, ID Raxmatillayevich, ML Umarjonovich, ZM Ravshanovna, GS Narimanovna, GU Gapparjanovich and AT Fatikhovich. The effect of Ájuga Turkestánica on the rat aortic smooth muscle ion channels. Biomedical & Pharmacology Journal 2024; 17(2), 1213-1222.
G Cao, X Xuan, J Hu, R Zhang, H Jin and H Dong. How vascular smooth muscle cell phenotype switching contributes to vascular disease. Cell Communication and Signaling 2022; 20(1), 180.
D Ghosh, AU Syed, MP Prada, MA Nystoriak, LF Santana, M Nieves-Cintrón and MF Navedo. Calcium channels in vascular smooth muscle. Advances in Pharmacology 2017; 78, 49-87.
YL Chen, Z Daneva, M Kuppusamy, M Ottolini, TM Baker, E Klimentova, SA Shah, JD Sokolowski, MS Park and SK Sonkusare. Novel smooth muscle Ca(2+)-signaling nanodomains in blood pressure regulation. American Heart Association 2022; 146(7), 548-564.
I McFadzean and A Gibson. The developing relationship between receptor-operated and store-operated calcium channels in smooth muscle. British Journal of Pharmacology 2002; 135(1), 1-13.
Q Lin, G Zhao, X Fang, X Peng, H Tang, H Wang, R Jing, J Liu, WJ Lederer, J Chen and K Ouyang. IP3 receptors regulate vascular smooth muscle contractility and hypertension. JCI Insight 2016; 1(17), e89402.
Z Liu, RA Khalil. Evolving mechanisms of vascular smooth muscle contraction highlight key targets in vascular disease. Biochemical Pharmacology 2018; 153, 91-122.
WF Jackson. KV channels and the regulation of vascular smooth muscle tone. Microcirculation 2018; 25(1), e12421.
D Guntur, H Olschewski, P Enyedi, R Csáki, A Olschewski and C Nagaraj. Revisiting the large-conductance calcium-activated potassium (BKCa) channels in the pulmonary circulation. Biomolecules 2021; 11(11), 1629.
KP Nunes and RC Webb. New insights into RhoA/Rho-kinase signaling: A key regulator of vascular contraction. Small GTPases 2021; 12(5-6), 458-469.
M Ottolini and SK Sonkusare. The calcium signaling mechanisms in arterial smooth muscle and endothelial cells. Comprehensive Physiology 2021; 11(2), 1831-1869.
AV Mahmudov, OS Abduraimov, SB Erdonov, UG Gayibov and LY Izotova. Bioecological features of nigella sativa L. in different conditions of Uzbekistan. Plant Science Today 2022; 9(2): 421-426.
U Gayibov, I Abdullaev, F Sobirova, S Omonturdiev, A Abdullaev, S Gayibova and T Aripov. Plant-derived and synthetic antihypoxic agents in cardiovascular diseases: Mechanisms, key pathways and therapeutic potential. Plant Science Today 2025; 12(4), 1-10.
D Inomjonov, I Abdullaev, S Omonturdiev, A Abdullaev, L Maxmudov, M Zaripova, M Abdullayeva, D Abduazimova, S Menglieva, S Gayibova, Ma Sadbarxon, U Gayibov and T Aripov. In Vitro and In Vivo studies of crategus and inula helenium extracts: Their effects on rat blood pressure. Trends in Sciences 2025, 22(3), 9158
S D’Haese, D Deluyker and V Bito. Acute exposure to glycated proteins impaired in the endothelium-dependent aortic relaxation: A matter of oxidative stress. International Journal of Molecular Sciences 2022; 23(23), 14916.
YC Loh, CS Tan, YS Ch’ng, M Ahmad, MZ Asmawi and MF Yam. Overview of antagonists used for determining the mechanisms of action employed by potential vasodilators with their suggested signaling pathways. Molecules. 2016; 21(4), 495.
I Abdullaev, U Gayibov, S Omonturdiev, S Fotima, S Gayibova and T Aripov. Molecular pathways in cardiovascular disease under hypoxia: Mechanisms, biomarkers, and therapeutic targets[J]. The Journal of Biomedical Research 2025; 39(3): 254-269.
A Arsyad, GKR Lembang, SL Linda, YY Djabir and GP Dobson. Low calcium-high magnesium krebs-henseleit solution combined with adenosine and lidocaine improved rat aortic function and structure following cold preservation. Medicina (Kaunas) 2024; 60(8), 1284.
CJ Hansen, S Siricilla, N Boatwright, JH Rogers, ME Kumi and J Herington. Effects of solvents, emulsions, cosolvents, and complexions on ex vivo mouse myometrial contractility. Reproductive Sciences 2022; 29(2), 586-595.
MAS Leal, TD Almeida, JG Torres, LCG Campos, EC Vasquez and VG Barauna. Physiological and biochemical vascular. reactivity parameters of angiotensin ii and the action of biased agonist TRV023. Advances in Pharmacological and Pharmaceutical Sciences 2020; 2020, 3092721.
A Esimbetov, A Zaripov, S Omonturdiev, M Sultankhodjaev and P Usmanov. vasorelaxant effect of the alkaloid 1-0-Benzoylnappeline on isolated rat thoracic aorta. International Journal of Agriculture, Environment and Bioresearch 2020; 5(2), 146-156.
A Abdullaev, I Abdullaev, A Bogbekov, U Gayibov, S Omonturdiev, S Gayibova, M Turahodjayev, K Ruziboev and T Aripov. Antioxidant potential of rhodiola heterodonta extract: Activation of Nrf2 pathway via integrative In Vivo and In Silico studies. Trends in Sciences, 2025; 22(5), 9521.
MR Zaripova, SN Gayibova, RR Makhmudov, AA Mamadrahimov, Nl Vypova, UG Gayibov, SM Miralimova and TF Aripov. Characterization of Rhodiola heterodonta (Crassulaceae): Phytocomposition, antioxidant and antihyperglycemic activities. Preventive Nutrition and Food Science 2024; 29(2), 135-145.
K Karabacak, E Kaya, KG Ulusoy, M Seyrek, M Kurtoglu, S Doganci, V Yildirim, O Yildiz and U Demirkilic. Effects of taurine on contractions of human internal mammary artery: A potassium channel opening action. European Review for Medical and Pharmacological Sciences 2015; 19, 1498-1504.
AV Mahmudov, OS Abduraimov, SB Erdonov, AL Allamurotov, OT Mamatqosimov, UG Gayibov and L Izotova. Seed productivity of Linum usitatissimum L. in different ecological conditions of Uzbekistan. Plant Science Today 2022; 9(4), 1090-1101.
T Godfraind. Discovery and development of calcium channel blockers. Frontiers in pharmacology 2017; 8, 286.
U Gayibov, SN Gayibova, MK Pozilov, FS Tuxtaeva, UR Yusupova, GMK DJabbarova, ZA Mamatova, NA Ergashev and TF Aripov. Influence of quercetin and dihydroquercetin on some functional parameters of rat liver mitochondria. Journal of Microbiology, Biotechnology and Food Sciences 2021; 11(1), e2924.
L Collins, L Lam, O Kleinig, W Proudman, R Zhang, M Bagster, J Kovoor, A Gupta, R Goh, S Bacchi, D Schultz and T Kleinig. Verapamil in the treatment of reversible cerebral vasoconstriction syndrome: A systematic review. Journal of Clinical Neuroscience: Official Journal of the Neurosurgical Society of Australasia 2023; 113, 130-141.
UG Gayibov. Influence of new polyphenol compound from Euphorbia plant on mitochondrial function. Journal of Microbiology, Biotechnology and Food Sciences 2019; 8(4), 1021-1025.
S Koli, A Prakash, S Choudhury, R Mandil and SK Garg. Calcium Channels, Rho-Kinase, Protein Kinase-C, and Phospholipase-C Pathways Mediate Mercury Chloride-Induced Myometrial Contractions in Rats. Biological Trace Element Research 2019; 187(2), 418-424.
MK Pozilov, U Gayibov, MI Asrarov, NG Abdulladjanova, HS Ruziboev and TF Aripov. Physiological alterations of mitochondria under diabetes condition and its correction by polyphenol gossitan. Journal of Microbiology, Biotechnology and Food Sciences 2022; 12(2), e2224.
AG Vakhobjonovna, KE Jurayevich, AIZ Ogli, EN Azamovich, MR Rasuljonovich and AM Islomovich. Tannins as modulators in the prevention of mitochondrial dysfunction. Trends in Sciences 2025; 22(8), 10436.
I Abdurazakova, A Zaynabiddinov, I bdullaev, L Makhmudov, U Gayibov, S Omonturdiev, G Abdullayev, M Xolmirzayeva and S Zhurakulov. Pharmacological evaluation of F-45 on the cardiovascular system using in vitro, in vivo models and molecular dockings. Trends in Sciences 2025; 22(12), 10924.
CA D’Addario, S Matsumura, A Kitagawa, GM Lainer, F Zhang, M D’silva, MY Khan, G Froogh, A Gruzdev, DC Zeldin, ML Schwartzman and SA Gupte. Global and endothelial G-protein coupled receptor 75 (GPR75) knockout relaxes pulmonary artery and mitigates hypoxia-induced pulmonary hypertension. Vascular Pharmacology 2023; 153, 107235.
M Mamajanov, I Abdullaev, G Sotimov, S Mavlanova, Q Niyozov, M Mirzaolimov, A Najimov, E Mirzaolimov, M Raximberganov and U Abdullayev. Mitochondrial and pharmacokinetic insights into 3,5,7,2′,6′-pentahydroxyflavanone: respiratory modulation, calcium handling, and membrane stability. Trends in Sciences 2025; 22(12), 10984.
EV Hersh, PA Moore and M Saraghi. Phentolamine mesylate: Pharmacology, efficacy, and safety. General Dentistry 2019; 67(3), 12-17.
Z Shakiryanova, R Khegay, U Gayibov, A Saparbekova, Z Konarbayeva, A Latif and O Smirnova. Isolation and study of a bioactive extract enriched with anthocyanin from red grape pomace (Cabernet sauvignon). Agronomy Research 2023; 21(3), 1293-1303.
R Zhang, L Peng, H Ran, Y Fan, Y Zhao, F Cao. Farnesoid X receptor activation modulates calcium homeostasis in rat aortic vascular smooth muscle cells. The Chinese Journal of Physiology 2018; 61(4), 210-220.
K Rahate, LK Bhatt and KS Prabhavalkar. SERCA stimulation: A potential approach in therapeutics. Chemical Biology & Drug Design 2020; 95(1), 5-15.
R Dahl and I Bezprozvanny. SERCA pump as a novel therapeutic target for treating neurodegenerative disorders. Biochemical and Biophysical Research Communications 2024; 734,150748.
PJ Chambers, ES Juracic, VA Fajardo and AR Tupling. Role of SERCA and sarcolipin in adaptive muscle remodeling. American Journal of Physiology, Cell physiology 2022; 322(3), C382-C394.
C Hidalgo and A Paula-Lima. RyR-mediated calcium release in hippocampal health and disease. Trends in Molecular Medicine 2024; 30(1), 25-36.
Gayibov UG, Gayibova SN, Karimjonov HM, Abdullaev AA, et al. Antioxidant and cardioprotective properties of polyphenolic plant extract of Rhus glabra L. Plant Sci Today. 2024;11(3), 2348–1900.
DR Laver. Regulation of the RyR channel gating by Ca2+ and Mg2. Biophysical Reviews 2018; 10(4), 1087-1095.
S Gleitze, OA Ramírez, I Vega-Vásquez,J Yan, P Lobos, H Bading, MT Núñez, A Pauka-Lima and C Hidalgo. Ryanodine receptor mediated calcium release contributes to ferroptosis induced in primary hippocampal neurons by GPX4 inhibition. Antioxidants (Basel) 2023; 12(3), 705.
AGS Harper and SO Sage. TRP-Na(+)/Ca(2+) exchanger coupling. Advances in Experimental Medicine and Biology 2016; 898, 67-85.
T Loeck and A Schwab. The role of the Na+/Ca2+-exchanger (NCX) in cancer-associated fibroblasts. Biological Chemistry 2023; 404(4), 325-337.
RG Contreras, A Torres-Carrillo, C Flores-Maldonado, L Shoshani and A Ponce. Na+/K+-ATPase: More than an electrogenic pump. International Journal of Molecular Sciences 2024; 25(11), 6122.
O Gaibullayeva, A Islomov, D Abdugafurova, B Elmurodov, B Mirsalixov, L Mahmudov, I Adullaev, K Baratov, S Omonturdiev and S Sa’dullayeva. Inula helenium L. root extract in sunflower oil: Determination of its content of water-soluble vitamins and immunity-promoting effect. Biomedical Pharmacology Journal 2024; 17(4), 2729-2737.
Y Umidakhon, B Erkin, G Ulugbek, et al. Correction of the mitochondrial NADH oxidase activity, peroxidation and phospholipid metabolism by haplogenin-7-glucoside in hypoxia and ischemia[J]. Trends Sci 2022, 19(21), 6260.
RJ Korthuis. Mechanisms of I/R-Induced endothelium-dependent vasodilator dysfunction. Advances in Pharmacology 2018; 81, 331-364.
AA Abdullaev, DR Inamjanov, DS Abduazimova, SZ Omonturdiyev, UG Gayibov, SN Gayibova and TF Aripov. Sílybum Mariánum’s impact on physiological alterations and oxidative stress in diabetic rats. Biomedical and Pharmacology Journal 2024; 17(2), 1291-1300.
M Li, M Zhang, AT Fatixovich, GU Gapparjanovich and H Du. Green-synthesized Zn²⁺-polyphenol networks (CGA/RA) for enhanced multifunctional food preservation. Journal of Food Measurement and Characterization 2025. https://doi.org/10.1007/s11694-025-03764-y
UG Gayibov, SN Gayibova , HM Karimjonov, AA Abdullaev, DS Abduazimova, RN Rakhimov, HS Ruziboev, MA Xolmirzayeva, AE Zaynabiddinov and TF Aripov. Antiradical and antioxidant activity of the preparation “Rutan” from Rhus coriaria L. Journal of Theoretical and Clinical Medicine 2023; 4, 164-170.
LHS Santos, RS Ferreira and ER Caffarena. Integrating molecular docking and molecular dynamics simulations. Methods in Molecular Biology 2019; 2053, 13-34.
P Peluso, A Dessì, R Dallocchio, V Mamane and S Cossu. Recent studies of docking and molecular dynamics simulation for liquid-phase enantioseparations. Electrophoresis 2019; 40(15), 1881-1896.
AQQ Azimova, AX Islomov, SA Maulyanov, D Gulyamovna Abdugafurova, LU Mahmudov, IZiyoyiddin Abdullaev, AS Ishmuratova, SQ Siddikova and IR Askarov. Determination of vitamins and pharmacological properties of Vitis vinifera L. plant fruit part (mixed varieties) syrup-honey. Biomedical and Pharmacology Journal 2024; 17(4), 2779-2786.
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