scholarly journals Gentiopicroside Produces Endothelium-Independent Vasodilation by Deactivating the PI3K/Akt/Rho-Kinase Pathway in Isolated Rat Thoracic Aorta

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Shangping Xing ◽  
Feifei Nong ◽  
Jialiang Qin ◽  
Huicai Huang ◽  
Ruoting Zhan ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Kinase Inhibitor ◽  
Rho Kinase ◽  
Ruthenium Red ◽  
Akt Inhibitor ◽  
Vasorelaxant Effect ◽  
Protein Levels ◽  
Cox Inhibitor ◽  
Rat Thoracic Aorta ◽  
Isolated Rat

Gentiopicroside (GPS), a main active secoiridoid glucoside derived from the roots of perennial herbs in the Gentianaceae family, has antispasmodic and relaxant effects. However, the vasorelaxant effects of GPS on aortic rings and the molecular mechanisms involved in these effects are not yet clear. Therefore, we investigated whether GPS inhibits phenylephrine- (PE-) or KCl-induced contractions in isolated rat thoracic aortic rings. The present study found that GPS produced a dose-dependent relaxation in aortic rings precontracted with PE or KCl and significantly reduced CaCl2-, narciclasine- (Rho-kinase activator-), and phorbol-12,13-diacetate- (PKC activator-) induced vasocontractions. Pretreatment with NG-Nitroarginine methyl ester hydrochloride (L-NAME, NOS inhibitor), methylene blue (sGC inhibitor), indomethacin (COX inhibitor), 4-aminopyridine (KV channel inhibitor), and glibenclamide (KATP channel inhibitor) had no influence on the vasorelaxant effect of GPS, while BaCl2 (Kir channel inhibitor), tetraethylammonium chloride (KCa channel inhibitor), ruthenium red (RYR inhibitor), and heparin (IP3R inhibitor) significantly reduced GPS-induced vasorelaxation. Moreover, GPS pretreatment remarkably inhibited the influx of Ca2+ in vascular smooth muscle cells stimulated using KCl or PE-containing CaCl2 solution. Western blot analysis confirmed that GPS treatment inhibited PE-induced increases in the protein levels of p-Akt, p-myosin light chain (MLC), and p-myosin-binding subunit of myosin phosphatase 1 (MYPT1) in the aortic rings. Additionally, the vasorelaxation activity of GPS was attenuated upon pretreatment with LY294002 (PI3K/Akt inhibitor), Y27632 (Rho-kinase inhibitor), and verapamil (L-type Ca2+ channel inhibitor). These findings demonstrate that GPS exhibits endothelium-independent vasorelaxant effects through inhibition of voltage-dependent, receptor-operated, and inositol triphosphate receptor (IP3R)/ryanodine receptor- (RYR-) mediated Ca2+ channels as well as the PI3K/Akt/Rho-kinase signaling pathway.

scholarly journals Bubbles Induce Endothelial Microparticle Formation via a Calcium-Dependent Pathway Involving Flippase Inactivation and Rho Kinase Activation

2018 ◽  
Vol 46 (3) ◽  
pp. 965-974 ◽  
Author(s):  
Xuhua Yu ◽  
Jiajun Xu ◽  
Wenwu Liu ◽  
Weigang Xu
Keyword(s):  
Kinase Inhibitor ◽  
Umbilical Vein ◽  
Vascular Inflammation ◽  
Cell Communication ◽  
Rho Kinase ◽  
Cytoplasmic Calcium ◽  
Kinase Substrate ◽  
Protein Levels ◽  
Underlying Mechanisms ◽  
Umbilical Vein Endothelial Cells

Background/Aims: Intravascular bubbles can exert pleiotropic detrimental effects, partly by inducing endothelial microparticles (EMPs) production, which play critical roles in cell communication and vascular inflammation cascades. However, the underlying mechanisms remain unclear. This study aimed to delineate the possible mechanisms involving bubble-induced EMPs formation. Methods: Human umbilical vein endothelial cells (HUVECs) were contacted by bubbles and EMPs level in supernatant were quantified by flow cytometry. Cytoplasmic calcium (Ca2+) was measured by the Ca2+ binding dyes Fluo-3 AM and flippase activity was assessed by translocation rate of fluorescent phosphatidylserine (PS) analogue NBD-PS. Protein levels of phospho-myosin light chain (MLC, a Rho kinase substrate) and phospho-extracellular signal-regulated kinase 1 or 2 (ERK1/2) were determined by western blotting. The score of actin colocalization was assessed by phalloidin-FITC using an immunofluorescent microscopy. Results: EMPs level markedly increased after bubble stimulus. Cytoplasmic calcium (Ca2+) significantly elevated (P< 0.05), flippase activity decreased (P< 0.05), protein levels of phospho- MLC and phospho- ERK1/2 significantly increased (P< 0.05, P < 0.05), and the score of actin colocalization markedly reduced (P< 0.05) in bubble-stimulated HUVECs. All the above changes except the increase in phospho-ERK1/2 can be reversed by Ca2+ channel blocker LaCl3 (P< 0.05). Additionally, MLC phosphorylation was significantly inhibited and actin colocalization markedly increased by Rho kinase inhibitor pretreatment and more importantly, bubble-induced EMPs markedly decreased. Conclusions: These results demonstrate that bubble stimulates EMPs formation by cytoplasmic Ca2+ elevation and subsequently activating Rho kinase pathway and cytoskeleton reorganization. Simultaneously, cytoplasmic Ca2+ inhibits the flippase activity and subsequently increases phosphatidylserine exposure, which also contributes to EMPs formation.

Effect of fasudil on growth, adhesion, invasion, and migration of 95D lung carcinoma cells in vitro

2010 ◽  
Vol 88 (9) ◽  
pp. 874-879 ◽  
Author(s):  
Xueying Yang ◽  
Yang Zhang ◽  
Shumin Wang ◽  
Wenjun Shi
Keyword(s):  
Lung Carcinoma ◽  
Molecular Mechanisms ◽  
Kinase Inhibitor ◽  
Rho Kinase ◽  
Carcinoma Cells ◽  
Migration And Invasion ◽  
Invasion And Migration ◽  
Lung Carcinoma Cells ◽  
Rho Kinase Inhibitor ◽  
And Migration

The objective of the study was to investigate the effects of a Rho-kinase inhibitor on 95D lung carcinoma cell growth, adhesion, invasion, and migration and to explore the underlying molecular mechanisms involved in this process. After treatment of 95D lung carcinoma cells with fasudil, an inhibitor of Rho-kinase, cell biological behaviors such as growth, adhesion, invasion, and migration were observed. Matrix metalloproteinase (MMP) activity and Western blot assay were used to evaluate underlying molecular mechanisms. The IC50 of fasudil to 95D lung carcinoma cells was approximately 0.79 mg/mL (95% confidence limits 0.58–1.11 mg/mL). After treatment with 0.75 mg/mL fasudil, the ability of 95D lung carcinoma cells for growth, adhesion, migration, and invasion was decreased significantly. Total active MMP2 was decreased approximately 22.7% (p < 0.05) and total MMP9 65.9% (p < 0.01). Myosin phosphatase target subunit 1 (MYPT1) was reduced by 29.4% (p < 0.05). We conclude that the Rho-kinase inhibitor prevents the growth, adhesion, invasion, and migration of 95D lung carcinoma cells by inhibiting the Rho/Rho-kinase pathway. Changes in MMP2, MMP9, and MYPT1 may be part of its molecular mechanisms.

Improved erectile function after Rho-kinase inhibition in a rat castrate model of erectile dysfunction

2003 ◽  
Vol 284 (6) ◽  
pp. R1572-R1579 ◽  
Author(s):  
Christopher J. Wingard ◽  
John A. Johnson ◽  
Andre Holmes ◽  
Anita Prikosh
Keyword(s):  
Nitric Oxide ◽  
Erectile Dysfunction ◽  
Erectile Function ◽  
Kinase Inhibitor ◽  
Rho Kinase ◽  
Voltage Response ◽  
Intracavernosal Injection ◽  
Protein Levels ◽  
Erectile Response ◽  
Kinase Pathway

Androgens are reported to act as strong modulators of erectile function influencing both nitric oxide and vasoconstrictor signaling. Castration results in a depressed erectile response that is associated with a loss of nitric oxide production and increased responsiveness to constrictive agents. The increased vasoconstrictor response may be a result of an active RhoA/Rho-kinase signaling pathway. We report here results of studies designed to test the hypothesis that inhibition of the Rho-kinase pathway restores erectile function in a castrate model by relaxing the smooth muscle. Mean arterial (MAP) and corpus cavernosal (CCP) pressures were monitored during intracavernosal injection of the Rho-kinase inhibitor Y-27632. Castration reduced the maximal erectile response (CCP/MAP) by 33%, and testosterone replacement restored the response (intact, 0.736 ± 0.040; castrate, 0.492 ± 0.022; testosterone, 0.681 ± 0.073). Injection of Y-27632 increased CCP in all experimental groups; it also left shifted the voltage response curve and increased the maximal CCP/MAP response (intact, 0.753 ± 0.091; castrate, 0.782 ± 0.081; testosterone treated, 0.894 ± 0.033). Y-27632 dose dependently relaxed phenylephrine-stimulated cavernosal tissues. Cavernosal tissues showed increased RhoA and Rho-kinase protein levels after castration. Our data support the hypothesis that an active Rho/Rho-kinase pathway contributes to the reduced erectile response after castration due to an upregulation of RhoA/Rho-kinase protein levels and that inhibition of this pathway may serve as an effective treatment for erectile dysfunction.

scholarly journals VASORELAXANT EFFECT OF THE ALKALOID 1-0-BENZOYLNAPPELINE ON ISOLATED RAT THORACIC AORTA

2020 ◽  
Vol 05 (02) ◽  
pp. 146-156
Author(s):  
Adilbay Esimbetov ◽  
Abdisalim Zaripov ◽  
Sirojiddin Omonturdiev ◽  
Mukhlis Sultankhodjaev ◽  
Pulat Usmanov
Keyword(s):  
Thoracic Aorta ◽  
Vasorelaxant Effect ◽  
Rat Thoracic Aorta ◽  
Isolated Rat

Contributions of Rho-kinase and AMP-related kinase signaling pathways to responses mediated by endothelium-derived contracting factors in diabetic rat aorta

2019 ◽  
Vol 97 (7) ◽  
pp. 600-610 ◽  
Author(s):  
Cennet Balcilar ◽  
Isil Ozakca-Gunduz ◽  
V. Melih Altan
Keyword(s):  
Signaling Pathways ◽  
Vascular Function ◽  
Kinase Inhibitor ◽  
Rho Kinase ◽  
Calcium Ionophore ◽  
Diabetic Rats ◽  
Diabetic Rat ◽  
Relaxation Response ◽  
Ionophore A23187 ◽  
Protein Levels

Diabetes-induced endothelial damage leads to vascular dysfunction. The current study investigated the effects of short-term (4-week) streptozotocin (STZ)-induced diabetes on responses mediated by endothelium-derived contracting factors (EDCFs) as well as possible contributions of Rho-kinase and AMP-activated kinase (AMPK) signaling pathways. The effects of STZ-diabetes on vascular function were examined in isolated thoracic aorta preparations of 30-week-old rats (n = 27). The diabetes-associated changes in vascular function were studied with calcium ionophore A23187, acetylcholine, Rho-kinase inhibitor Y27632 ((R)-(+)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride), and AMPK activator AICAR (5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside). The phosphorylation of acetyl-CoA carboxylase, AMPK, and phospholamban and the protein levels of sarcoplasmic/endoplasmic Ca2+-ATPase 2 (SERCA2) and Rho-associated protein kinase (ROCKII) were measured in aortic preparations. Although the acetylcholine-mediated relaxation responses were preserved in 4-week STZ-induced diabetes, the increased activation of the Rho-kinase pathway was demonstrated via twofold enhancement in A23187-mediated contractile responses and significantly augmented protein levels of ROCKII. The AICAR-activated AMPK-mediated relaxation response was also augmented ∼4-fold in diabetic rats, without any alteration in phospholamban phosphorylation; further, this relaxation response suppressed A23187-mediated contraction in both groups. Diabetic rats showed an increase in AICAR-induced AMPK-mediated vasorelaxation and a 2.5-fold elevation of phosphorylated AMPK levels. These results indicate a possible compensation between hyperglycemia-induced endothelium-dependent hypercontractility and AMPK-mediated vasorelaxation in diabetes.

Vasorelaxant effect of the flavonoid galangin on isolated rat thoracic aorta

Life Sciences ◽  
2006 ◽  
Vol 78 (8) ◽  
pp. 825-830 ◽  
Author(s):  
Silvana Morello ◽  
Valentina Vellecco ◽  
Alessio Alfieri ◽  
Nicola Mascolo ◽  
Carla Cicala
Keyword(s):  
Thoracic Aorta ◽  
Vasorelaxant Effect ◽  
Rat Thoracic Aorta ◽  
Isolated Rat

Endothelin-1-induced contraction in veins is independent of hydrogen peroxide

2005 ◽  
Vol 289 (3) ◽  
pp. H1115-H1122 ◽  
Author(s):  
Keshari Thakali ◽  
Stacie L. Demel ◽  
Gregory D. Fink ◽  
Stephanie W. Watts
Keyword(s):  
Vascular Tone ◽  
Kinase Inhibitor ◽  
Vena Cava ◽  
Rho Kinase ◽  
Oxygen Species ◽  
Endothelin 1 ◽  
Rho Kinase Inhibitor ◽  
Rat Thoracic Aorta ◽  
Vascular Beds ◽  
Kinase Pathway

Reactive oxygen species (ROS), such as superoxide and H2O2, are capable of modifying vascular tone, although the response to ROS can vary qualitatively among vascular beds, experimental procedures, and species. Endothelin-1 (ET-1) induces superoxide production, which can be dismutated to H2O2. The RhoA/Rho kinase pathway partially mediates ET-1-induced contraction and recently was implicated in superoxide-induced contraction. We hypothesized that H2O2, not superoxide, mediates venous ET-1-induced contraction. Rat thoracic aorta and vena cava contracted to exogenously added H2O2 (1 μM–1 mM) [maximum aortic contraction = 10 ± 3% of phenylephrine (10 μM) contraction; maximum venous contraction = 85 ± 13% of norepinephrine (10 μM) contraction]. (+)-( R)- trans-4-(1-aminoethyl- N-4-pyridil)cyclohexanecarboxamide dihydrochloride (Y-27632, 10 μM), a Rho kinase inhibitor, significantly reduced venous H2O2-induced contraction (15 ± 1% of control maximum) and reduced maximum ET-1-induced contraction by 59 ± 1%. However, neither the H2O2 scavenger catalase (100 and 2,000 U/ml) nor cell permeable polyethylene glycol-catalase (163 and 326 U/ml) reduced ET-1-induced contraction in the vena cava. The catalase inhibitor 3-aminotriazole (3-AT) also had no effect on maximal venous ET-1-induced contraction. Basal H2O2 levels were three times higher in the vena cava than in the aorta (vena cava, 0.74 ± 0.09 nmol H2O2/mg protein; aorta, 0.24 ± 0.05 nmol H2O2/mg protein). ET-1 (100 nM) increased H2O2 in the vena cava but not in the aorta (vena cava, 154.10 ± 17.29% of control H2O2; aorta, 83.72 ± 20.20%). Antagonism of either ETA or ETB receptors with the use of atrasentan (30 nM) or BQ-788 (100 nM), respectively, reduced ET-1 (100 nM)-induced increases in venous H2O2. In summary, ET-1 increased H2O2 in veins but not arteries, and venous ET-1-induced H2O2 production was independent of the contractile properties of ET-1.

Protective Effect of Nebulized Rho-Kinase Inhibitor on Ischemia Reperfusion Injury in Isolated Rat Lung Perfusion Model

2015 ◽  
Vol 34 (4) ◽  
pp. S270-S271
Author(s):  
K. Ohata ◽  
F. Chen ◽  
M. Takahashi ◽  
T. Kondo ◽  
H. Motoyama ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Rho Kinase ◽  
Lung Perfusion ◽  
Rat Lung ◽  
Perfusion Model ◽  
Rho Kinase Inhibitor ◽  
Isolated Rat Lung ◽  
Isolated Rat

Vasorelaxant Effect of the Rootbark Extract of Paeonia moutan on Isolated Rat Thoracic Aorta

Planta Medica ◽  
2006 ◽  
Vol 72 (14) ◽  
pp. 1338-1341 ◽  
Author(s):  
Mi Yoo ◽  
Byung Lee ◽  
Yeon Choi ◽  
Jung Lee ◽  
Jee Seo ◽  
...  
Keyword(s):  
Thoracic Aorta ◽  
Vasorelaxant Effect ◽  
Rat Thoracic Aorta ◽  
Isolated Rat

scholarly journals NLRP3 maintains healthy pericytes in the brain

2019 ◽  
Author(s):  
Wenqiang Quan ◽  
Qinghua Luo ◽  
Qiqiang Tang ◽  
Tomomi Furihata ◽  
Dong Li ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Collagen Type Iv ◽  
Akt Inhibitor ◽  
Wild Type Littermate ◽  
Cerebral Microvessels ◽  
Protein Levels ◽  
Type Iv ◽  
Brain Pericytes ◽  
And Function ◽  
The Brain

Abstract Background Pericytes regulate structure and function of cerebral capillaries. Growing evidence shows that pericytes are damaged in the brain of Alzheimer’s disease (AD), which potentially contributes to AD pathogenesis. NLRP3-contained inflammasome is activated in AD brain and considered as a promising target for therapy. However, how NLRP3 affects brain pericytes is unclear. In our study, we investigated physiological function of NLRP3 in pericytes. Methods Immunohistological methods and Western blot were used to investigate pericytes and vasculature in the brains of 9-month-old NLRP3-deficient and wild-type littermate mice. Pericytes were also cultured and treated with NLRP3 inhibitor, recombinant IL-1β and AKT inhibitor. Then, proliferation, apoptosis and expression of PDGFRβ and CD13 in pericytes were analysed with biochemical methods. To investigate underlying molecular mechanisms, phosphorylation of protein kinases such as AKT, ERK and NF- k B were quantified. Results We observed that NLRP3 deficiency reduced the coverage of PDGFRβ-positive pericytes and collagen type IV-immunereactive vasculature in the brain. NLRP3 deficiency was also shown to decrease PDGFRβ and CD13 proteins in isolated cerebral microvessels. In cultured pericytes, inhibition of NLRP3 with MCC950 attenuated cell proliferation but did not induce apoptosis. NLRP3 inhibition also decreased protein levels of PDGFRβ and CD13. On the contrary, treatments with IL-1β increased protein levels of PDGFRβ and CD13 in pericytes. The alteration of PDGFRβ and CD13 protein levels was correlated with phosphorylation of AKT. Inhibition of AKT reduced PDGFRβ and CD13 in cultured pericytes. Conclusions NLRP3 might be essential to maintain healthy pericytes in the brain through activating AKT. Adverse effects on brain pericytes should be considered in the possible clinical therapies with NLRP3 inhibitors.

Sign in / Sign up

Export Citation Format

Share Document