Rho kinase activation in circulating leukocytes is related to hypertensive myocardial remodeling

2018 ◽  
Vol 132 (16) ◽  
pp. 1837-1853 ◽  
Author(s):  
Maria P. Ocaranza ◽  
Camila Fierro ◽  
Jorge E. Jalil ◽  
Jackeline Moya ◽  
Leticia Gonzalez ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Sham Group ◽  
Kinase Activity ◽  
Aortic Wall ◽  
Rho Kinase ◽  
Myocardial Remodeling ◽  
Pathway Activation ◽  
Kinase Cascade ◽  
Degree Of Association ◽  
Myocardial Collagen

Rho-kinase has relevant functions in blood pressure modulation and cardiovascular remodeling. Rho-kinase activity is determined in circulating leukocytes measuring phosphorylation of its target myosin phosphatase target subunit 1 (MYPT1), but its relationship with Rho-kinase activity in the myocardium and in vasculature in hypertension has not been evaluated. The aim was to determine the degree of association between Rho-kinase cascade activation in circulating leukocytes with cardiac and aortic Rho-kinase pathway activation in a model of hypertension and to analyze it with a cause–effect perspective. Hypertensive deoxycorticosterone (DOCA)-salt rats received the Rho-kinase antagonist fasudil (DOCA-Fas, 100 mg/kg/day, 3 weeks). Results were compared with an untreated DOCA-salt and a sham group. Rho-kinase inhibition reduced significantly blood pressure, cardiac hypertrophy, myocardial collagen and macrophage infiltration, but not aortic wall hypertrophy. Fasudil decreased significantly Rho-kinase activity in peripheral blood mononucleated cells (PBMC), myocardium and aortic wall to similar levels as in the sham group. A significant correlation was found between PBMC Rho-kinase activity and cardiac remodeling, specifically with hypertrophy (r = 0.51, P≤0.01), myocardial collagen (r = 0.40, P≤0.05) and ED1 immunostaining (r = 0.48, P≤0.01). In the untreated hypertensive group, increased levels (P<0.05) of the proinflammatory molecules p65 NF-κB, vascular cell adhesion molecule 1 and interleukin-6 antibody in the myocardium, aortic wall and PBMC were observed and were reduced with fasudil (P<0.05). In conclusion, in this hypertension model, Rho-kinase and its pathway activation determined in circulating leukocytes reflect the activation of this pathway in the myocardium and in the aortic wall and are significantly related to myocardial remodeling (hypertrophy, fibrosis and inflammation).

Abstract MP09: Vascular Rho-kinase Activation by Inflammatory Signaling Mediates Blood Pressure Increase in Overweight

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Cristiane Aoqui Aoqui ◽  
Stefan Chmielewski ◽  
Uwe Heemann ◽  
Marcus Baumann
Keyword(s):  
Blood Pressure ◽  
Kinase Activity ◽  
Rho Kinase ◽  
Normal Weight ◽  
Mesenteric Arteries ◽  
Pressure Increase ◽  
Tnf Alpha ◽  
Kinase Activation ◽  
Blood Pressure Increase ◽  
Tlr4 Ligand

Background: Overweight is associated with a high prevalence of hypertension. The mechanisms linking overweight to blood pressure increase remain unclear. We hypothesized that vascular Rho-kinase activation contributes to blood pressure increase in overweight by involving TNF-α and TLR4. Methods: C57/BL6 mice fed a high-fat diet for 2 weeks were used to induce overweight associated blood pressure increase. Additional treatment in overweight and normal weight mice contained Rho-kinase inhibitors (fasudil and pravastatin; n=7/all groups), etanercept and TNFR1 and TLR4 null-mice. Microvascular studies were performed in a wire myograph and arterial blood pressure measured with a carotid catheter. Rho-kinase activity was determined in small mesenteric arteries of all groups. Inflammatory ligands such as TNF-alpha and free fatty acids were determined. Effects of TNF-alpha and TLR4 ligand LPS and palmitic acid on Rho-kinase activity and were determined ex vivo in mesenteric arteries and in in vivo. Results: Overweight mice had higher blood pressure (Delta: 9±2 mmHg) and vasoconstriction as normal weight mice. Small mesenteric arteries of overweight mice had a 50% higher Rho-kinase activity as normal weight mice. Ex vivo treatment with the Rho-kinase inhibitor Y-27632 reversed vasoconstriction of mesenteric arteries of overweight mice to constriction level in normal weight mice. In vivo treatment with the Rho-kinase inhibitor fasudil or pravastatin along with high-fat diet abolished the overweight associated blood pressure increase and enhanced vasoconstriction. TNF-alpha and TLR4 ligand free fatty acid were enhanced in overweight mice. TNF-alpha and TLR4 ligand LPS and palmitic acid increased Rho-kinase activity in mesenteric arteries. Use of etenercept, TNFR1 and TLR4 null-mice in the overweight model prevented blood pressure increase, vasoconstriction and Rho-kinase activation. All described effects were independent of adiposity. Conclusion: These results indicated that in diet-induced overweight vascular Rho-kinase activation is a key element of increased blood pressure and vasoconstriction. Potential activator of Rho-kinase are mediated by inflammatory factors including TLR4 ligands and TNF-alpha.

scholarly journals Diuretics prevent Rho-kinase activation and expression of profibrotic/oxidative genes in the hypertensive aortic wall

2016 ◽  
Vol 10 (6) ◽  
pp. 338-347 ◽  
Author(s):  
Patricio Araos ◽  
David Mondaca ◽  
Jorge E. Jalil ◽  
Cristián Yañez ◽  
Ulises Novoa ◽  
...  
Keyword(s):  
Gene Expression ◽  
Blood Pressure ◽  
Growth Factor ◽  
Aortic Wall ◽  
Transforming Growth Factor ◽  
Rho Kinase ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Tissue Growth ◽  
Hypertensive Rats ◽  
Rock Inhibitor

Background: Diuretics are current antihypertensive drugs since they reduce blood pressure and cardiovascular risk. Increased vascular tone is modulated in a relevant way by the RhoA/Rho-kinase (ROCK) pathway, by acting on vascular smooth muscle cell contraction. This pathway has also proremodeling vascular effects. There are few data on the role of diuretics on both vascular ROCK activation and on proremodeling effects. We assessed the effects of hydrochlorothiazide (HCTZ) and spironolactone (spiro) alone and in combination with the ROCK inhibitor fasudil (FAS) on ROCK activation, gene expression of proremodeling markers and on hypertrophy in the aortic wall of hypertensive rats. Methods: Deoxycorticosterone acetate (DOCA)-salt hypertensive rats (male, Sprague–Dawley) were randomized to the specific ROCK inhibitor FAS, HCTZ, spiro or the combinations of FAS/HCTZ or FAS/spiro for 3 weeks. At the end of the study, ROCK activation (by western blot), gene expression of proremodeling markers (by reverse transcription polymerase chain reaction, RT-PCR) and vascular hypertrophy (by morphometry) were determined in the aortic wall. Results: All treatments significantly reduced blood pressure. In the DOCA rats the p-myosin phosphatase target protein-1 (MYPT1)/t-MYPT1 ratio, index of ROCK activation was higher by 2.8 fold ( p < 0.05) compared with control rats. All treatments reduced ROCK activation in the aortic wall to control levels ( p < 0.05). Besides, significantly increased protein levels of transforming growth factor β1 (TGF-β1), gene expression of TGF-β1, connective tissue growth factor (CTGF), p22 phox and gp91 phox subunits of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, as well as increased media thickness and aortic media area/lumen area (AM/LA) in the untreated hypertensive rats were significantly reduced ( p < 0.05) to control levels by all treatments. Similar effects were observed using both diuretics alone or in combination with FAS. Conclusions: In the aortic wall, both HCTZ and spiro in antihypertensive doses reduce ROCK activation, subsequent expression of genes that promote vascular remodeling and hypertrophy in this experimental model of hypertension. These effects could explain some of their clinical benefits in hypertensive patients.

Effect of Early Normotension with Olmesartan on Rho-kinase Activity in Hypertensive Patients

2019 ◽  
Vol 18 (1) ◽  
pp. 87-91 ◽  
Author(s):  
Claudio Cantin ◽  
Jorge E. Jalil ◽  
Juan F. Bulnes ◽  
Ulises Novoa ◽  
Paul MacNab ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Kinase Activity ◽  
Clinical Evidence ◽  
Mononuclear Cells ◽  
Treatment Initiation ◽  
Rho Kinase ◽  
Angiotensin Ii Receptor ◽  
Peripheral Blood Mononuclear ◽  
Hypertensive Patients ◽  
Rock Activity

Background: Angiotensin II is a potent activator of the Rho-kinase (ROCK) pathway, through which it exerts some of its adverse vasoconstrictor effects. Clinical evidence on the effects of blocking the angiotensin II receptor 1 on ROCK activity in hypertensive patients is scarce. Objective: To demonstrate that ROCK activity in peripheral blood mononuclear cells (PMBCs) in patients with essential hypertension is reduced earlier than previously observed, along with blood pressure (BP) lowering on treatment with olmesartan. Methods: Prospective pilot open study; 17 hypertensive patients were treated with progressive olmesartan doses starting with 20 mg qd. BP was measured at 3, 6 and 9 weeks after treatment initiation. If treatment failed to normalize BP after 3 weeks, olmesartan dose was increased to 40 mg qd, and if still hypertensive after 6 weeks, 12.5 mg of hydrochlorothiazide qd was added. ROCK activity was measured at baseline and 9 weeks after treatment as myosin phosphatase target subunit 1 phosphorylation (MYPT1-p/T ratio) in PBMC. Results: Mean baseline BP was 162 ± 4.9/101 ± 2.4 mmHg. After 9 weeks of treatment, both systolic and diastolic BP were reduced by 41 and 22 mmHg, respectively (p<0.05). Mean pretreatment MYPT1- p/T ratio in PMBCs was significantly reduced by 80% after 9 weeks with olmesartan (p<0.01). Conclusion: Normotension achieved after 9 weeks in 82% of the patients treated with olmesartan was associated with a significant reduction of ROCK activity in PBMC.

scholarly journals Pro-Nerve Growth Factor Induces Activation of RhoA Kinase and Neuronal Cell Death

2019 ◽  
Vol 9 (8) ◽  
pp. 204 ◽  
Author(s):  
Marina Sycheva ◽  
Jake Sustarich ◽  
Yuxian Zhang ◽  
Vaithinathan Selvaraju ◽  
Thangiah Geetha ◽  
...  
Keyword(s):  
Cell Death ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Kinase Activity ◽  
Kinase Inhibitor ◽  
Neuronal Cell Death ◽  
Rho Kinase ◽  
Neuronal Cell ◽  
Nerve Growth ◽  
Rhoa Kinase

We have previously shown that the expression of pro-nerve growth factor (proNGF) was significantly increased, nerve growth factor (NGF) level was decreased, and the expression of p75NTR was enhanced in Alzheimer’s disease (AD) hippocampal samples. NGF regulates cell survival and differentiation by binding TrkA and p75NTR receptors. ProNGF is the precursor form of NGF, binds to p75NTR, and induces cell apoptosis. The objective of this study is to determine whether the increased p75NTR expression in AD is due to the accumulation of proNGF and Rho kinase activation. PC12 cells were stimulated with either proNGF or NGF. Pull-down assay was carried out to determine the RhoA kinase activity. We found the expression of p75NTR was enhanced by proNGF compared to NGF. The proNGF stimulation also increased the RhoA kinase activity leading to apoptosis. The expression of active RhoA kinase was found to be increased in human AD hippocampus compared to control. The addition of RhoA kinase inhibitor Y27632 not only blocked the RhoA kinase activity but also reduced the expression of p75NTR receptor and inhibited the activation of JNK and MAPK induced by proNGF. This suggests that overexpression of proNGF in AD enhances p75NTR expression and activation of RhoA, leading to neuronal cell death.

Region-Specific Medial Fiber Micro-Architecture in the Longitudinal-Radial and Circumferential-Radial Planes of Ascending Thoracic Aortic Aneurysm Among Bicuspid and Tricuspid Aortic Valve Patients

2013 ◽  
Author(s):  
Alkiviadis Tsamis ◽  
Julie A. Phillippi ◽  
Ryan G. Koch ◽  
Jeffrey T. Krawiec ◽  
Antonio D’Amore ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Aortic Valve ◽  
Aortic Aneurysm ◽  
Thoracic Aortic Aneurysm ◽  
Aortic Wall ◽  
False Lumen ◽  
True Lumen ◽  
Life Threatening ◽  
The Media ◽  
Intimal Tear

Aortic dissection is a life-threatening cardiovascular emergency with a high potential for death. It usually begins with an intimal tear which permits blood to enter the wall, split the media and create a false lumen, which can reenter the true lumen or exit through the adventitia causing complete rupture. A possible mechanism for dissection of ascending thoracic aortic aneurysm (ATAA) can be the occurrence of blood pressure-induced wall stresses in excess to the adhesive strength between the degenerated aortic wall layers.

Antihypertensive Property and Renal Protection by Shichimotsu-koka-to Extract in Salt-induced Hypertension in Dahl Strain Rats

1994 ◽  
Vol 22 (01) ◽  
pp. 51-62 ◽  
Author(s):  
Nobuhito Hiwara ◽  
Yoshio Uehara ◽  
Satoru Takada ◽  
Yukari Kawabata ◽  
Nobuko Ohshima ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Renal Function ◽  
Aortic Wall ◽  
Antihypertensive Effect ◽  
Blood Pressure Reduction ◽  
Glomerular Sclerosis ◽  
Pressure Reduction ◽  
Renal Protection ◽  
Induced Hypertension ◽  
Sclerotic Lesions

We determined whether or not the kampo formula, Shichimotsu-koka-to extract, attenuates the development of salt-induced hypertension and provides renal protection against hypertensive injury in Dahl salt-sensitive (Dahl S) rats. A six-week treatment using this formula dose-dependently decreased the systolic blood pressure in Dahl S rats fed a high-salt (2% NaCl) diet. This blood pressure reduction was associated with a decrease in the thickness of the aortic wall. Renal function was not altered with this treatment; however, glomerular sclerotic lesions in the kidney were significantly attenuated. Neither arterial nor tubular lesions were affected. These data suggest that Shichimotsu-koka-to extract exhibits an antihypertensive effect which is associated with partial resolution of glomerular sclerosis in the kidney.

scholarly journals Activation of G protein-coupled bile acid receptor, TGR5, induces smooth muscle relaxation via both Epac- and PKA-mediated inhibition of RhoA/Rho kinase pathway

2013 ◽  
Vol 304 (5) ◽  
pp. G527-G535 ◽  
Author(s):  
Senthilkumar Rajagopal ◽  
Divya P. Kumar ◽  
Sunila Mahavadi ◽  
Sayak Bhattacharya ◽  
Ruizhe Zhou ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Kinase Activity ◽  
Muscle Relaxation ◽  
Rho Kinase ◽  
Muscle Cells ◽  
Receptor Activation ◽  
Gastric Smooth Muscle ◽  
Selective Ligand ◽  
Gastric Muscle ◽  
Kinase Pathway

The present study characterized the TGR5 expression and the signaling pathways coupled to this receptor that mediates the relaxation of gastric smooth muscle. TGR5 was detected in gastric muscle cells by RT-PCR and Western blotting. Treatment of cells with the TGR5-selective ligand oleanolic acid (OA) activated Gαs, but not Gαq, Gαi1, Gαi2, or Gαi3, and increased cAMP levels. OA did not elicit contraction, but caused relaxation of carbachol-induced contraction of gastric muscle cells from wild-type mice, but not tgr5−/− mice. OA, but not a selective exchange protein activated by cAMP (Epac) ligand (8-pCPT-2′-O-Me-cAMP), caused phosphorylation of RhoA and the phosphorylation was blocked by the PKA inhibitor, myristoylated PKI, and by the expression of phosphorylation-deficient mutant RhoA (S188A). Both OA and Epac ligand stimulated Ras-related protein 1 (Rap1) and inhibited carbachol (CCh)-induced Rho kinase activity. Expression of RhoA (S188A) or PKI partly reversed the inhibition of Rho kinase activity by OA but had no effect on inhibition by Epac ligand. However, suppression of Rap1 with siRNA blocked the inhibition of Rho kinase by Epac ligand, and partly reversed the inhibition by OA; the residual inhibition was blocked by PKI. Muscle relaxation in response to OA, but not Epac ligand, was partly reversed by PKI. We conclude that activation of TGR5 causes relaxation of gastric smooth muscle and the relaxation is mediated through inhibition of RhoA/Rho kinase pathway via both cAMP/Epac-dependent stimulation of Rap1 and cAMP/PKA-dependent phosphorylation of RhoA at Ser188. TGR5 receptor activation on smooth muscle reveals a novel mechanism for the regulation of gut motility by bile acids.

scholarly journals Effects of Tolvaptan on Oxidative Stress in ADPKD: A Molecular Biological Approach

2022 ◽  
Vol 11 (2) ◽  
pp. 402
Author(s):  
Matteo Rigato ◽  
Gianni Carraro ◽  
Irene Cirella ◽  
Silvia Dian ◽  
Valentina Di Di Vico ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Protein Expression ◽  
Molecular Mechanisms ◽  
Kinase Activity ◽  
Rho Kinase ◽  
Renal Deterioration ◽  
Deterioration Rate ◽  
Function Loss ◽  
State Marker ◽  
Molecular Biological Approach

Autosomal dominant polycystic disease (ADPKD) is the most frequent monogenic kidney disease. It causes progressive renal failure, endothelial dysfunction, and hypertension, all of which are strictly linked to oxidative stress (OxSt). Treatment with tolvaptan is known to slow the renal deterioration rate, but not all the molecular mechanisms involved in this effect are well-established. We evaluated the OxSt state in untreated ADPKD patients compared to that in tolvaptan-treated ADPKD patients and healthy subjects. OxSt was assessed in nine patients for each group in terms of mononuclear cell p22phox protein expression, NADPH oxidase key subunit, MYPT-1 phosphorylation state, marker of Rho kinase activity (Western blot) and heme oxygenase (HO)-1, induced and protective against OxSt (ELISA). p22phox protein expression was higher in untreated ADPKD patients compared to treated patients and controls: 1.42 ± 0.11 vs. 0.86 ± 0.15 d.u., p = 0.015, vs. 0.53 ± 0.11 d.u., p < 0.001, respectively. The same was observed for phosphorylated MYPT-1: 0.96 ± 0.28 vs. 0.68 ± 0.09 d.u., p = 0.013 and vs. 0.47 ± 0.13 d.u., p < 0.001, respectively, while the HO-1 expression of untreated patients was significantly lower compared to that of treated patients and controls: 5.33 ± 3.34 vs. 2.08 ± 0.79 ng/mL, p = 0.012, vs. 1.97 ± 1.22 ng/mL, p = 0.012, respectively. Tolvaptan-treated ADPKD patients have reduced OxSt levels compared to untreated patients. This effect may contribute to the slowing of renal function loss observed with tolvaptan treatment.

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