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An interaction of renin-angiotensin and kallikrein-kinin systems contributes to vascular hypertrophy in angiotensin II-induced hypertension: in vivo and in vitro studies.

Ceravolo GS, Montezano AC, Jordão MT, Akamine EH, Costa TJ, Takano AP, Fernandes DC, Barreto-Chaves ML, Laurindo FR, Tostes RC, Fortes ZB, Chopard RP, Touyz RM, Carvalho MH - PLoS ONE (2014)

Bottom Line: At this concentration, neither ANG II nor the B1R agonist produced any effects when tested individually.This is associated with activation of redox-regulated ERK1/2 pathway that controls aortic smooth muscle cells growth.Our findings highlight an important cross-talk between the DABK and ANG II in the vascular system and contribute to a better understanding of the mechanisms involved in vascular remodeling in hypertension.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, Brazil; Department of Physiological Sciences, Biological Sciences Center, State University of Londrina, Londrina, Brazil.

ABSTRACT
The kallikrein-kinin and renin-angiotensin systems interact at multiple levels. In the present study, we tested the hypothesis that the B1 kinin receptor (B1R) contributes to vascular hypertrophy in angiotensin II (ANG II)-induced hypertension, through a mechanism involving reactive oxygen species (ROS) generation and extracellular signal-regulated kinase (ERK1/2) activation. Male Wistar rats were infused with vehicle (control rats), 400 ng/Kg/min ANG II (ANG II rats) or 400 ng/Kg/min ANG II plus B1 receptor antagonist, 350 ng/Kg/min des-Arg(9)-Leu(8)-bradykinin (ANGII+DAL rats), via osmotic mini-pumps (14 days) or received ANG II plus losartan (10 mg/Kg, 14 days, gavage - ANG II+LOS rats). After 14 days, ANG II rats exhibited increased systolic arterial pressure [(mmHg) 184 ± 5.9 vs 115 ± 2.3], aortic hypertrophy; increased ROS generation [2-hydroxyethidium/dihydroethidium (EOH/DHE): 21.8 ± 2.7 vs 6.0 ± 1.8] and ERK1/2 phosphorylation (% of control: 218.3 ± 29.4 vs 100 ± 0.25]. B1R expression was increased in aortas from ANG II and ANG II+DAL rats than in aortas from the ANG II+LOS and control groups. B1R antagonism reduced aorta hypertrophy, prevented ROS generation (EOH/DHE: 9.17 ± 3.1) and ERK1/2 phosphorylation (137 ± 20.7%) in ANG II rats. Cultured aortic vascular smooth muscle cells (VSMC) stimulated with low concentrations (0.1 nM) of ANG II plus B1R agonist exhibited increased ROS generation, ERK1/2 phosphorylation, proliferating-cell nuclear antigen expression and [H3]leucine incorporation. At this concentration, neither ANG II nor the B1R agonist produced any effects when tested individually. The ANG II/B1R agonist synergism was inhibited by losartan (AT1 blocker, 10 µM), B1R antagonist (10 µM) and Tiron (superoxide anion scavenger, 10 mM). These data suggest that B1R activation contributes to ANG II-induced aortic hypertrophy. This is associated with activation of redox-regulated ERK1/2 pathway that controls aortic smooth muscle cells growth. Our findings highlight an important cross-talk between the DABK and ANG II in the vascular system and contribute to a better understanding of the mechanisms involved in vascular remodeling in hypertension.

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Effects of des-Arg9-bradykinin (DABK) and angiotensin II (ANG II) vascular smooth muscle cells (VSMC) growth.[A] Aortic VSMC from Wistar rats treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM) and TIRON (1 mM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are represented as mean±SEM of 4 experiments. *P<0.05 vs Vehicle. [B] Aortic VSMC from Wistar rats with ANG II at high concentration (100 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are expressed as mean±SEM of 5 experiments. *P<0.05 vs vehicle. [C] [H3] leucine incorporation in VSMC treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM) and des-arg9-leu8-bradykinin (DAL – 10 µM) and VSMC treated with ANG II at high concentration (100 nM). Results are expressed as mean±SEM of 3 experiments and *P<0.05 vs vehicle.
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pone-0111117-g005: Effects of des-Arg9-bradykinin (DABK) and angiotensin II (ANG II) vascular smooth muscle cells (VSMC) growth.[A] Aortic VSMC from Wistar rats treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM) and TIRON (1 mM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are represented as mean±SEM of 4 experiments. *P<0.05 vs Vehicle. [B] Aortic VSMC from Wistar rats with ANG II at high concentration (100 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are expressed as mean±SEM of 5 experiments. *P<0.05 vs vehicle. [C] [H3] leucine incorporation in VSMC treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM) and des-arg9-leu8-bradykinin (DAL – 10 µM) and VSMC treated with ANG II at high concentration (100 nM). Results are expressed as mean±SEM of 3 experiments and *P<0.05 vs vehicle.

Mentions: PCNA expression and [H3] leucine incorporation were assessed as molecular markers of cell growth. As demonstrated in Figure 5A, ANG II and DABK at low concentration (0.1 nM), increased PCNA expression only when added together and these effects were abolished by LOS, DAL and Tiron. In the absence of DABK, ANG II at high concentration (100 nM) increased PCNA expression when compared with vehicle. This effect was not changed by DAL, a B1R antagonist, but was inhibited by LOS, AT1 receptor antagonist (Figure 5B). ANG II and DABK at low concentration (0.1 nM) increased [H3] leucine incorporation only when added together and LOS plus DAL abolished this effect. ANG II at high concentration (100 nM) increased [H3] leucine incorporation when compared with the control (vehicle) cells (Figure 5C).


An interaction of renin-angiotensin and kallikrein-kinin systems contributes to vascular hypertrophy in angiotensin II-induced hypertension: in vivo and in vitro studies.

Ceravolo GS, Montezano AC, Jordão MT, Akamine EH, Costa TJ, Takano AP, Fernandes DC, Barreto-Chaves ML, Laurindo FR, Tostes RC, Fortes ZB, Chopard RP, Touyz RM, Carvalho MH - PLoS ONE (2014)

Effects of des-Arg9-bradykinin (DABK) and angiotensin II (ANG II) vascular smooth muscle cells (VSMC) growth.[A] Aortic VSMC from Wistar rats treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM) and TIRON (1 mM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are represented as mean±SEM of 4 experiments. *P<0.05 vs Vehicle. [B] Aortic VSMC from Wistar rats with ANG II at high concentration (100 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are expressed as mean±SEM of 5 experiments. *P<0.05 vs vehicle. [C] [H3] leucine incorporation in VSMC treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM) and des-arg9-leu8-bradykinin (DAL – 10 µM) and VSMC treated with ANG II at high concentration (100 nM). Results are expressed as mean±SEM of 3 experiments and *P<0.05 vs vehicle.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4219703&req=5

pone-0111117-g005: Effects of des-Arg9-bradykinin (DABK) and angiotensin II (ANG II) vascular smooth muscle cells (VSMC) growth.[A] Aortic VSMC from Wistar rats treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM) and TIRON (1 mM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are represented as mean±SEM of 4 experiments. *P<0.05 vs Vehicle. [B] Aortic VSMC from Wistar rats with ANG II at high concentration (100 nM) in the presence or not of losartan (LOS – 10 µM), des-arg9-leu8-bradykinin (DAL – 10 µM). Bar graph shows PCNA expression normalized against the housekeeping protein β-actin. Results are expressed as mean±SEM of 5 experiments. *P<0.05 vs vehicle. [C] [H3] leucine incorporation in VSMC treated with ANG II and/or DABK at low concentration (0.1 nM) in the presence or not of losartan (LOS – 10 µM) and des-arg9-leu8-bradykinin (DAL – 10 µM) and VSMC treated with ANG II at high concentration (100 nM). Results are expressed as mean±SEM of 3 experiments and *P<0.05 vs vehicle.
Mentions: PCNA expression and [H3] leucine incorporation were assessed as molecular markers of cell growth. As demonstrated in Figure 5A, ANG II and DABK at low concentration (0.1 nM), increased PCNA expression only when added together and these effects were abolished by LOS, DAL and Tiron. In the absence of DABK, ANG II at high concentration (100 nM) increased PCNA expression when compared with vehicle. This effect was not changed by DAL, a B1R antagonist, but was inhibited by LOS, AT1 receptor antagonist (Figure 5B). ANG II and DABK at low concentration (0.1 nM) increased [H3] leucine incorporation only when added together and LOS plus DAL abolished this effect. ANG II at high concentration (100 nM) increased [H3] leucine incorporation when compared with the control (vehicle) cells (Figure 5C).

Bottom Line: At this concentration, neither ANG II nor the B1R agonist produced any effects when tested individually.This is associated with activation of redox-regulated ERK1/2 pathway that controls aortic smooth muscle cells growth.Our findings highlight an important cross-talk between the DABK and ANG II in the vascular system and contribute to a better understanding of the mechanisms involved in vascular remodeling in hypertension.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, Brazil; Department of Physiological Sciences, Biological Sciences Center, State University of Londrina, Londrina, Brazil.

ABSTRACT
The kallikrein-kinin and renin-angiotensin systems interact at multiple levels. In the present study, we tested the hypothesis that the B1 kinin receptor (B1R) contributes to vascular hypertrophy in angiotensin II (ANG II)-induced hypertension, through a mechanism involving reactive oxygen species (ROS) generation and extracellular signal-regulated kinase (ERK1/2) activation. Male Wistar rats were infused with vehicle (control rats), 400 ng/Kg/min ANG II (ANG II rats) or 400 ng/Kg/min ANG II plus B1 receptor antagonist, 350 ng/Kg/min des-Arg(9)-Leu(8)-bradykinin (ANGII+DAL rats), via osmotic mini-pumps (14 days) or received ANG II plus losartan (10 mg/Kg, 14 days, gavage - ANG II+LOS rats). After 14 days, ANG II rats exhibited increased systolic arterial pressure [(mmHg) 184 ± 5.9 vs 115 ± 2.3], aortic hypertrophy; increased ROS generation [2-hydroxyethidium/dihydroethidium (EOH/DHE): 21.8 ± 2.7 vs 6.0 ± 1.8] and ERK1/2 phosphorylation (% of control: 218.3 ± 29.4 vs 100 ± 0.25]. B1R expression was increased in aortas from ANG II and ANG II+DAL rats than in aortas from the ANG II+LOS and control groups. B1R antagonism reduced aorta hypertrophy, prevented ROS generation (EOH/DHE: 9.17 ± 3.1) and ERK1/2 phosphorylation (137 ± 20.7%) in ANG II rats. Cultured aortic vascular smooth muscle cells (VSMC) stimulated with low concentrations (0.1 nM) of ANG II plus B1R agonist exhibited increased ROS generation, ERK1/2 phosphorylation, proliferating-cell nuclear antigen expression and [H3]leucine incorporation. At this concentration, neither ANG II nor the B1R agonist produced any effects when tested individually. The ANG II/B1R agonist synergism was inhibited by losartan (AT1 blocker, 10 µM), B1R antagonist (10 µM) and Tiron (superoxide anion scavenger, 10 mM). These data suggest that B1R activation contributes to ANG II-induced aortic hypertrophy. This is associated with activation of redox-regulated ERK1/2 pathway that controls aortic smooth muscle cells growth. Our findings highlight an important cross-talk between the DABK and ANG II in the vascular system and contribute to a better understanding of the mechanisms involved in vascular remodeling in hypertension.

Show MeSH
Related in: MedlinePlus