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Increased migration of monocytes in essential hypertension is associated with increased transient receptor potential channel canonical type 3 channels.

Zhao Z, Ni Y, Chen J, Zhong J, Yu H, Xu X, He H, Yan Z, Scholze A, Liu D, Zhu Z, Tepel M - PLoS ONE (2012)

Bottom Line: Proteins were identified by immunoblotting and quantitative in-cell Western assay.The effects of TRP channel-inhibitor 2-aminoethoxydiphenylborane (2-APB) and small interfering RNA knockdown of TRPC3 were investigated.The fMLP-induced changes of cytosolic calcium were significantly increased in monocytes from hypertensive patients compared to normotensive control subjects.

View Article: PubMed Central - PubMed

Affiliation: Department of Hypertension and Endocrinology, Center for Hypertension and Metabolic Diseases, Daping Hospital, Third Military Medical University, Chongqing Institute of Hypertension, Chongqing, China.

ABSTRACT
Increased transient receptor potential canonical type 3 (TRPC3) channels have been observed in patients with essential hypertension. In the present study we tested the hypothesis that increased monocyte migration is associated with increased TRPC3 expression. Monocyte migration assay was performed in a microchemotaxis chamber using chemoattractants formylated peptide Met-Leu-Phe (fMLP) and tumor necrosis factor-α (TNF-α). Proteins were identified by immunoblotting and quantitative in-cell Western assay. The effects of TRP channel-inhibitor 2-aminoethoxydiphenylborane (2-APB) and small interfering RNA knockdown of TRPC3 were investigated. We observed an increased fMLP-induced migration of monocytes from hypertensive patients compared with normotensive control subjects (246 ± 14% vs 151 ± 10%). The TNF-α-induced migration of monocytes in patients with essential hypertension was also significantly increased compared to normotensive control subjects (221 ± 20% vs 138 ± 18%). In the presence of 2-APB or after siRNA knockdown of TRPC3 the fMLP-induced monocyte migration was significantly blocked. The fMLP-induced changes of cytosolic calcium were significantly increased in monocytes from hypertensive patients compared to normotensive control subjects. The fMLP-induced monocyte migration was significantly reduced in the presence of inhibitors of tyrosine kinase and phosphoinositide 3-kinase. We conclude that increased monocyte migration in patients with essential hypertension is associated with increased TRPC3 channels.

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The role of Akt and ERK-dependent pathways in essential hypertension.A, B; fMLP activates ERK or phosphorylation of ERK (A) and Akt or phosphorylation of Akt (B) in a dose- and time-dependent manner in monocytes from normotensive control subjects. 10 nmol/L open bars, 100 nmol/L filled bars. Data are mean ± SEM, n = 3. *p<0.05 compared to lower concentration conditions. C, D; Increased fMLP-induced phosphorylation of ERK (C) and Akt (D) in monocytes from patients with essential hypertension. The proteins were measured using immunoblotting with specific antibodies. Data are mean ± SEM from three independent experiments. *p<0.05 compared to normotensive control subjects. E; fMLP activates monocytes by an ERK-dependent and Akt-dependent pathway. Akt, ERK, or pERK and pAkt were measured using immunoblotting with specific antibodies. In the presence of 2-APB or after administration of specific siRNA against TRPC3, the fMLP-induced ERK, pERK; Akt and pAkt were significantly reduced when compared with control conditions. Data are mean ± SEM from six independent experiments. *p<0.05; **p<0.01 compared to control.
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pone-0032628-g006: The role of Akt and ERK-dependent pathways in essential hypertension.A, B; fMLP activates ERK or phosphorylation of ERK (A) and Akt or phosphorylation of Akt (B) in a dose- and time-dependent manner in monocytes from normotensive control subjects. 10 nmol/L open bars, 100 nmol/L filled bars. Data are mean ± SEM, n = 3. *p<0.05 compared to lower concentration conditions. C, D; Increased fMLP-induced phosphorylation of ERK (C) and Akt (D) in monocytes from patients with essential hypertension. The proteins were measured using immunoblotting with specific antibodies. Data are mean ± SEM from three independent experiments. *p<0.05 compared to normotensive control subjects. E; fMLP activates monocytes by an ERK-dependent and Akt-dependent pathway. Akt, ERK, or pERK and pAkt were measured using immunoblotting with specific antibodies. In the presence of 2-APB or after administration of specific siRNA against TRPC3, the fMLP-induced ERK, pERK; Akt and pAkt were significantly reduced when compared with control conditions. Data are mean ± SEM from six independent experiments. *p<0.05; **p<0.01 compared to control.

Mentions: We observed that fMLP activates monocytes by an ERK- and Akt-dependent pathway. As shown in Figure 6, administration of fMLP significantly increased phosphorylated ERK and phosphorylated Akt in a dose-dependent and time-dependent manner (Figure 6A and 6B). Furthermore, we compared the dose response effects of fMLP on monocytes from patients with essential hypertension and normotensive control subjects. We observed an increased phosphorylated ERK (Figure 6C) and phosphorylated Akt (Figure 6D) after fMLP stimulation of monocytes from patients with essential hypertension compared to normotensive control subjects. The expression of pERK was 2.77±0.26 vs. 1.55±0.06, n = 3, P<0.05; and the expression of pAkt was 0.56±0.04 vs. 0.24±0.04, n = 3, P<0.05 for these groups, respectively. These findings confirmed that the fMLP-induced activation of monocytes in patients with essential hypertension was associated with ERK and Akt pathways.


Increased migration of monocytes in essential hypertension is associated with increased transient receptor potential channel canonical type 3 channels.

Zhao Z, Ni Y, Chen J, Zhong J, Yu H, Xu X, He H, Yan Z, Scholze A, Liu D, Zhu Z, Tepel M - PLoS ONE (2012)

The role of Akt and ERK-dependent pathways in essential hypertension.A, B; fMLP activates ERK or phosphorylation of ERK (A) and Akt or phosphorylation of Akt (B) in a dose- and time-dependent manner in monocytes from normotensive control subjects. 10 nmol/L open bars, 100 nmol/L filled bars. Data are mean ± SEM, n = 3. *p<0.05 compared to lower concentration conditions. C, D; Increased fMLP-induced phosphorylation of ERK (C) and Akt (D) in monocytes from patients with essential hypertension. The proteins were measured using immunoblotting with specific antibodies. Data are mean ± SEM from three independent experiments. *p<0.05 compared to normotensive control subjects. E; fMLP activates monocytes by an ERK-dependent and Akt-dependent pathway. Akt, ERK, or pERK and pAkt were measured using immunoblotting with specific antibodies. In the presence of 2-APB or after administration of specific siRNA against TRPC3, the fMLP-induced ERK, pERK; Akt and pAkt were significantly reduced when compared with control conditions. Data are mean ± SEM from six independent experiments. *p<0.05; **p<0.01 compared to control.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3306381&req=5

pone-0032628-g006: The role of Akt and ERK-dependent pathways in essential hypertension.A, B; fMLP activates ERK or phosphorylation of ERK (A) and Akt or phosphorylation of Akt (B) in a dose- and time-dependent manner in monocytes from normotensive control subjects. 10 nmol/L open bars, 100 nmol/L filled bars. Data are mean ± SEM, n = 3. *p<0.05 compared to lower concentration conditions. C, D; Increased fMLP-induced phosphorylation of ERK (C) and Akt (D) in monocytes from patients with essential hypertension. The proteins were measured using immunoblotting with specific antibodies. Data are mean ± SEM from three independent experiments. *p<0.05 compared to normotensive control subjects. E; fMLP activates monocytes by an ERK-dependent and Akt-dependent pathway. Akt, ERK, or pERK and pAkt were measured using immunoblotting with specific antibodies. In the presence of 2-APB or after administration of specific siRNA against TRPC3, the fMLP-induced ERK, pERK; Akt and pAkt were significantly reduced when compared with control conditions. Data are mean ± SEM from six independent experiments. *p<0.05; **p<0.01 compared to control.
Mentions: We observed that fMLP activates monocytes by an ERK- and Akt-dependent pathway. As shown in Figure 6, administration of fMLP significantly increased phosphorylated ERK and phosphorylated Akt in a dose-dependent and time-dependent manner (Figure 6A and 6B). Furthermore, we compared the dose response effects of fMLP on monocytes from patients with essential hypertension and normotensive control subjects. We observed an increased phosphorylated ERK (Figure 6C) and phosphorylated Akt (Figure 6D) after fMLP stimulation of monocytes from patients with essential hypertension compared to normotensive control subjects. The expression of pERK was 2.77±0.26 vs. 1.55±0.06, n = 3, P<0.05; and the expression of pAkt was 0.56±0.04 vs. 0.24±0.04, n = 3, P<0.05 for these groups, respectively. These findings confirmed that the fMLP-induced activation of monocytes in patients with essential hypertension was associated with ERK and Akt pathways.

Bottom Line: Proteins were identified by immunoblotting and quantitative in-cell Western assay.The effects of TRP channel-inhibitor 2-aminoethoxydiphenylborane (2-APB) and small interfering RNA knockdown of TRPC3 were investigated.The fMLP-induced changes of cytosolic calcium were significantly increased in monocytes from hypertensive patients compared to normotensive control subjects.

View Article: PubMed Central - PubMed

Affiliation: Department of Hypertension and Endocrinology, Center for Hypertension and Metabolic Diseases, Daping Hospital, Third Military Medical University, Chongqing Institute of Hypertension, Chongqing, China.

ABSTRACT
Increased transient receptor potential canonical type 3 (TRPC3) channels have been observed in patients with essential hypertension. In the present study we tested the hypothesis that increased monocyte migration is associated with increased TRPC3 expression. Monocyte migration assay was performed in a microchemotaxis chamber using chemoattractants formylated peptide Met-Leu-Phe (fMLP) and tumor necrosis factor-α (TNF-α). Proteins were identified by immunoblotting and quantitative in-cell Western assay. The effects of TRP channel-inhibitor 2-aminoethoxydiphenylborane (2-APB) and small interfering RNA knockdown of TRPC3 were investigated. We observed an increased fMLP-induced migration of monocytes from hypertensive patients compared with normotensive control subjects (246 ± 14% vs 151 ± 10%). The TNF-α-induced migration of monocytes in patients with essential hypertension was also significantly increased compared to normotensive control subjects (221 ± 20% vs 138 ± 18%). In the presence of 2-APB or after siRNA knockdown of TRPC3 the fMLP-induced monocyte migration was significantly blocked. The fMLP-induced changes of cytosolic calcium were significantly increased in monocytes from hypertensive patients compared to normotensive control subjects. The fMLP-induced monocyte migration was significantly reduced in the presence of inhibitors of tyrosine kinase and phosphoinositide 3-kinase. We conclude that increased monocyte migration in patients with essential hypertension is associated with increased TRPC3 channels.

Show MeSH
Related in: MedlinePlus