Limits...
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.

Show MeSH

Related in: MedlinePlus

Monocyte subtypes and fMLP receptors in normotensive and hypertensive patients.A, Peripheral blood monocytes subpopulations were analyzed by flow-cytometry. After labeling with anti-CD14 phycoerythrin (PE) conjugated and anti-CD16 FITC conjugated, monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars) were readily separated into three distinct subsets according to CD14 and CD16 positivity. Data are mean ± SEM, each n = 11, P>0.05 NT vs. HT. B, Expression of fMLP receptors using immunoblotting with specific antibodies. The data showed that fMLP receptors were not significantly different between in monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars). Data are mean ± SEM, n = 6, P>0.05 NT vs. HT. C, D, Representative micrographs of fMLP-induced polarization response of monocytes (C). Summary data of fMLP induced polarization response of monocytes from healthy control subjects (NT; open bars) and from patients with hypertension (HT, filled bars). Data are mean ± SEM, **p<0.01 compared to NT fMLP-stimulation (0 min); ##p<0.01 compared to HT fMLP-stimulation (0 min); P>0.05 NT vs. HT; each n = 12 (D).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3306381&req=5

pone-0032628-g004: Monocyte subtypes and fMLP receptors in normotensive and hypertensive patients.A, Peripheral blood monocytes subpopulations were analyzed by flow-cytometry. After labeling with anti-CD14 phycoerythrin (PE) conjugated and anti-CD16 FITC conjugated, monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars) were readily separated into three distinct subsets according to CD14 and CD16 positivity. Data are mean ± SEM, each n = 11, P>0.05 NT vs. HT. B, Expression of fMLP receptors using immunoblotting with specific antibodies. The data showed that fMLP receptors were not significantly different between in monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars). Data are mean ± SEM, n = 6, P>0.05 NT vs. HT. C, D, Representative micrographs of fMLP-induced polarization response of monocytes (C). Summary data of fMLP induced polarization response of monocytes from healthy control subjects (NT; open bars) and from patients with hypertension (HT, filled bars). Data are mean ± SEM, **p<0.01 compared to NT fMLP-stimulation (0 min); ##p<0.01 compared to HT fMLP-stimulation (0 min); P>0.05 NT vs. HT; each n = 12 (D).

Mentions: We evaluated CD14CD16 monocyte subset levels in normotensive control subjects and patients with essential hypertension (Figure 1D). We found that the percentages of CD14++CD16− and CD14+CD16+ subset levels were not significantly differed between normotensive control subjects and patients with essential hypertension (57±6% vs 50±5% for CD14++CD16−; and 12±2% vs 13±2% for CD14+CD16+; each n = 11, P>0.05; Figure 4A). In addition, the chemotaxis using an incubation time of 4 h was also used to test monocytes' migration. Also for a short incubation time of 4 hours we observed an increased fMLP-induced migration of monocytes from patients with essential hypertension compared to normotensive control subjects (159±12% vs 100±5%; each n = 8, P<0.01). We were interested whether differences of fMLP-induced migration in patients with essential hypertension and healthy subjects might be related to differences in the expression of the receptor for fMLP. Our data showed that fMLP receptors were not significantly different between patients with essential hypertension and normotensive control subjects (P>0.05, Figure 4B). These findings indicate that fMLP receptors may not be responsible for the observed differences of fMLP-induced monocytes migration between patients with essential hypertension and normotensive control subjects. On the other hand, microscopy showed that fMLP did not cause significant differences of the polarization response of monocytes from healthy control subjects and patients with hypertension (P>0.05, Figure 4C, 4D).


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)

Monocyte subtypes and fMLP receptors in normotensive and hypertensive patients.A, Peripheral blood monocytes subpopulations were analyzed by flow-cytometry. After labeling with anti-CD14 phycoerythrin (PE) conjugated and anti-CD16 FITC conjugated, monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars) were readily separated into three distinct subsets according to CD14 and CD16 positivity. Data are mean ± SEM, each n = 11, P>0.05 NT vs. HT. B, Expression of fMLP receptors using immunoblotting with specific antibodies. The data showed that fMLP receptors were not significantly different between in monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars). Data are mean ± SEM, n = 6, P>0.05 NT vs. HT. C, D, Representative micrographs of fMLP-induced polarization response of monocytes (C). Summary data of fMLP induced polarization response of monocytes from healthy control subjects (NT; open bars) and from patients with hypertension (HT, filled bars). Data are mean ± SEM, **p<0.01 compared to NT fMLP-stimulation (0 min); ##p<0.01 compared to HT fMLP-stimulation (0 min); P>0.05 NT vs. HT; each n = 12 (D).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3306381&req=5

pone-0032628-g004: Monocyte subtypes and fMLP receptors in normotensive and hypertensive patients.A, Peripheral blood monocytes subpopulations were analyzed by flow-cytometry. After labeling with anti-CD14 phycoerythrin (PE) conjugated and anti-CD16 FITC conjugated, monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars) were readily separated into three distinct subsets according to CD14 and CD16 positivity. Data are mean ± SEM, each n = 11, P>0.05 NT vs. HT. B, Expression of fMLP receptors using immunoblotting with specific antibodies. The data showed that fMLP receptors were not significantly different between in monocytes from patients with essential hypertension (HT, filled bars) and normotensive control subjects (NT; open bars). Data are mean ± SEM, n = 6, P>0.05 NT vs. HT. C, D, Representative micrographs of fMLP-induced polarization response of monocytes (C). Summary data of fMLP induced polarization response of monocytes from healthy control subjects (NT; open bars) and from patients with hypertension (HT, filled bars). Data are mean ± SEM, **p<0.01 compared to NT fMLP-stimulation (0 min); ##p<0.01 compared to HT fMLP-stimulation (0 min); P>0.05 NT vs. HT; each n = 12 (D).
Mentions: We evaluated CD14CD16 monocyte subset levels in normotensive control subjects and patients with essential hypertension (Figure 1D). We found that the percentages of CD14++CD16− and CD14+CD16+ subset levels were not significantly differed between normotensive control subjects and patients with essential hypertension (57±6% vs 50±5% for CD14++CD16−; and 12±2% vs 13±2% for CD14+CD16+; each n = 11, P>0.05; Figure 4A). In addition, the chemotaxis using an incubation time of 4 h was also used to test monocytes' migration. Also for a short incubation time of 4 hours we observed an increased fMLP-induced migration of monocytes from patients with essential hypertension compared to normotensive control subjects (159±12% vs 100±5%; each n = 8, P<0.01). We were interested whether differences of fMLP-induced migration in patients with essential hypertension and healthy subjects might be related to differences in the expression of the receptor for fMLP. Our data showed that fMLP receptors were not significantly different between patients with essential hypertension and normotensive control subjects (P>0.05, Figure 4B). These findings indicate that fMLP receptors may not be responsible for the observed differences of fMLP-induced monocytes migration between patients with essential hypertension and normotensive control subjects. On the other hand, microscopy showed that fMLP did not cause significant differences of the polarization response of monocytes from healthy control subjects and patients with hypertension (P>0.05, Figure 4C, 4D).

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