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ABSTRACT: Extracellular vesicles (EVs) carry signals within or at their limiting membranes, providing a mechanism by which cells can exchange more complex information than what was previously thought. In addition to mRNAs and microRNAs, there are DNA fragments in EVs. Solexa sequencing indicated the presence of at least 16434 genomic DNA (gDNA) fragments in the EVs from human plasma. Immunofluorescence study showed direct evidence that acridine orange-stained EV DNAs could be transferred into the cells and localize to and inside the nuclear membrane. However, whether the transferred EV DNAs are functional or not is not clear. We found that EV gDNAs could be homologously or heterologously transferred from donor cells to recipient cells, and increase gDNA-coding mRNA, protein expression, and function (e.g. AT1 receptor). An endogenous promoter of the AT1 receptor, NF-κB, could be recruited to the transferred DNAs in the nucleus, and increase the transcription of AT1 receptor in the recipient cells. Moreover, the transferred EV gDNAs have pathophysiological significance. BCR/ABL hybrid gene, involved in the pathogenesis of chronic myeloid leukemia, could be transferred from K562 EVs to HEK293 cells or neutrophils. Our present study shows that the gDNAs transferred from EVs to cells have physiological significance, not only to increase the gDNA-coding mRNA and protein levels, but also to influence function in recipient cells.
Journal of Molecular Cell Biology 04/2013;
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ABSTRACT: AIMS: Vascular inflammation is a key factor in the pathogenesis of diabetes-related vascular complications. Our previous study showed that (-)-epigallocatechin-3-gallate (EGCG) inhibits high glucose-induced vascular smooth muscle cells proliferation, thus may have beneficial effects in diabetes and its complications. However, the effect of EGCG on inflammation in diabetes is not known. In the present study, we investigated whether EGCG suppresses the vascular inflammation induced by high glucose in human umbilical vein endothelial cells (HUVECs). MAIN METHODS: The inhibitory effect of EGCG on high glucose-induced up-regulation of the expression of vascular cell adhesion molecule 1 (VCAM-1) was measured using enzyme-linked immunosorbent, RT-PCR, immunoblotting and cell adhesion assays. The effect of EGCG on high glucose-induced nuclear factor-kappa B (NF-κB) activation was investigated by immunoblotting, immunofluorescence and electrophoretic mobility shift assays. KEY FINDINGS: High glucose increased VCAM-1 expression and enhanced the adhesion of monocytes to HUVECs. Pretreatment with EGCG in a concentration-dependent manner (1.0-50 μM) significantly attenuated these effects. High glucose (25 mM)-mediated vascular inflammation was blocked by PKC pseudosubstrate (PKC inhibitor 19-31) or the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC). Stimulation with high glucose increased the NF-κB translocation from the cytoplasm to the nucleus, and increased IκB-α phosphorylation, decreased its expression, and in the presence of EGCG, the effect of high glucose on NF-κB and IκB-α were blocked. SIGNIFICANCE: EGCG suppresses high glucose-induced vascular inflammatory process via the inhibition of PKC and NF-κB activation in HUVECs, suggesting that EGCG may be a potential candidate for the treatment and prevention of diabetic vascular complications.
Life sciences 02/2013; · 2.56 Impact Factor
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ABSTRACT: The renin-angiotensin (Ang) system controls blood pressure, in part, by regulating renal tubular sodium transport. In the kidney, activation of the angiotensin II type 1 (AT(1)) receptor increases renal sodium reabsorption, whereas the angiotensin II type 2 (AT(2)) receptor produces the opposite effect. We hypothesized that the AT(2) receptor regulates AT(1) receptor expression and function in the kidney.
In immortalized renal proximal tubule (RPT) cells from Wistar-Kyoto rats, CGP42112, an AT(2) receptor agonist, decreased AT(1) receptor mRNA and protein expression (P < 0.05), as assessed by reverse transcriptase-polymerase chain reaction and immunoblotting. The inhibitory effect of the AT(2) receptor on AT(1) receptor expression was blocked by the AT(2) receptor antagonist, PD123319 (10 (-6)mol/l), the nitric oxide synthase inhibitor N(w)-nitro-L-arginine methyl ester (10(-4) mol/l), or the nitric oxide-dependent soluble guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo-[4,3-a] quinoxalin-1-one (10(-5) mol/l), indicating that both nitric oxide and cyclic guanosine monophosphate (cGMP) were involved in the signaling pathway. Furthermore, CGP42112 decreased Sp1 serine phosphorylation and reduced the binding of Sp1 to AT(1) receptor DNA. Stimulation with Ang II (10(-11) mol/l per 30 min) enhanced Na(+)-K(+)-ATPase activity in RPT cells, which was prevented by pretreatment with CGP42112 (10(-7) mol/l per 24 h) (P < 0.05). The above-mentioned results were confirmed in RPT cells from AT(2) receptor knockout mice; AT(1) receptor expression and Ang II-stimulated Na-K-ATPase activity were greater in these cells than in RPT cells from wild-type mice (P < 0.05). AT(1)/AT(2) receptors co-localized and co-immunoprecipitated in RPT cells; short-term CGP42112 (10 mol/l per 30 min) treatment increased AT(1)/AT(2) receptor co-immunoprecipitation (P < 0.05).
These results indicate that the renal AT(2) receptor, via nitric oxide/cGMP/Sp1 pathway, regulates AT(1 )receptor expression and function, which may be important in the regulation of sodium excretion and blood pressure.
Journal of hypertension 04/2012; 30(6):1176-84. · 4.02 Impact Factor
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Hefei Huang,
Hongmei Ren,
Caiyu Chen,
Xiaoyan Wang,
Jian Yang,
Yu Han, Duofen He,
Lin Zhou,
Laureano D Asico,
Pedro A Jose,
Chunyu Zeng
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ABSTRACT: Dopamine receptor, via D(1)-like and D(2)-like receptors, increases sodium excretion in kidney. We have reported positive interactions between D(3) and D(1) receptors in renal proximal tubule (RPT) cells. These reports, however do not preclude that there may be also interaction between D(3) and D(5) receptors, because of the lack of selective D(1) and D(5) receptor agonists or antagonists. We hypothesize that D(3) receptors can regulate D(5) receptors, and that D(3) receptor regulation of D(5) receptors in RPTs is impaired in spontaneously hypertensive rats (SHRs). It showed that a D(3) receptor agonist, PD128907, by the activation of protein kinase C activity, increased the expression of D(5) receptors in a concentration- and time-dependent manner in RPT cells from Wistar-Kyoto (WKY) rats. The stimulatory effect of the D(3) receptor on D(5) receptor expression was impaired in RPT cells from SHRs. The effect of D(3) receptor on D(5) receptor is functionally relevant; stimulation of D(5) receptor decreases Na(+)-K(+) adenosine triphosphatase (ATPase) activity in WKY cells. Pretreatment with D(3) receptor agonist for 24 h enhances the D(5) receptor expression and D(5) receptor-mediated inhibitory effect on Na(+)-K(+) ATPase activity in WKY cells, but decreases them in SHR cells. The effect of D(3) receptor on D(5) receptor expression and function was also confirmed in the D(5) receptor-transfected HEK293 cells. It indicates that activation of D(3) receptor increases D(5) receptor expression and function. Altered regulation of D(3) receptor on D(5) receptors may have a role in the pathogenesis of hypertension.
Hypertension Research 02/2012; 35(6):639-47. · 2.58 Impact Factor
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ABSTRACT: Proliferation of vascular smooth muscle cells (VSMCs) plays an important role in the development and progression of diabetes-related vascular complications. (-)-Epigallocatechin gallate (EGCG), the major catechin derived from green tea, is able to exert antidiabetes effects in animal models. However, it is not known whether or not EGCG inhibits VSMC proliferation induced by high glucose. This study tested the hypothesis that EGCG might have an inhibitory effect on VSMC proliferation induced by high glucose. VSMC proliferation was determined by [(3)H]-thymidine incorporation and uptake of 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT). Extracellular signal-regulated kinase (ERK) 1/2 phosphorylation was determined by immunoblotting, and ERK 1/2 activity was detected by measuring the ability to phosphorylate its substrate Elk-1. Glucose increased VSMC proliferation in a concentration-dependent manner, which was reduced in the presence of EGCG. VSMC proliferation mediated by high glucose (30 mM) was involved in protein kinase C (PKC) and ERK1/2 signalings, because its effect was blocked by PKC inhibitor (PKC inhibitor 19-31) and ERK1/2 inhibitor (PD98059). Pretreatment of VSMCs with EGCG significantly inhibited the stimulatory effect of high glucose on PKC and ERK1/2 activation, followed by attenuation of its downstream transcription factor Elk-1 phosphorylation. Taken together, these results suggest that EGCG could suppress VSMC proliferation induced by high glucose by inhibition of PKC and ERK1/2 signalings in VSMCs, which indicates that EGCG might be a possible medicine to reduce vascular complications in diabetes.
Journal of Agricultural and Food Chemistry 11/2011; 59(21):11483-90. · 2.82 Impact Factor
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ABSTRACT: The D(3) dopamine receptor is the major D(2)-like receptor that regulates sodium transport in the renal proximal tubule (RPT) and helps maintain blood pressure in the normal range. In Wistar-Kyoto (WKY) rats chronically fed high-salt diet, the intrarenal arterial infusion of a D(3) receptor agonist, PD128907, increased absolute and fractional sodium excretion. We have reported that Gα(12) and Gα(13), which participate in the signal transduction of the D(5) receptor, are expressed in RPTs. As the D(3) receptor is also expressed in RPTs, we hypothesized that it may also interact with Gα(12)/Gα(13) in RPTs from WKY rats. There were co-localization and co-immunoprecipitation of D(3) receptor and Gα(12)/Gα(13) in renal brush border membranes (BBMs) and RPT cells. The intrarenal infusion of PD128907 (1 μg kg(-1) min(-1)) that increased sodium excretion also increased the co-immunoprecipitations of D(3)/Gα(12) and D(3)/Gα(13) in renal BBMs; their co-immunoprecipitation was confirmed in RPT cells. As Gα(12) and Gα(13) increase sodium pump and transporter activity (for example, Na(+)-K(+)-ATPase, NHE3), an increased association of D(3) receptors with Gα(12)/Gα(13) receptors after D(3) receptor activation may be a mechanism to prevent Gα(12)/Gα(13)-mediated stimulation of sodium transport (and thus enhance natriuresis). We conclude that a D(3) receptor interaction with Gα(12)/Gα(13) that increases sodium excretion may have a role in the regulation of blood pressure.
Hypertension Research 06/2011; 34(9):1011-6. · 2.58 Impact Factor
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Hefei Huang,
Yu Han,
Xiaoyan Wang,
Caiyu Chen,
Changqing Yu, Duofen He,
Hongyong Wang,
Lin Zhou,
Laureano D Asico,
Pedro A Jose,
Chunyu Zeng
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ABSTRACT: Vascular smooth muscle cell (VSMC) proliferation is regulated by numerous hormones and humoral factors. Our previous study found that stimulation of D(1)-like dopamine receptors inhibited insulin receptor expression and function in VSMCs. We hypothesize that there is also an interaction between D(3) dopamine and insulin receptors, i.e., stimulation of the D(3) receptor inhibits insulin receptor expression and function.
Receptor expression was determined by immunoblotting, immunohistochemisty, and reverse transcriptase-PCR; VSMC proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-diphenyl-tetrazolium bromide (MTT) assay and cell number.
Insulin receptor protein is increased in the aorta of D(3) receptor deficient mice. Stimulation of the D(3) receptor inhibited insulin receptor mRNA and protein expression and insulin-mediated VSMC proliferation, and increased protein kinase A (PKA) activity, insulin receptor phosphorylation, and degradation in immortalized aortic VSMCs (A10 cells). These effects were blocked by a PKA inhibitor, indicating that the D(3) receptor-mediated decrease in insulin receptor expression was related to a decrease in transcription/post-transcription and increased degradation, involving PKA signaling.
D(3) receptor stimulation may be a target to reduce the adverse effect of insulin in hypertension by inhibition of insulin receptor expression and function in arterial VSMCs.
American Journal of Hypertension 03/2011; 24(6):654-60. · 3.18 Impact Factor
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ABSTRACT: Activation of renal D₃ receptor induces natriuresis and diuresis in Wistar-Kyoto (WKY) rats; in the presence of ETB receptor antagonist, the natriuretic effect of D₃ receptor in WKY rats is reduced. We hypothesize that ETB receptor activation may regulate D₃ receptor expression in renal proximal tubule (RPT) cells from WKY rats, which is impaired in RPT cells from spontaneously hypertensive rats (SHRs).
D₃ receptor expression was determined by immunoblotting; the D₃/ETB receptor linkage was checked by coimmunoprecipitation; Na(+)-K(+)-ATPase activity was determined as the rate of inorganic phosphate released in the presence or absence of ouabain.
In RPT cells from WKY rats, the ETB receptor agonist BQ3020 increased D₃ receptor protein. In contrast, in RPT cells from SHRs, BQ3020 did not increase D₃ receptor. There was coimmunoprecipitation between D₃ and ETB receptors in RPT cells from WKY and SHRs. Activation of ETB receptor increased D₃/ETB coimmunoprecipitation in RPT cells from WKY rats, but not from SHRs. The basal levels of D₃/ETB receptor coimmunoprecipitation were greater in RPT cells from WKY rats than in those from SHRs. Stimulation of D₃ receptor inhibited Na(+)-K(+)-ATPase activity, which was augmented by the pretreatment with the ETB receptor agonist BQ3020 in WKY RPT cells, but not in SHR RPT cells.
ETB receptors regulate and physically interact with D₃ receptors differently in WKY rats and SHRs. The impaired natriuretic effect in SHRs may be, in part, related to impaired ETB and D₃ receptor interactions.
Kidney and Blood Pressure Research 01/2011; 34(2):75-82. · 1.46 Impact Factor
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Yan Liu,
Jian Yang,
Hongmei Ren, Duofen He,
Annabelle Pascua,
M Ines Armando,
Chengming Yang,
Lin Zhou,
Robin A Felder,
Pedro A Jose,
Chunyu Zeng
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ABSTRACT: The kidney is important in the long-term regulation of blood pressure and sodium homeostasis. Stimulation of ETB receptors in the kidney increases sodium excretion, in part, by decreasing sodium transport in the medullary thick ascending limb of Henle and in collecting duct. However, the role of ETB receptor on Na+–K+ ATPase activity in renal proximal tubule (RPT) cells is not well defined. The purpose of this study is to test the hypothesis that ETB receptor inhibits Na+–K+ ATPase activity in rat RPT cells, and investigate the mechanism(s) by which such an action is produced. In RPT cells from Wistar–Kyoto rats, stimulation of ETB receptors by the ETB receptor agonist, BQ3020, decreased Na+–K+ ATPase activity, determined by ATP hydrolysis (control=0.38±0.02, BQ3020=0.26±0.03, BQ788=0.40±0.06, BQ3020+BQ788=0.37±0.04, n=5, P<0.01). The ETB receptor-mediated inhibition of Na+–K+ ATPase activity was dependent on an increase in intracellular calcium, because this effect was abrogated by a chelator of intracellular-free calcium (BAPTA-AM; 5 × 10−3 M 15 min−1), Ca2+ channel blocker (10−6 M 15 min−1 nicardipine) and PI3 kinase inhibitor (10−7 M per wortmannin). An inositol 1,4,5-trisphosphate (IP3) receptor blocker (2-aminoethyl diphenyl borate; 10−4 M 15 min−1) also blocked the inhibitory effect of the ETB receptor on Na+–K+ATPase activity (control=0.39±0.06, BQ3020=0.25±0.01, 2-APB=0.35±0.05, BQ3020+ 2-APB=0.35±0.06, n=4, P<0.01). The calcium channel agonist (BAY-K8644; 10−6 M 15 min−1) inhibited Na+–K+ ATPase activity, an effect that was blocked by a phosphatidylinositol-3 kinase inhibitor (10−7 M 15 min−1 wortmannin). In rat RPT cells, activation of the ETB receptor inhibits Na+–K+ ATPase activity by facilitating extracellular Ca2+ entry and Ca2+ release from endoplasmic reticulum.Keywords: ETB receptor, kidney, Na+–K+ ATPase activity, renal proximal tubule
Hypertension Research 08/2009; 32(10):846-852. · 2.58 Impact Factor
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Yan Liu,
Jian Yang,
Hongmei Ren, Duofen He,
Annabelle Pascua,
M Ines Armando,
Chengming Yang,
Lin Zhou,
Robin A Felder,
Pedro A Jose,
Chunyu Zeng
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ABSTRACT: The kidney is important in the long-term regulation of blood pressure and sodium homeostasis. Stimulation of ETB receptors in the kidney increases sodium excretion, in part, by decreasing sodium transport in the medullary thick ascending limb of Henle and in collecting duct. However, the role of ETB receptor on Na(+)-K(+) ATPase activity in renal proximal tubule (RPT) cells is not well defined. The purpose of this study is to test the hypothesis that ETB receptor inhibits Na(+)-K(+) ATPase activity in rat RPT cells, and investigate the mechanism(s) by which such an action is produced. In RPT cells from Wistar-Kyoto rats, stimulation of ETB receptors by the ETB receptor agonist, BQ3020, decreased Na(+)-K(+) ATPase activity, determined by ATP hydrolysis (control=0.38+/-0.02, BQ3020=0.26+/-0.03, BQ788=0.40+/-0.06, BQ3020+BQ788=0.37+/-0.04, n=5, P<0.01). The ETB receptor-mediated inhibition of Na(+)-K(+) ATPase activity was dependent on an increase in intracellular calcium, because this effect was abrogated by a chelator of intracellular-free calcium (BAPTA-AM; 5 x 10(-3) M 15 min(-1)), Ca(2+) channel blocker (10(-6) M 15 min(-1) nicardipine) and PI3 kinase inhibitor (10(-7) M per wortmannin). An inositol 1,4,5-trisphosphate (IP3) receptor blocker (2-aminoethyl diphenyl borate; 10(-4) M 15 min(-1)) also blocked the inhibitory effect of the ETB receptor on Na(+)-K(+)ATPase activity (control=0.39+/-0.06, BQ3020=0.25+/-0.01, 2-APB=0.35+/-0.05, BQ3020+ 2-APB=0.35+/-0.06, n=4, P<0.01). The calcium channel agonist (BAY-K8644; 10(-6) M 15 min(-1)) inhibited Na(+)-K(+) ATPase activity, an effect that was blocked by a phosphatidylinositol-3 kinase inhibitor (10(-7) M 15 min(-1) wortmannin). In rat RPT cells, activation of the ETB receptor inhibits Na(+)-K(+) ATPase activity by facilitating extracellular Ca(2+) entry and Ca(2+) release from endoplasmic reticulum.
Hypertension Research 08/2009; 32(10):846-52. · 2.58 Impact Factor
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Jian Yang,
Zhigang Cui, Duofen He,
Hongmei Ren,
Yu Han,
Changqing Yu,
Chunjiang Fu,
Zheng Wang,
Chengming Yang,
Xukai Wang,
Lin Zhou,
Laureano D Asico,
Van Anthony M Villar,
Ulrich Hopfer,
Mantian Mi,
Chunyu Zeng,
Pedro A Jose
[show abstract]
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ABSTRACT: Ion transport in the renal proximal tubule (RPT) is regulated by numerous hormones and humoral factors, including insulin and dopamine. Previous studies show an interaction between insulin and the D(1) receptor. Because both D(1) and D(5) receptors belong to the D(1)-like receptor subfamily, it is possible that an interaction between insulin and the D(5) dopamine receptor exists in RPT cells from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs).
D(5) receptor expression in immortalized RPT cells from WKY and SHRs was quantified by immunoblotting and D(5) receptor function by measuring Na(+)-K(+) ATPase activity.
Insulin increased the expression of the D(5) receptor. Stimulation with insulin (10(-7) mol/l) for 24 h increased D(5) receptor expression in RPT cells from WKY rats. This effect of insulin on D(5) receptor expression was aberrant in RPT cells from SHRs. The stimulatory effect of insulin on D(5) receptor expression in RPT cells from WKY rats was inhibited by a protein kinase C (PKC) inhibitor (PKC inhibitor peptide 19-31, 10(-6) mol/l) or a phosphatidylinositol 3 (PI3) kinase inhibitor (wortmannin, 10(-6) mol/l), indicating that both PKC and PI3 kinase were involved in the signaling pathway. Stimulation of the D(5) receptor heterologously expressed in HEK293 cells with fenoldopam (10(-7) mol/l/15 min) inhibited Na(+)-K(+) ATPase activity, whereas pretreatment with insulin (10(-7) mol/l/24 h) increased the D(5) receptor-mediated inhibition.
Insulin and D(5) receptors interact to regulate renal sodium transport; an aberrant interaction between insulin and D(5) receptor may participate in the pathogenesis of hypertension.
American Journal of Hypertension 05/2009; 22(7):770-6. · 3.18 Impact Factor
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Changqing Yu,
Zhiwei Yang,
Hongmei Ren,
Yie Zhang,
Yu Han, Duofen He,
Quansheng Lu,
Xiaoyan Wang,
Xukai Wang,
Chengming Yang,
Laureano D Asico,
Ulrich Hopfer,
Gilbert M Eisner,
Pedro A Jose,
Chunyu Zeng
[show abstract]
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ABSTRACT: The dopaminergic and endothelin systems, by regulating sodium transport in the renal proximal tubule (RPT), participate in the control of blood pressure. The D(3) and ETB receptors are expressed in RPTs, and D(3) receptor function in RPTs is impaired in spontaneously hypertensive rats (SHRs). Therefore, we tested the hypothesis that D(3) receptors can regulate ETB receptors, and that D(3) receptor regulation of ETB receptors in RPTs is impaired in SHRs.
ETB receptor expression in RPT cells was measured by immunoblotting and reverse transcriptase-PCR and ETB receptor function by measuring Na(+)-K(+) ATPase activity. D(3)/ETB receptor interaction was studied by co-immunoprecipitation.
In Wistar-Kyoto (WKY) RPT cells, the D(3) receptor agonist, PD128907, increased ETB receptor protein expression, effects that were blocked by removal of calcium in the culture medium. The stimulatory effect of D(3) on ETB receptor mRNA and protein expression was also blocked by nicardipine. In contrast, in SHR RPT cells, PD128907 decreased ETB receptor expression. Basal D(3)/ETB receptor co-immunoprecipitation was three times greater in WKY than in SHRs. The absolute amount of D(3)/ETB receptor co-immunoprecipitation induced by a D(3) receptor agonist was also greater in WKY than in SHRs. Stimulation of ETB receptors decreased Na(+)-K(+) ATPase activity in WKY but not in SHR cells. Pretreatment with PD128907 augmented the inhibitory effect of BQ3020 on Na(+)-K(+) ATPase activity in WKY but not in SHR cells.
D(3) receptors regulate ETB receptors by physical receptor interaction and govern receptor expression and function. D(3) receptor regulation of ETB receptors is aberrant in RPT cells from SHRs.
American Journal of Hypertension 05/2009; 22(8):877-83. · 3.18 Impact Factor
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Changqing Yu,
Zhiwei Yang,
Hongmei Ren,
Yie Zhang,
Yu Han, Duofen He,
Quansheng Lu,
Xiaoyan Wang,
Xukai Wang,
Chengming Yang,
Laureano D. Asico,
Ulrich Hopfer,
Gilbert M. Eisner,
Pedro A. Jose,
Chunyu Zeng
[show abstract]
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ABSTRACT: Background The dopaminergic and endothelin systems, by regulating sodium transport in the renal proximal tubule (RPT), participate in the control of blood pressure. The D3 and ETB receptors are expressed in RPTs, and D3 receptor function in RPTs is impaired in spontaneously hypertensive rats (SHRs). Therefore, we tested the hypothesis that D3 receptors can regulate ETB receptors, and that D3 receptor regulation of ETB receptors in RPTs is impaired in SHRs.
American Journal of Hypertension 04/2009; 22(8):877-883. · 3.18 Impact Factor
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Jian Yang,
Zhigang Cui, Duofen He,
Hongmei Ren,
Yu Han,
Changqing Yu,
Chunjiang Fu,
Zheng Wang,
Chengming Yang,
Xukai Wang,
Lin Zhou,
Laureano D. Asico,
Van Anthony M. Villar,
Ulrich Hopfer,
Mantian Mi,
Chunyu Zeng,
Pedro A. Jose
[show abstract]
[hide abstract]
ABSTRACT: Background Ion transport in the renal proximal tubule (RPT) is regulated by numerous hormones and humoral factors, including insulin and dopamine. Previous studies show an interaction between insulin and the D1 receptor. Because both D1 and D5 receptors belong to the D1-like receptor subfamily, it is possible that an interaction between insulin and the D5 dopamine receptor exists in RPT cells from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs).
American Journal of Hypertension 04/2009; 22(7):770-776. · 3.18 Impact Factor
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Chunyu Zeng,
Yu Han,
Hefei Huang,
Changqing Yu,
Hongmei Ren,
Weibin Shi, Duofen He,
Lan Huang,
Chengming Yang,
Xukai Wang,
Lin Zhou,
Pedro A Jose
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ABSTRACT: Vascular smooth muscle cell (VSMC) proliferation is central to the development of vascular diseases, including hypertension, which is regulated by numerous hormones and humoral factors. Our previous study showed that the stimulatory effect of norepinephrine on VSMC proliferation is inhibited by D1-like receptors and the D3 dopamine receptor, a member of the D2-like receptor family. Insulin is a proliferative hormone but it is not known if there is any interaction between insulin and D1-like receptors. We hypothesized that Dl-like receptors may have an inhibitory effect on the insulin-induced VSMC proliferation; aberrant insulin and Dl-like receptor functions could be involved in the pathogenesis of essential hypertension.
VSMC proliferation was determined by [H]-thymidine incorporation; insulin receptor mRNA and protein expressions were determined by RT-PCR, immunoblotting, and immunohistochemistry.
Insulin increased VSMC proliferation in immortalized aortic A10 cells, determined by [H]-thymidine incorporation. Although the D1-like receptor, by itself, had no effect on VSMC proliferation, stimulation with fenoldopam, a D1-like receptor agonist, inhibited the stimulatory effect of insulin. The inhibitory effect of fenoldopam on insulin-mediated VSMC proliferation was receptor specific, because its effect could be blocked by SCH23390, a D1-like receptor antagonist. Fenoldopam also inhibited insulin receptor mRNA and protein expression, which was time dependent and concentration dependent. A PKC or MAP kinase inhibitor blocked the inhibitory effect of fenoldopam on insulin receptor expression, indicating that PKC and MAP kinase were involved in the signaling pathway.
The inhibitory effect of D1-like receptors on insulin-mediated VSMC proliferation may play an important role in the regulation of blood pressure.
Journal of hypertension 04/2009; 27(5):1033-41. · 4.02 Impact Factor
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Zhen Li,
Changqing Yu,
Yu Han,
Hongmei Ren,
Weibin Shi,
Chunjiang Fu, Duofen He,
Lan Huang,
Chengming Yang,
Xukai Wang,
Lin Zhou,
Laureano D Asico,
Chunyu Zeng,
Pedro A Jose
[show abstract]
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ABSTRACT: The sympathetic nervous system plays an important role in the regulation of blood pressure. There is increasing evidence for positive and negative interactions between dopamine and adrenergic receptors; the activation of the alpha-adrenergic receptor induces vasoconstriction, whereas the activation of dopamine receptor induces vasorelaxation. We hypothesize that the D1-like receptor and/or D3 receptor also inhibit alpha1-adrenergic receptor-mediated proliferation in vascular smooth muscle cells (VSMCs). In this study, VSMC proliferation was determined by measuring [3H]thymidine incorporation, cell number, and uptake of 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT). Norepinephrine increased VSMC number and MTT uptake, as well as [3H]thymidine incorporation via the alpha1-adrenergic receptor in aortic VSMCs from Sprague-Dawley rats. The proliferative effects of norepinephrine were attenuated by the activation of D1-like receptors or D3 receptors, although a D1-like receptor agonist, fenoldopam, and a D3 receptor agonist, PD-128907, by themselves, at low concentrations, had no effect on VSMC proliferation. Simultaneous stimulation of both D1-like and D3 receptors had an additive inhibitory effect. The inhibitory effect of D3 receptor was via protein kinase A, whereas the D1-like receptor effect was via protein kinase C-zeta. The interaction between alpha1-adrenergic and dopamine receptors, especially D1-like and D3 receptors in VSMCs, could be involved in the pathogenesis of hypertension.
AJP Heart and Circulatory Physiology 07/2008; 294(6):H2761-8. · 3.71 Impact Factor
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Chunyu Zeng,
Laureano D Asico,
Changqing Yu,
Van Anthony M Villar,
Weibin Shi,
Yingjin Luo,
Zheng Wang, Duofen He,
Yan Liu,
Lan Huang,
Chengming Yang,
Xukai Wang,
Ulrich Hopfer,
Gilbert M Eisner,
Pedro A Jose
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[hide abstract]
ABSTRACT: Dopaminergic and endothelin systems participate in the control blood pressure by regulating sodium transport in the renal proximal tubule. Disruption of either the endothelin B receptor (ETB) or D(3) dopamine receptor gene in mice produces hypertension. To examine whether these two receptors interact we studied the Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats by selectively infusing reagents into the right kidney of anesthetized rats. The D(3) receptor agonist (PD128907) caused natriuresis in WKY rats which was partially blocked by the ETB receptor antagonist. In contrast, PD128907 blunted sodium excretion in the SHRs. We found using laser confocal microscopy that the ETB receptor was mainly located in the cell membrane in control WKY cells. Treatment with the D(3) receptor antagonist caused its internalization into intracellular compartments that contained the D(3) receptors. Combined use of D(3) and ETB antagonists failed to internalize ETB receptors in cells from WKY rats. In contrast in SHR cells, ETB receptors were found mainly in internal compartments under basal condition and thus were likely prevented from interacting with the agonist-stimulated, membrane-bound D(3) receptors. Our studies suggest that D(3) receptors physically interact with proximal tubule ETB receptors and that the blunted natriuretic effect of dopamine in SHRs may be explained, in part, by abnormal D(3)/ETB receptor interactions.
Kidney International 07/2008; 74(6):750-9. · 6.61 Impact Factor
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Chunyu Zeng,
Yan Liu,
Zheng Wang, Duofen He,
Lan Huang,
Peiying Yu,
Shaopeng Zheng,
John E Jones,
Laureano D Asico,
Ulrich Hopfer,
Gilbert M Eisner,
Robin A Felder,
Pedro A Jose
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ABSTRACT: The dopaminergic and renin angiotensin systems interact to regulate blood pressure. Disruption of the D(3) dopamine receptor gene in mice produces renin-dependent hypertension. In rats, D(2)-like receptors reduce angiotensin II binding sites in renal proximal tubules (RPTs). Because the major D(2)-like receptor in RPTs is the D(3) receptor, we examined whether D(3) receptors regulate angiotensin II type 1 (AT(1)) receptors in rat RPT cells. The effect of D(3) receptors on AT(1) receptors was studied in vitro and in vivo. The D(3) receptor agonist PD128907 decreased AT(1) receptor protein and mRNA in WKY RPT cells and increased it in SHR cells. PD128907 increased D(3) receptors in WKY cells but had no effect in SHR cells. D(3)/AT(1) receptors colocalized in RPT cells; D(3) receptor stimulation decreased the percent amount of D(3) receptors that coimmunoprecipitated with AT(1) receptors to a greater extent in WKY than in SHR cells. However, D(3) receptor stimulation did not change the percent amount of AT(1) receptors that coimmunoprecipitated with D(3) receptors in WKY cells and markedly decreased the coimmunoprecipitation in SHR cells. The D(3) receptor also regulated the AT(1) receptor in vivo because AT(1) receptor expression was increased in kidneys of D(3) receptor-null mice compared with wild type littermates. D(3) receptors may regulate AT(1) receptor function by direct interaction with and regulation of AT(1) receptor expression. One mechanism of hypertension may be related to increased renal expression of AT(1) receptors due decreased D(3) receptor regulation.
Circulation Research 10/2006; 99(5):494-500. · 9.49 Impact Factor
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Hefei Huang,
Yu Han,
Xiaoyan Wang,
Caiyu Chen,
Changqing Yu, Duofen He,
Hongyong Wang,
Lin Zhou,
Laureano D Asico,
Pedro A Jose,
Chunyu Zeng
