Ca2+ signaling in fowl aortic smooth muscle increases during maturation but is impaired in neointimal plaques.

1998 ◽  
Vol 201 (11) ◽  
pp. 1695-1705 ◽  
Author(s):  
Z L Qin ◽  
H Nishimura

Many bird species show the spontaneous development of high arterial pressure and vascular lesions in the aorta and other large arteries. In chickens, arterial pressure tends to increase with age/maturation (particularly in males), and subendothelial hyperplasia (neointima) in the abdominal aorta is often seen prior to sexual maturation. The mechanisms involved, however, are not known. Our aim, therefore, was to determine (1) whether cytosolic Ca2+ signaling (CCS) responses to vasoactive substances in fowl aortic smooth muscle differ among chickens at different maturation stages and (2) whether CCS responses to Ca2+ channel agonists in neointimal plaques differ from those in normal aortic smooth muscle. K+ increased CCS in a dose-dependent manner in isolated and superfused abdominal aortic smooth muscle tissue from chicks (5-9 weeks old), pullets (11-18 weeks old) and adult hens (20 weeks and older); CCS responses increased as chickens matured. The addition of Bay K 8644 (10(-6)mol l-1) to Ringer's solution containing 50 mmol l-1 K+ further increased CCS, and this response was reduced to half by nifedipine (10(-6)mol l-1). Norepinephrine did not alter CCS in chicks, whereas marked dose-dependent increases in CCS were noted in pullets. In contrast to the CCS responses to K+, the norepinephrine-induced CCS responses became smaller in adult hens. Isolated neointimal plaques showed only slight increases in CCS in response to 50 mmol l-1 K+ plus Bay K 8644, whereas clear responses were noted in aortic smooth muscle tissue underlying the plaques. These results suggest (1) that CCS responses to Ca2+ channel agonists increased with sexual maturation in fowl, but (2) that CCS responses to norepinephrine were low in mature hens and to K+ plus Bay K 8644 were low in spontaneously developed neointima, suggesting that phenotypic modulation of Ca2+ channel/norepinephrine receptors may have occurred during maturation/aging and in neointima.

2007 ◽  
Vol 292 (5) ◽  
pp. G1429-G1438 ◽  
Author(s):  
Takashi Ohama ◽  
Masatoshi Hori ◽  
Eiichi Momotani ◽  
Yoichiro Iwakura ◽  
Fengling Guo ◽  
...  

Motility disorders are frequently observed in intestinal inflammation. We previously reported that in vitro treatment of intestinal smooth muscle tissue with IL-1β decreases the expression of CPI-17, an endogenous inhibitory protein of smooth muscle serine/threonine protein phosphatase, thereby inhibiting contraction. The present study was performed to examine the pathophysiological importance of CPI-17 expression in the motility disorders by using an in vivo model of intestinal inflammation and to define the regulatory mechanism of CPI-17 expression by proinflammatory cytokines. After the induction of acute ileitis with 2,4,6,-trinitrobenzensulfonic acid, CPI-17 expression declined in a time-dependent manner. This decrease in CPI-17 expression was parallel with the reduction of cholinergic agonist-induced contraction of smooth muscle strips and sensitivity of permeabilized smooth muscle fibers to Ca2+. Among the various proinflammatory cytokines tested, TNF-α and IL-1β were observed to directly inhibit CPI-17 expression and contraction in cultured rat intestinal tissue. Moreover, both TNF-α and IL-1β inhibited CPI-17 expression and contraction of smooth muscle tissue isolated from wild-type and IL-1α/β double-knockout mice. However, IL-1β treatment failed to inhibit CPI-17 expression and contraction in TNF-α knockout mice. In β-escin-permeabilized ileal tissues, pretreatment with anti-phosphorylated CPI-17 antibody inhibited the carbachol-induced Ca2+ sensitization in the presence of GTP. These findings suggest that CPI-17 was downregulated during intestinal inflammation and that TNF-α plays a central role in this process. Downregulation of CPI-17 may play a role in motility impairments in inflammation.


2005 ◽  
Vol 103 (3) ◽  
pp. 548-555 ◽  
Author(s):  
Jingui Yu ◽  
Yasuyuki Tokinaga ◽  
Toshiyuki Kuriyama ◽  
Nobuhiko Uematsu ◽  
Kazuhiro Mizumoto ◽  
...  

Background The mechanisms of amino-amide local anesthetic agent-induced vasoconstriction remain unclear. The current study was designed to examine the roles of the protein kinase C (PKC), Rho kinase, and p44/42 mitogen-activated protein kinase (p44/42 MAPK) signaling pathways in calcium (Ca2+)-sensitization mechanisms in ropivacaine-induced vascular contraction. Methods Endothelium-denuded rat aortic rings, segments, and strips were prepared. The cumulative dose-response relations of contraction and intracellular Ca2+ concentration to ropivacaine were tested, using isometric force transducers and a fluorometer, respectively. The dose-dependent ropivacaine-induced phosphorylation of PKC and p44/42 MAPK and the membrane translocation of Rho kinase were also detected using Western blotting. Results Ropivacaine induced a dose-dependent biphasic contractile response and an increase in intracellular Ca2+ concentration of rat aortic rings, increasing at concentrations of 3 x 10 m to 3 x 10 m and decreasing from 10 m to 3 x 10 m, with a greater tension/intracellular Ca2+ concentration ratio than that induced with potassium chloride. The contraction was attenuated in a dose-dependent manner, by the PKC inhibitors bisindolylmaleimide I and calphostin C, the Rho-kinase inhibitor Y 27632, and the p44/42 MAPK inhibitor PD 098059. Ropivacaine also induced an increase in phosphorylation of PKC and p44/42 MAPK, and membrane translocation of Rho kinase in accordance with the contractile responses, which were also significantly inhibited by bisindolylmaleimide I and calphostin C, Y 27632, and PD 098059, correspondingly. Conclusion These findings demonstrated that PKC-, Rho kinase-, and p44/42 MAPK-mediated Ca2+-sensitization mechanisms are involved in the ropivacaine-induced biphasic contraction of rat aortic smooth muscle.


2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Nwe Nwe Soe ◽  
Mark Sowden ◽  
Bradford C Berk

Objective: Cyclophilin A (CyPA) is a Secreted OXidative stress-induced Factor (SOXF) secreted by cardiovascular cells in response to Angiotensin II (Ang II) and reactive oxygen species (ROS). Extracellular CyPA is a proinflammatory mediator that regulates vascular remodeling, abdominal aortic aneurysm, atherosclerosis and cardiac hypertrophy. Post-translational modification of CyPA by acetylation in response to ROS has been described. Moreover, acetylation of CyPA is important in HIV pathogenesis. The mechanism and regulation of CyPA acetylation as well as its role in cardiovascular diseases are currently unknown. We hypothesized that Ang II regulates oxidative stress-induced CyPA acetylation that alters its expression and/or secretion in vascular smooth muscle cells. Methods and results: Ang II (1μM) increased acetylation of CyPA (Acyl-CyPA) in a time dependent manner, with a peak at 8hr (3.5±0.6 fold increase) in rat aortic smooth muscle cells (RASMC) as shown by Western blot. Mouse aortic smooth muscle cells from mice lacking CyPA (CyPA-/-) and wild type controls (WT) confirmed that Ang II induced acetylation reactivity coincided exactly with CyPA reactivity. In AT1R and CyPA cotransfected HeLa cells, Ang II increased Acyl-CyPA in a time dependent manner consistent with that in RASMC. The ROS scavengers Tiron or N-acetylcysteine significantly inhibited Ang II induced Acyl-CyPA in a dose dependent manner in RASMC. Ang II-induced CyPA acetylation was enhanced by 2 hr pretreatment with histone deacetylase inhibitor trichostatin (TSA) or sirtinol in a dose dependent manner. Similarly, Ang II-induced CyPA secretion was enhanced by pretreatment with TSA (1μM) in a time dependent manner. Moreover, acetyltransferase p300 and PCAF (p300/CBP-asociated factor) inhibitor anacardic acid (6-nonadecyl salicylic acid) dramatically inhibited CyPA expression, and Ang II induced Acyl-CyPA in a dose dependent manner. Conclusion: These results suggest that Ang II-induced CyPA acetylation is oxidative stress dependent, and that acetylation enhanced CyPA expression and secretion. Detailed mechanistic studies of the regulation of CyPA acetylation will help to identify a future therapeutic target for CyPA regulated cardiovascular diseases.


2012 ◽  
Vol 303 (2) ◽  
pp. C170-C178 ◽  
Author(s):  
Won Sun Park ◽  
Da Hye Hong ◽  
Youn Kyoung Son ◽  
Min Hee Kim ◽  
Seung Hun Jeong ◽  
...  

We investigated the impairment of ATP-sensitive K+ (KATP) channels in aortic smooth muscle cells (ASMCs) from isoproterenol-induced hypertrophied rabbits. The amplitude of KATP channels induced by the KATP channel opener pinacidil (10 μM) was greater in ASMCs from control than from hypertrophied animals. In phenylephrine-preconstricted aortic rings, pinacidil induced relaxation in a dose-dependent manner. The dose-dependent curve was shifted to the right in the hypertrophied (EC50: 17.80 ± 3.28 μM) compared with the control model (EC50: 6.69 ± 2.40 μM). Although the level of Kir6.2 subtype expression did not differ between ASMCs from the control and hypertrophied models, those of the Kir6.1 and SUR2B subtypes were decreased in the hypertrophied model. Application of the calcitonin-gene related peptide (100 nM) and adenylyl cyclase activator forskolin (10 μM), which activates protein kinase A (PKA) and consequently KATP channels, induced a KATP current in both control and hypertrophied animals; however, the KATP current amplitude did not differ between the two groups. Furthermore, PKA expression was not altered between the control and hypertrophied animals. These results suggests that the decreased KATP current amplitude and KATP channel-induced vasorelaxation in the hypertrophied animals were attributable to the reduction in KATP channel expression but not to changes in the intracellular signaling mechanism that activates the KATP current.


1990 ◽  
Vol 123 (2) ◽  
pp. 218-224 ◽  
Author(s):  
Xiangbing Wang ◽  
Noriyuki Sato ◽  
Monte A. Greer ◽  
Susan E. Greer ◽  
Staci McAdams

Abstract. The mechanism by which 30% medium hyposmolarity induces PRL secretion by GH4C1 cells was compared with that induced by 100 nmol/l TRH or 30 mmol/l K+. Removing medium Ca2+, blocking Ca2+ channels with 50 μmol/l verapamil, or inhibiting calmodulin activation with 20 μmol/l trifluoperazine, 10 μmol/l chlorpromazine or 10 μmol/l pimozide almost completely blocked hyposmolarity-induced secretion. The smooth muscle relaxant, W-7, which is believed relatively specific in inhibiting the Ca2+-calmodulin interaction, depressed hyposmolarity-induced PRL secretion in a dose-dependent manner (r = −0.991, p<0.01 ). The above drugs also blocked or decreased high K+-induced secretion, but had much less effect on TRH-induced secretion. Secretion induced by TRH, hyposmolarity, or high K+ was optimal at pH 7.3-7.65 and was significantly depressed at pH 6.0 or 8.0, indicating that release of hormone induced by all 3 stimuli is due to an active cell process requiring a physiologic extracellular pH and is not produced by nonspecific cell toxicity. The data suggest hyposmolarity and high K+ may share some similarities in their mechanism of stimulating secretion, which is different from that of TRH.


2008 ◽  
Vol 26 (4) ◽  
pp. 307-314 ◽  
Author(s):  
Byung-Soo Kim ◽  
Anthony Atala ◽  
James J. Yoo

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