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At least 379 records · Page 21Linked to original sources
PubMed2002

Uptake of (3)H-1-methyl-4-phenylpyridinium ((3)H-MPP(+)) by human intestinal Caco-2 cells is regulated by phosphorylation/dephosphorylation mechanisms.

Several transmembrane transporters of organic compounds are regulated by phosphorylation/dephosphorylation mechanisms. The aim of this study was to investigate the possible regulation of the intestinal uptake of organic cations by these mechanisms. The intestinal apical uptake of 1-methyl-4-phenylpyridinium (MPP(+)) was studied by incubating Caco-2 cells at 37 degrees for 5 min with 200 nM (3)H-MPP(+). Uptake of (3)H-MPP(+) by Caco-2 cells was not affected by activators of protein kinase G, and was not affected or slightly reduced (by 15-20%) by activators of protein kinase A or protein kinase C. Uptake of (3)H-MPP(+) by Caco-2 cells was reduced in a concentration-dependent manner by non-selective phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine (IBMX), caffeine, teophylline). The IC(50) of IBMX was found to be 119 microM (102-138; n=9). Uptake of (3)H-MPP(+) by Caco-2 cells was not affected by inhibition of protein tyrosine kinase, but it was concentration-dependently reduced in the presence of inhibitors of mitogen-activated protein kinase. Uptake of (3)H-MPP(+) by Caco-2 cells was strongly reduced by Ca(2+)/calmodulin-mediated pathway inhibitors, but it was not dependent on extracellular Ca(2+). Our results suggest that the intestinal apical uptake of MPP(+) is regulated by phosphorylation/dephosphorylation mechanisms, being most probably active in the dephosphorylated state. Moreover, uptake of (3)H-MPP(+) by Caco-2 cells and by the extraneuronal monoamine transporter (EMT) are regulated in a very similar manner, suggesting an important participation of EMT in the intestinal uptake of this compound.

1-Methyl-4-phenylpyridinium↗
PubMed2002

The alkaloid sanguinarine is effective against multidrug resistance in human cervical cells via bimodal cell death.

Sanguinarine, a benzophenanthrine alkaloid, is potentially antineoplastic through induction of cell death pathways. The development of multidrug resistance (MDR) is a major obstacle to the success of chemotherapeutic agents. The aim of this study was to investigate whether sanguinarine is effective against uterine cervical MDR and, if so, by which mechanism. The effects of treatment with sanguinarine on human papillomavirus (HPV) type 16-immortalized endocervical cells and their MDR counterpart cells were compared. Trypan blue exclusion assays and clonogenic survival assays demonstrated that MDR human cervical cells are as sensitive as their drug-sensitive parental cells to death induced by sanguinarine. Upon treatment of both types of cells with sanguinarine, two distinct concentration-dependent modes of cell death were observed. Treatment with 2.12 or 4.24 microM sanguinarine induced death in most cells that was characterized as apoptosis using the criteria of cell surface blebbing, as determined by light and scanning electron microscopy, and proteolytic activation of caspase-3 and cleavage of the caspase-3 substrate poly(ADP-ribose) polymerase (PARP), as detected by Western blot analysis. However, 8.48 and 16.96 microM sanguinarine caused a second mode of cell death, oncosis, distinguished by cell surface blistering, and neither caspase-3 activation nor PARP cleavage. This study provides the first evidence that sanguinarine is effective against MDR in cervical cells via bimodal cell death, which displays alternative mechanisms involving different morphologies and caspase-3 activation status.

Alkaloids↗
PubMed1998

Effect of taurine on chelerythrine inhibition of calcium uptake and ATPase activity in the rat retina.

Taurine potentiates calcium uptake in whole retinal homogenates as well as in rod outer segments and mitochondrial fractions. The aim of this study was to correlate taurine potentiation of calcium uptake with its effects on other cellular processes through the use of chelerythrine (CHT), a modulator of protein kinase C (PKC), ATPase activity, and, as recently shown, of retinal protein phosphorylation. CHT inhibited calcium uptake only when ATP was present, and inhibition increased significantly in conditions of ATP excess. Taurine potentiated ATP-dependent calcium uptake but decreased the potency of ATP to induce uptake activity. CHT inhibition of calcium uptake exhibited similar potencies in the presence and absence of taurine, and this inhibition seemed to be independent of PKC inhibition. Because of the ATP-dependence of the observed effect, total ATPase activity was studied using similar treatments. In the absence of taurine, CHT inhibited ATPase activity with the same potency (IC50 approximately 59.3 microM) as with calcium uptake inhibition (IC50 approximately 87.9 microM), presenting a possible mechanism of action of CHT. In the presence of taurine, no such correlation was observed, suggesting an ATPase-independent mechanism of action. In fact, taurine did not potentiate ATPase activity, but rather it decreased the potency of CHT inhibition of ATPase, effects incongruent with the effects of taurine on calcium uptake and on CHT inhibition of calcium uptake. Enzyme kinetic experiments provided more supporting data. Taurine was found to cause an increase in the affinity of the ATP substrate for the ATPase enzyme, contradicting the aforementioned effect of taurine to decrease the potency of ATP to induce calcium uptake. Thus, taurine seems to increase calcium uptake through a hitherto unreported mechanism distinct from its modulation of ATPase activity.

Adenosine Triphosphatases↗
PubMed1999

Regulation of cytochrome P4501B1 (CYP1B1) in mouse embryo fibroblast (C3H10T1/2) cells by protein kinase C (PKC).

The effects of co-treatment of C3H10T1/2 (10T1/2) cells with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 12-O-tetradecanoylphorbol-13-acetate (TPA) on the expression of the novel cytochrome P4501B1 (CYP1B1) were investigated. As monitored by CYP1B1-catalyzed 7,12-dimethylbenzanthracene (DMBA) metabolism, TPA suppressed basal and TCDD-induced DMBA metabolism in a concentration-dependent manner, with a maximum inhibitory concentration of 100 nM. The suppression of CYP1B1 catalytic activity occurred at two time points during which protein kinase C (PKC) was activated and down-regulated in these cells as judged by analyses of cellular PKC content and PKC-inhibitor (chelerythrine chloride)-influenced suppression of CYP1B1 catalytic activity. Experiments in which TCDD and benzanthracene (BA)-induced DMBA metabolism were monitored in PKCbeta1-overexpressing 10T1/2 cells revealed that the suppression of CYP1B1 activity is a consequence of cellular PKC elevation. This suppression phenomenon could be accounted for by PKC-mediated suppression of TCDD-induced CYP1B1 mRNA and apoprotein and of nuclear translocation of the Ah-receptor. In contrast, the mitogen-activated protein kinase (MAPK) proteins ERKs 1 and 2 were stimulated by TCDD under conditions in which PKC was activated. Collectively, our results suggest that PKC participates in the regulation of CYP1B1 in 10T1/2 cells, positively by directly suppressing the Ah-receptor signaling pathway, followed by an indirect or negative activation of the MAPK signaling pathway.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed2002

Factors contributing to neurotrophin-independent survival of adult sensory neurons.

Dorsal root ganglion (DRG) sensory neurons become less dependent upon neurotrophins for their survival as they mature. DRG neurons from young adult rats were dissociated and cultured in vitro in serum-free defined medium. We show that adult DRG sensory neurons are able to survive for at least 2 weeks in culture in the absence of nerve growth factor (NGF). We then investigated potential mechanisms contributing to this apparent neurotrophin-independent survival in these neurons through the use of inhibitors of cellular signaling pathways. The phosphoinositide kinase-3 (PI 3-K) inhibitor LY294002, and a protein kinase C (PKC) inhibitor, chelerythrine resulted in significant decreases in neuronal survival. Neither the mitogen activated protein kinase kinase (MEK) inhibitor U0126 nor two other PKC inhibitors (bisindolylmaleimide and rottlerin) had any significant effect on survival. Our results point to the importance of PI 3-K and PKC signaling in the neurotrophin-independent survival of adult DRG neurons.

Age Factors↗
PubMed2002

Involvement of 5-HT7 receptors in serotonergic effects on spike afterpotentials in presumed jaw-closing motoneurons of rats.

Intracellular recordings were obtained from rat presumed jaw-closing motoneurons in slice preparations to investigate the involvement of the serotonin(7) (5-HT(7)) receptors in serotonergic inhibition of the postspike medium-duration afterhyperpolarization (mAHP) and enhancement of the afterdepolarization (ADP). 5-HT-induced suppression of the mAHP and enhancement of the ADP were mimicked by application of the 5-HT(1A/7) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and antagonized by the 5-HT(2/6/7) receptor antagonist clozapine, whereas the 5-HT(2) receptor agonist alpha-methyl-5-hydroxytryptamine (alpha-methyl-5-HT) did not affect the mAHP and ADP. 8-OH-DPAT-induced attenuation of the mAHP and enhancement of the ADP were also antagonized by clozapine and another 5-HT(2/6/7) receptor antagonist ritanserin, whereas the 5-HT(1A) receptor antagonist pindolol failed to block the 8-OH-DPAT-induced effects on the mAHP and ADP. 8-OH-DPAT-induced suppression of the mAHP and enhancement of the ADP were also antagonized by a protein kinase A (PKA) inhibitor H89, whereas 8-OH-DPAT could inhibit the mAHP and enhance the ADP in the presence of a protein kinase C (PKC) inhibitor chelerythrine. The 8-OH-DPAT-induced suppression of the mAHP was enhanced under raised [Ca(2+)](o) and this enhancement was reduced by chelerythrine. It is suggested that the 5-HT(7) receptors are involved in 5-HT-induced attenuation of the mAHP and enhancement of the ADP through activation of PKA, and the attenuation of mAHP through the 5-HT(7) receptors is enhanced under raised [Ca(2+)](o) by PKC activation.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed2003

D-2-amino-5-phosphonopentanoic acid inhibits intrathecal pertussis toxin-induced thermal hyperalgesia and protein kinase Cgamma up-regulation.

The aim of the present study was to examine the effect of intrathecal (i.t.) injection of pertussis toxin (PTX) on the nociceptive threshold and protein kinase C (PKC) expression in the rat spinal cord. The role of N-methyl-D-aspartic acid (NMDA) receptors in these changes was also examined. Male Wistar rats were implanted with two i.t. catheters, one of which was connected to a mini-osmotic pump and used to infuse saline or D-2-amino-5-phosphonopentanoic acid (D-AP5) (2 microg/h) starting on day 3 after i.t. catheter insertion. Two days later, a single injection of saline or PTX (2 microg) was given via the other catheter, followed by a flush with 10 microl of saline. On day 4 after PTX or saline injection, the thermal paw withdrawal latency was measured, then the rats were sacrificed by decapitation, and the dorsal part of the lumbosacral spinal segments was removed for PKC Western blotting assays. In PTX-treated rats, thermal hyperalgesia was observed, and the PKCgamma content of both the synaptosomal membrane and cytosolic fractions was significantly increased. The levels of alpha-, betaI-, or betaII-PKC isozymes in these fractions were unaffected by PTX treatment. Infusion of the NMDA antagonist, D-AP5, prevented both the thermal hyperalgesia and the increase in PKCgamma isoform expression in PTX-treated rats, and had no effect on these values in nai;ve rats. Intrathecal injection of the PKC inhibitor, chelerythrine (10 microg), significantly inhibited the thermal hyperalgesia observed in PTX-treated rats. These results show that i.t. injection of PTX induced thermal hyperalgesia accompanied by a selective increase in PKCgamma expression in both the synaptosomal membrane and cytosolic fractions of the dorsal horn of the rat lumbar spinal cord, and both effects were inhibited by the NMDA receptor antagonist, D-AP5.

2-Amino-5-phosphonovalerate↗
PubMed2003

Protein kinase C activation generates superoxide and contributes to impairment of cerebrovasodilation induced by G protein activation after brain injury.

Previous studies have observed that activation of protein kinase C (PKC) contributes to generation of superoxide anion (O(-)(2)) after fluid percussion brain injury (FPI). This study was designed to characterize the effects of FPI on the vascular activity of two activators of a pertussis toxin sensitive G protein, mastoparan and mastoparan-7, and the role of PKC dependent O(-)(2) generation in such effects in newborn pigs equipped with a closed cranial window. Mastoparan (10(-8), 10(-6) M) elicited pial artery dilation that was blunted by FPI and partially restored by the PKC inhibitor chelerythrine (10(-7) M) or the O(-)(2) free radical scavengers polyethylene glycol superoxide dismutase and catalase (SODCAT) (9+/-1 and 16+/-1, sham control; 3+/-1 and 5+/-1, FPI; and 7+/-1 and 11+/-1%, FPI SODCAT pretreated). Similar results were observed for mastoparan-7 but the inactive analogue mastoparan-17 had no effect on pial artery diameter. Exposure of the cerebral cortex to a xanthine oxidase O(-)(2) generating system blunted mastoparan induced pial artery dilation similar to FPI (10+/-1 and 17+/-1 vs. 2+/-1 and 3+/-1%). Pertussis toxin (1 microg/ml) exposure blocked mastoparan and mastoparan-7 vasodilation. These data show that pertussis toxin sensitive G protein activation elicits cerebrovasodilation that is blunted following FPI in a PKC dependent manner. These data also show that O(-)(2) generation similarly blunts G protein mediated cerebrovasodilation. These data suggest that PKC dependent O(-)(2) generation contributes to impaired G protein mediated cerebrovasodilation after FPI.

Alkaloids↗
PubMed2003

The effects of protein kinase C activity on synaptic transmission in two areas of rat hippocampus.

The effects of three protein kinase C (PKC) agonists (phorbol ester, ingenol and indolactam-V) and two PKC antagonists (D-erythro-sphingosine and chelerythrine) on input-output (I-O) relations in the Schaffer collateral pathway to CA1 (SC-CA1) and mossy fiber pathway to CA3 (MF-CA3) were determined in rat hippocampus brain slices. In the SC-CA1 pathway, phorbol esters and indolactam-V had only small effects on field excitatory post-synaptic potentials (fEPSP) in slices from 60-day animals, although ingenol, an activator of novel PKC isozymes, caused a significant decrease of the field excitatory post-synaptic potentials amplitude in 60-day animals, but not in 30-day animals. In contrast, in the MF-CA3 pathway, PKC agonists induced a significant increase in the field excitatory post-synaptic potentials. PKC antagonists depressed the field excitatory post-synaptic potentials in the SC-CA1 pathway, but had no significant effect in the MF-CA3 pathway. In the MF-CA3 pathway, paired-pulse facilitation was abolished by PKC agonists and unaffected by antagonists. In SC-CA1, it was depressed by agonists to levels below control, whereas it was significantly increased by chelerythine. We conclude that PKC plays important but different roles in both regions. In the SC-CA1 pathway, PKC is almost maximally active under control circumstances, and PKC antagonists significantly reduce synaptic responses. In contrast, in the MF-CA3 pathway, there is no apparent activation under resting circumstances, but significant potentiation of synaptic transmission is induced when PKC is activated. There are developmental changes in the pattern of PKC isozymes, and both pre- and post-synaptic actions are important.

Alkaloids↗
PubMed1997

Protein kinases A and C are involved in the mechanisms underlying consolidation of cocaine place conditioning.

Using a balanced conditioned place preference (CPP) paradigm, we studied the role of protein kinases A (PKA) and C (PKC) on the acquisition, consolidation and expression of cocaine place conditioning. H7, a non-selective inhibitor of protein kinases, was administered intracerebroventricularly at 1 and 10 micrograms/10 microliters. The higher dose significantly reduced the time spent by rats in the cocaine compartment when given immediately after each conditioning session (consolidation), whereas it had no effect when administered before cocaine during the training phase (acquisition) or before testing for place preference in the absence of cocaine (expression). The same effect was found on administering immediately after each training session 3 micrograms/10 microliters chelerythrine, a selective PKC inhibitor, or 10 micrograms/10 microliters H89, a selective PKA inhibitor, suggesting that both kinases contribute to the consolidation of stimulus-reward association which determines rats' behavior in the cocaine CPP. Changes in the activity of PKA and PKC may thus be part of the cascade of events that contribute to enhancing synaptic responses in the consolidation phase of cocaine CPP and determine rats' behavior associated with the memory of the rewarding effect of cocaine during cocaine CPP expression. These findings may have implications for the study of cocaine 'craving' and relapse.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1998

Chelerythrine, a specific PKC inhibitor, blocks acquisition but not consolidation and retrieval of conditioned taste aversion in rat.

Association of the short-term memory of the gustatory conditioned stimulus (CS) with visceral malaise (unconditioned stimulus, US) in conditioned taste aversion (CTA) paradigm takes place in the parabrachial nuclei (PBN) of brainstem. In order to ascertain the role of protein-kinase C (PKC) during different phases of CTA acquisition and retrieval, four experimental series were carried out. In Experiment 1, 1 microl of 10 mM of PKC inhibitor chelerythrine prevented CTA acquisition when applied into PBN in the CS-US interval. In Experiment 2, the necessity of PKC activity in different phases of CTA acquisition was tested by prolonging the time interval between PBN administration of chelerythrine and i.p. LiCl. CTA acquisition was prevented when chelerythrine-induced blockade of PKC coincided with GSTM persistence but not with CTA consolidation. In Experiment 3, the interval between saccharin drinking and LiCl injection was prolonged to 120 min. Again, chelerythrine blockade of PKC activity prevented CTA formation when it interfered with GSTM persistence. In Experiment 4, the possibility that PKC activity is necessary also for CTA retrieval was tested by chelerythrine application into PBN 5 min before retrieval testing. In this case, the chelerythrine-induced PKC blockade did not impair CTA retrieval. It is concluded that PKC is important for GSTM formation and persistence but not for CTA consolidation or retrieval.

Alkaloids↗
PubMed1999

Taurine uptake activity in the rat retina: protein kinase C-independent inhibition by chelerythrine.

Taurine, a regulatory amino acid of various biochemical processes in the retina, requires an efficient uptake system to maintain the high physiological concentration of taurine in the retina. Taurine uptake was characterized in both whole retinal preparations and in isolated rod outer segments (ROS) in terms of uptake kinetics and possible protein kinase C (PKC)-dependent regulation. Two uptake systems, a high- and a low-affinity system, were found in whole retinal preparations while only the high-affinity system was found in the isolated ROS. All the uptake systems characterized were inhibited by guanidinoethane sulfonate (GES), a well-known competitive inhibitor of taurine uptake. Stimulation and inhibition of PKC activity with phorbol myristate acetate and with staurosporine, respectively, produced no significant effect on taurine uptake. On the other hand, chelerythrine (CHT), a documented potent PKC inhibitor, was found to cause significant inhibition of the two taurine uptake systems, presumably through a PKC-independent mechanism. The data demonstrate that CHT may be a useful tool in studying taurine uptake in the retina and specifically in the ROS.

Alkaloids↗
PubMed1999

Axonal transport is inhibited by a protein kinase C inhibitor in cultured isolated mouse dorsal root ganglion cells.

We investigated roles of protein kinase C (PKC) and Ca2+/calmodulin-dependent protein II (CAM II) kinase activities in the maintenance of axonal transport in cultured isolated mouse dorsal root ganglion (DRG) cells. Video-enhanced microscopic recordings revealed that the PKC inhibitor chelerythrine (1 microM) reduced anterograde and retrograde axonal transport, while the CAM II kinase inhibitor KN-62 (10 microM) had no effect. Morphological observation showed that neurite growth was prevented by the presence of chelerythrine (1 microM). From these results, we conclude that PKC activity is required to maintain axonal transport and thereby neurite growth.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1999

Protein kinase C inhibition and activation phase advances the hamster circadian clock.

The mammalian circadian clock is located in the suprachiasmatic nuclei (SCN). Clock function can be detected by the measurement of the circadian change in cellular firing rate of SCN cells in vitro. We investigated the effects of protein kinase C (PKC) inhibition and activation on this rhythm of firing rate in hamster SCN neurons. PKC inhibition by chelerythrine chloride application phase advances the in vitro circadian rhythm during the late subjective night and early subjective morning, Zeitgeber time (ZT) 20-24 and ZT 0-4. No effect of PKC inhibition on clock phase was seen during ZT 6-18. Activation of PKC via phorbol 12-myristate 13-acetate (PMA) phase advanced the clock at all phases tested. Thus, at some circadian phases both inhibition and activation of PKC can advance circadian rhythms.

Alkaloids↗
PubMed2000

The effect of PKC activation on the survival of rat retinal ganglion cells in culture.

Natural cell death is a degenerative phenomenon occurring during the development of the nervous system. Approximately half the neurons initially generated during this period die. The role of trophic molecules produced by target and afferent neurons as well as by glial cells controlling this regressive event has been extensively demonstrated. The aim of this work was to study the role of activated protein kinase C (PKC), an enzyme involved in apoptosis regulation, on the survival of retinal ganglion cells kept "in vitro" for 48 h. For this purpose, we used the phorbol 12-myristate 13-acetate (PMA), a tumor promoter agent that activates PKC. Our results showed that PMA increases the survival of ganglion cells. The effect was dose-dependent and PMA concentrations of 10 or 100 ng/ml produced the maximal effect (a two-fold increase on ganglion cells survival compared with 48 h control). This effect was totally abolished by 1.25 microM chelerythrine chloride (an inhibitor of PKC) and 30 microM genistein (an inhibitor of tyrosine kinase enzymes). Otherwise, PMA was effective only when it was chronically present in the cultures. On the other hand, treatment with 20 microM 5-fluoro-2'-deoxyuridine, an inhibitor of cell proliferation, or 25 microM BAPTA-AM, an intracellular calcium chelator, did not block PMA effect. Our results suggest that the survival of retinal ganglion cells "in vitro" may be mediated by a mechanism that involves PKC activation.

Alkaloids↗
PubMed2000

Hypoxia differentially regulates stress proteins in cultured cardiomyocytes: role of the p38 stress-activated kinase signaling cascade, and relation to cytoprotection.

OBJECTIVE: Stress proteins (heat shock proteins, HSPs) are molecular chaperones that have been shown to enhance the survival of cells exposed to environmental stress. We sought to investigate the effects of hypoxia on the levels of HSP27 and heme oxygenase-1 (HO-1 or HSP32) in an established model of rat neonatal cardiac myocytes in culture. METHODS: Myocytes were subjected to hypoxia (<0.5% O(2) for 16 h). Studies of cell viability and nuclear morphology showed no evidence of cell death under these conditions. RESULTS: Messenger RNA analysis demonstrated constitutive expression of HSP27 and low levels of HO-1. Hypoxia strongly induced HO-1 mRNA without affecting HSP27 mRNA. In parallel to mRNA levels, hypoxia increased HO-1 protein level without affecting HSP27. To further assess the signaling pathways implicated in HO-1 induction, we used inhibition experiments. The tyrosine kinase inhibitor tyrphostin and the mitogen-activated protein kinase inhibitor PD98059 did not prevent HO-1 induction, while the protein kinase C inhibitor chelerythrine partially blocked this response. The p38 stress-activated kinase inhibitor SB203580 was the most potent in suppressing hypoxia-induced HO-1. In vitro kinase assays, cell labeling and immunoprecipitation showed activation of signaling pathways downstream of p38 stress-activated kinase as revealed by an increase in phosphorylation of MAPKAPK-2/3 kinases and HSP27. CONCLUSIONS: These data show a differential pattern of hypoxia-induced HSP expression and implicate the stress kinase in HO-1 induction. Thus, selective regulation of HSP levels may play a role in the cardioprotective mechanisms that participate in the adaptive response to hypoxia-induced stress.

Alkaloids↗
PubMed2001

Post-ischemic PKC inhibition impairs myocardial calcium handling and increases contractile protein calcium sensitivity.

OBJECTIVE: Protein kinase C (PKC) activation impairs contractility in the normal heart but is protective during myocardial ischemia. We hypothesized that PKC remains activated post-ischemia and modulates myocardial excitation-contraction coupling during early reperfusion. METHODS: Langendorff-perfused rabbit hearts where subjected to 25 min unmodified ischemia and 30 min reperfusion. Total PKC activity was measured, and the intracellular translocation pattern of PKC-alpha, -delta, -epsilon, and -eta assessed by immunohistochemistry and fractionated Western immunoblotting. The PKC-inhibitors chelerythrine and GF109203X were added during reperfusion and also given to non-ischemic hearts. Measurements included left ventricular function, intracellular calcium handling measured by Rhod-2 spectrofluorometry, myofibrillar calcium responsiveness in beating and tetanized hearts, and metabolic parameters. RESULTS: Total PKC activity was increased at end-ischemia and remained elevated after 30 min of reperfusion. The translocation pattern indicated PKC-epsilon as the main active isoform during reperfusion. Post-ischemic PKC inhibition affected mainly diastolic relaxation, with lesser effect on contractility. Both PKC inhibitors increased the Ca(2+) responsiveness of the myofilaments as indicated by a leftward shift of the calcium-to-force relationship and increased maximum calcium activated tetanic pressure. Diastolic Ca(2+) removal was delayed and the post-ischemic [Ca(2+)](i) overload further exacerbated. Depressed systolic function was associated with a lower amplitude of [Ca(2+)](i) transients. CONCLUSION: PKC is activated during ischemia and remains activated during early reperfusion. Inhibition of PKC activity post-ischemia impairs functional recovery, delays diastolic [Ca(2+)](i) removal, and increases Ca(2+) sensitivity of the contractile apparatus, resulting in impaired diastolic relaxation. Thus, post-ischemic PKC activity may serve to restore post-ischemic Ca(2+) homeostasis and attenuate contractile protein calcium sensitivity during the period of post-ischemic [Ca(2+)](i) overload.

Alkaloids↗
PubMed2002

Intestinal ischemia preconditions myocardium: role of protein kinase C and mitochondrial K(ATP) channel.

OBJECTIVE: The present study was designed to test the hypothesis that intestinal ischemia results in an early preconditioning against myocardial infarction and that the mechanism of the early preconditioning involves the activation of protein kinase C-mitochondrial K(ATP) channel signaling pathway in anesthetized rats. METHODS: Rats were either preconditioned with a 25-min occlusion of the superior mesenteric artery followed by 15 min of reperfusion or underwent a 40-min sham period. Subsequently, all rats were subjected to a sustained 30 min of coronary occlusion and 180 min of reperfusion. Infarct size was determined by triphenyltetrazolium chloride staining. RESULTS: In sham-operated rats receiving no pharmacological intervention, the percentage of myocardial infarct within the area at risk and left ventricle was 73+/-4% and 31+/-2%, respectively, and these were significantly reduced to 44+/-4% and 23+/-1% (P<0.01) after intestinal ischemia preconditioning. Intravenous injection of protein kinase C inhibitors chelerythrine (5 mg/kg) and staurosporine (50 microg/kg) or a specific mitochondrial K(ATP) channel inhibitor 5-hydroxydecanoate (5 mg/kg) 5 min before sustained myocardial ischemia abolished the preconditioning afforded by intestinal ischemia. However, hexamethonium, a ganglion blocker, did not attenuate the preconditioning. CONCLUSIONS: These data provide pharmacological evidence that protein kinase C and mitochondrial K(ATP) channel are involved in the mechanism of the early preconditioning induced by intestinal ischemia.

Alkaloids↗