Biomedical subjects

Shoei-Yn Lin-Shiau

Publications and source records attributed to Shoei-Yn Lin-Shiau.

At least 19 recordsLinked to original sources

PubMed2005

Comparative studies on the hypolipidemic and growth suppressive effects of oolong, black, pu-erh, and green tea leaves in rats.

The four major commercial teas, oolong, black, pu-erh, and green teas, have been manufactured in southeast Asia. In this study, we evaluated the growth suppressive and hypolipidemic effect of these four different tea leaves by oral feeding to male Sprague-Dawley rats for 30 weeks. The results showed that the suppression of body weights of tea leaves-fed groups were in the order: oolong tea > pu-erh tea > black tea > green tea. Pu-erh tea and oolong tea could lower the levels of triglyceride more significantly than that of green tea and black tea, but pu-erh tea and green tea were more efficient than oolong tea and black tea in lowering the level of total cholesterol. In lipoprotein, 4% pu-erh tea could increase the level of HDL-C and decrease the level of LDL-C, but other teas simply decrease the levels of both. The activity of antioxidant enzyme SOD is increased in all tea-fed groups as compared to the basal diet-fed group. Finally, relative weight ratios of liver to epididylmal adipose tissue were lower in feeding oolong tea and pu-erh tea groups. On the basis of these findings, it seemed that the fully fermented pu-erh and black tea leaves and partially fermented oolong tea leaves were more effective on their growth suppressive and hypolipidemic effects as compared to the nonfermented green tea leaves.

Animals↗

PubMed2004

Carnosol inhibits the invasion of B16/F10 mouse melanoma cells by suppressing metalloproteinase-9 through down-regulating nuclear factor-kappa B and c-Jun.

Carnosol, a constant constituent of Rosmarinus officinalis extracts, is a phenolic diterpene shown to have antioxidant and anticarcinogen properties. In our studies, carnosol inhibited the invasion of highly metastatic mouse melanoma B16/F10 cells in vitro. First, the antimetastatic potentials of carnosol were examined by soft agar colony formation assay. Second, carnosol dose-dependently inhibited B16/F10 cell migration and invasion by in vitro transwell assay. Third, the decreasing activity of metalloproteinase was observed by zymographic assay. The result revealed that the treatment of carnosol could diminish the activity of MMP-9 more than MMP-2. Next, we analyzed the amounts of MMP-9 and MMP-2 proteins in the cells. The data indicated MMP-9 protein was also suppressed by carnosol in the same manner. In accordance with the above data, the results of reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed a reduced level of MMP-9 mRNA. Furthermore, carnosol significantly inhibited the tyrosine phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, AKT, p38, JNK and inhibition of activation of transcription factors NFkappa-B and c-Jun. These results lead us to conclude that carnosol could restrict the invasive ability of B16/F10 mouse melanoma cells by reducing MMP-9 expression and activity through suppressing (ERK) 1/2, AKT, p38, and JNK signaling pathway and inhibition of NF-kappaB and AP-1 binding activity. Taken together, these results indicate that carnosol targets MMP-mediated cellular events in cancer cells and provides a new mechanism for its anticancer activity.

Abietanes↗

PubMed2004

Decrease in Ca2+-activated K+ conductance in differentiated C6-glioma cells.

Ca2+-activated K+ channels were studied in C6-glioma cells in an attempt to correlate changes in expression with cell proliferation and differentiation. In this study, we treated C6-glioma cells with thapsigargin for 48 h. Cell proliferation was markedly inhibited, and cell morphology changed from round to a spindle differentiated shape. Furthermore, intracellular calcium concentration was initially increased during acute treatment with thapsigargin. The internal [Ca2+]i pool was eventually depleted after a 48-h thapsigargin treatment. We have characterized Ca2+-activated K+ currents in less differentiated C6 cells. After differentiation of C6 cells induced by thapsigargin, Ca2+-activated K+ currents were selectively suppressed. These data lend further support to the notion that the expression of Ca2+-activated K+ channels is intimately associated with the proliferation of C6-glioma cells, and the suppression of Ca2+-activated K+ channels coincides with the inhibition of proliferation and subsequent induction of cell differentiation.

Animals↗

PubMed2004

Early acute necrosis and delayed apoptosis induced by methyl mercury in murine peritoneal neutrophils.

There is growing evidence that heavy metals in general, and mercurial compounds in particular, are immunotoxic. The purpose of this study was to explore the mechanism of MeHg in inducing cell death of mouse peritoneal neutrophils. In this paper we demonstrate that MeHg induces apoptosis and necrosis depending on MeHg concentration. In vitro exposure of mouse peritoneal neutrophils to MeHg resulted in a time- and concentration-dependent cell death. MeHg (15 microM) induced neutrophil necrosis in 13 min. The type of cell death was attributed to necrosis based on cells permeable to the fluorescent dye, propidium iodide and DNA appeared as a smear. With fura-2 microfluorimetric technique, we found that the entry of external Ca2+ into the cytosol played a crucial role in inducing cell necrosis by 15 microM MeHg. However, at lower concentrations, MeHg (10 microM)-induced apoptosis is confirmed by the observation of morphological features characterised by apoptotic bodies and fragmented DNA ladder. MeHg (10 microM) caused an immediate fall in pHi as revealed by the pH-sensitive fluorescent probe 2'7'-bis (carboxyethyl)-5(6)-carboxyfluorescein. We have found that MeHg induced cellular acidification prior to DNA fragmentation so as the other two apoptosis-inducing agents (ZnCl(2) and EGTA). Furthermore, acid-activated endonuclease was increased by MeHg in neutrophils, which we considered to play a possible role in chromatin digestion leading to apoptosis. Taken together, these findings indicate that MeHg induces necrosis at higher concentrations by a rapid increase of [Ca2+]i and apoptosis at lower concentrations by acid activation of endonuclease.

Animals↗

PubMed2003

The role of titanium in the altered endotoxin-induced nitric oxide synthase expression in alveolar macrophages from titanium-alloy-implanted rats.

We have found that the concentration of titanium (Ti) in the blood of patients with loosened Ti-alloy prostheses is elevated. An increase in the levels of elemental Ti in the blood and lung tissues of rats with an alloyed-Ti implant also has been found. The cellular reaction to elevated elemental Ti in the circulation remains unclear. We further performed experiments to examine the changes of inducible nitric oxide synthase (iNOS) expression in alveolar macrophages from alloyed-Ti-implanted rats. The elevation of nitrite and iNOS expression induced by lipopolysaccharide (LPS) was suppressed. The in vitro effect of a soluble form of Ti was further investigated. Ti (0.01-0.06 mM) inhibited the LPS-induced nitrite production and iNOS expression in alveolar macrophages from normal rats without any cytotoxic effects. LPS induced protein tyrosine phosphorylation, tyrosine-phosphorylation of lyn (a CD14-receptor-associated-tyrosine kinase), and degradation of IkappaB-alpha protein (inhibitor of NF-kappaB) in alveolar macrophages. These events were inhibited by co-incubation with Ti. These results indicate that elemental Ti may impair iNOS expression in alveolar macrophages through the alteration of protein tyrosine phosphorylation and NF-kappaB activation. The inhibitory action of Ti on cellular responses of alveolar macrophages may be anti-inflammatory and thus may depress local defense mechanisms related to microbial killing.

Alloys↗

PubMed2003

Motorcycle exhaust particulates enhance vasoconstriction in organ culture of rat aortas and involve reactive oxygen species.

The effects of motorcycle exhaust particulate on vasoconstriction were determined using rat thoracic aortas under organ culture conditions treated with organic extracts of motorcycle exhaust particulate from a two-stroke engine. The motorcycle exhaust particulate extract (MEPE) induced a concentration-dependent enhancement of vasoconstriction elicited by phenylephrine in the organ cultures of both intact and endothelium-denuded aortas for 18 h. Nifedipine (an L-type Ca2+ channel blocker), manganese acetate (an inorganic Ca2+ channel blocker), and staurosporine (a nonselective protein kinase C inhibitor), but not the selective protein kinase C inhibitor chelerythrine, inhibited the enhancement of vasoconstriction by MEPE. Staurosporine has also been reported as a myosin light chain kinase (MLCK) inhibitor, so we tested whether the MLCK pathway was involved in the effect of MEPE. The results showed that ML-9 (a selective MLCK inhibitor) could inhibit the enhancement of vasoconstriction by MEPE. The phosphorylation of a 20-kDa myosin light chain in a primary culture of rat vascular smooth muscle cells was also enhanced by MEPE. Moreover, we also examined the role of reactive oxygen species (ROS) in the stimulatory effect of MEPE on vasoconstriction. The antioxidant N-acetylcysteine significantly inhibited the enhancement of vasoconstriction by MEPE. A time-dependent increase in ROS production by MEPE was also detected in primary cultures of vascular smooth muscle cells. These results indicate that MEPE induces a marked enhancement of vasoconstriction in aortas under organ culture conditions and imply that a ROS-Ca2+-MLCK pathway may be involved in this MEPE-induced response.

Air Pollutants↗

PubMed2003

Suramin inhibits beta-bungarotoxin-induced activation of N-methyl-D-aspartate receptors and cytotoxicity in primary neurons.

We demonstrated that beta-bungarotoxin (beta-BuTX), a snake presynaptic neurotoxin, exhibited a potent cytotoxic effect on cultured cerebellar granule neurons. The mechanism of action of beta-BuTX and the cytoprotective agents against beta-BuTX were studied. The neuronal death of cerebellar granule neurons induced by beta-BuTX was manifested with apoptosis and necrosis processes as revealed by neurite fragmentation, morphological alterations, and staining apoptotic bodies with the fluorescent dye Hoechst 33258. By means of microspectrofluorimetry and fura-2, we measured intracellular Ca2+ concentration, [Ca2+]i and found that [Ca2+]i was increased markedly prior to the morphological changes and cytotoxicity. The downstream pathway of the increased [Ca2+]i was investigated: there was increased production of free radicals, decreased mitochondrial membrane potential, and depleted cellular ATP content. MK801 and suramin effectively suppressed these detrimental effects of beta-BuTX. Furthermore, the [3H]MK801 binding was reduced by unlabeled MK801, beta-BuTX, and suramin. Thus, activation of N-methyl-D-aspartate (NMDA) receptors appeared to play a crucial role in the cytotoxic effects following betaBuTX exposure. In conclusion, the novel finding of this study was that a polypeptide beta-BuTX exerted a potent cytotoxic effect through sequential events, including activating NMDA receptors followed by increasing [Ca2+]i, ROS production, and impaired mitochondrial energy metabolism. Suramin, clinically used as a trypanocidal agent, was an effective antagonist against beta-BuTX. Data suggest that suramin might have value to detect the possible pathway of certain neuropathological disorders.

Animals↗

PubMed2003

beta-Lapachone reduces endotoxin-induced macrophage activation and lung edema and mortality.

beta-Lapachone, a 1,2-naphthoquinone, is a novel chemotherapeutic agent. It has been shown to be capable of suppressing inducible nitric oxide synthase expression and function in rat alveolar macrophages. The authors further performed experiments to examine the molecular mechanism of beta-lapachone on LPS-induced responses in rat alveolar macrophages and to evaluate its in vivo antiinflammatory effect. A significant increase in nitrite production and inducible nitric oxide synthase expression was elicited in macrophages treated with LPS that was inhibited by coincubation with beta-lapachone. beta-Lapachone could also inhibit the production of tumor necrosis factor-alpha induced by LPS. LPS induces protein tyrosine phosphorylation and nuclear factor-kappaB binding activity by gel mobility shift assay in macrophages. These events were significantly inhibited by beta-lapachone. Furthermore, beta-lapachone in vivo protected against the induction of lung edema, lung-inducible nitric oxide synthase protein expression and nuclear factor-kappaB activation, lethality, and increased plasma nitrite and serum tumor necrosis factor-alpha levels induced by LPS. These results indicate that beta-lapachone suppresses inducible nitric oxide synthase induction and tumor necrosis factor-alpha production mediated by the inhibition of protein tyrosine phosphorylation and nuclear factor-kappaB activation caused by LPS. This results in a beneficial effect in an animal model of sepsis.

Animals↗

PubMed2003

Calcium-activated NO production plays a role in neuronal death induced by beta-bungarotoxin in primary cultures of cerebellar granular neurons.

The aim of this study was to elucidate the mechanism underlying the neurotoxic effect of beta-bungarotoxin (beta-BuTX) on cultured cerebellar granular neurons (CGN). Beta-BuTX had a potent time- and concentration-dependent neurotoxic effect on mature CGN. Beta-BuTX appeared to destroy initially the neurites and then caused neuronal death by both apoptotic and necrotic processes. Inspection using Nomarski optics showed that these neurons displayed morphological features of necrotic cells, including cell swelling, loss of membrane integrity and eventual dissolution of the cell. Staining with the fluorescent dye Hoechst 33258 showed that beta-BuTX-treated neuron bodies stained more densely with smaller apoptotic bodies. Using microspectrofluorimetry and fura-2 to measure cytosolic [Ca(2+)] ([Ca(2+)](i)), beta-BuTX markedly increased [Ca(2+)](i). BAPTA-AM, EGTA, MK 801 and diltiazem not only attenuated the beta-BuTX-mediated rise in [Ca(2+)](i) but also attenuated beta-BuTX-mediated neurotoxicity. In addition, these Ca(2+) inhibitors prevented the beta-BuTX-induced generation of reactive nitrogen species. The NO synthase inhibitor N(G)-methyl- l-arginine) also exhibited neuroprotection. This is the first report showing that beta-BuTX-induced CGN death is mediated, at least in part, by excessive generation of NO triggered by [Ca(2+)](i) overloading. Activation of NMDA receptors and L-type calcium channels is apparently involved in the increase in [Ca(2+)](i) induced by this neurotoxin. This potent neurotoxin will be a useful tool for studying neurotoxic processes and using this model system will allow us to find neuroprotective agents.

Animals↗

PubMed2003

Resveratrol-induced G2 arrest through the inhibition of CDK7 and p34CDC2 kinases in colon carcinoma HT29 cells.

Resveratrol (3,5,4'-trihydroxystilbene), a phytoalexin found in grapes and other food products, has been shown to have cancer chemopreventive activity. However, the mechanism of the anti-carcinogenic activity is not well understood. Here, we offer a possible explanation of its anti-tumor effect. Based on flow cytometric analysis, resveratrol inhibited the proliferation of HT29 colon cancer cells and resulted in their accumulation in the G(2) phase of the cell cycle. Western blot analysis and kinase assays demonstrated that the perturbation of G(2) phase progression by resveratrol was accompanied by the inactivation of p34(CDC2) protein kinase, and an increase in the tyrosine phosphorylated (inactive) form of p34(CDC2). Kinase assays revealed that the reduction of p34(CDC2) activity by resveratrol was mediated through the inhibition of CDK7 kinase activity, while CDC25A phosphatase activity was not affected. In addition, resveratrol-treated cells were shown to have a low level of CDK7 kinase-Thr(161)-phosphorylated p34(CDC2). These results demonstrated that resveratrol induced cell cycle arrest at the G(2) phase through the inhibition of CDK7 kinase activity, suggesting that its anti-tumor activity might occur through the disruption of cell division at the G(2)/M phase.

Antineoplastic Agents↗

PubMed2003

Neuronal death signaling by beta-bungarotoxin through the activation of the N-methyl-D-aspartate (NMDA) receptor and L-type calcium channel.

The aim of this study was to elucidate the mechanism of the neurotoxic effect of beta-bungarotoxin (beta-BuTX, a snake presynaptic neurotoxin isolated from the venom of Bungarus multicinctus) on cultured cerebellar granule neurons. beta-BuTX exerted a potent, time-dependent, neurotoxic effect on mature granule neurons. Mature neurons, with an abundance of neurite outgrowths, were obtained after 7-8 days in culture. By means of microspectrofluorimetry and fura-2, we measured the intracellular Ca(2+) concentration ([Ca(2+)](i)) and found it to be increased markedly. BAPTA-AM [1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tertrakis(acetoxymethyl ester)], EGTA, MK801 (dizocilpine maleate), and diltiazem prevented not only the elevation of [Ca(2+)](i), but also the beta-BuTX-induced neurotoxic effect. The signaling pathway involved in the elevation of [Ca(2+)](i) in beta-BuTX-induced neurotoxicity was studied. The results obtained indicated that beta-BuTX initially increased the production of reactive oxygen species and subsequently reduced mitochondrial membrane potential and depleted ATP. All of these events in the signaling pathway were blocked by MK801, diltiazem, EGTA, and BAPTA-AM. These findings suggest that the neurotoxic effect of beta-BuTX is mediated, at least in part, by a cascade of events that include the direct or indirect activation of N-methyl-D-aspartate (NMDA) receptors and L-type calcium channels that, in turn, lead to Ca(2+) influx, oxidative stress, mitochondrial dysfunction, and ATP depletion. Therefore, we suggest that this polypeptide neurotoxin, as a result of its high potency and irreversible properties, is a useful tool to elucidate the mechanisms of neurodegenerative diseases.

Adenosine Triphosphate↗

PubMed2003

The clinical significance of venom detection in patients of cobra snakebite.

Cobra snakebites are one of the most frequent occurrences among the poisonous snakebites. The correlation between the serum concentrations of cobra venom in the victims with the severity of systemic and local symptoms after envenoming is still awaited for investigation. In this paper, we have developed an enzyme-linked immunosorbent assay (ELISA) to detect Taiwan cobra venom existed in the serum of the cobra snakebite victims. This established technique allowed us to detect as low as 1 ng/ml. Totally, 31 serum samples of 27 recognized and suspected cobra snakebite patients were analyzed by the developed ELISA technique. The highest venom level detected was 1270 ng/ml in a case 2 h after being envenomed. We found that the serum concentrations of the venom in the bitten patients were well correlated with the severity of local tissue destruction. Venom levels might remain high 48 h after snakebite. Two or more vials of specific antivenom administered soon after snakebite were effective in neutralizing the circulating venom as revealed by ELISA. Higher doses of antivenom may be indicated in cases of cobra venom-induced local tissue destruction. In conclusion, these results indicate that venom detection in snakebite victims is valuable in diagnosis and severity assessment of Taiwan cobra snakebite.

Adolescent↗

PubMed2003

Activation of NMDA receptor partly involved in beta-bungarotoxin-induced neurotoxicity in cultured primary neurons.

In this study, we demonstrated that a snake presynaptic toxin, beta-bungarotoxin (beta-BuTX), was capable of binding to NMDA receptors of the cultured primary neurons (cerebellar granule neurons, CGNs). We labeled beta-BuTX with fluorescent FITC (FITC-beta-BuTX) and showed that the binding of FITC-beta-BuTX was inhibited by unlabeled beta-BuTX and MK801 (an NMDA receptor antagonist). Meanwhile, the binding of [3H]-MK801 was also reduced by unlabeled MK801 and beta-BuTX. In addition, beta-BuTX produced a very potent neurotoxic effect on mature CGNs with the EC(50) of 3ng/ml (equivalent to 144pM), but was less effective in immature CGNs. We explored the signaling pathway of neuronal death and found that it was apparently due to the excessive production of reactive oxygen species (ROS) induced by beta-BuTX. MK801 and antioxidants (Vitamin C, N-acetylcysteine (NAC), melatonin, epigallocatechin gallate (EGCG), superoxide dismutase (SOD) and catalase) attenuated not only ROS production but also beta-BuTX-neurotoxicity. The downstream signaling of ROS was identified as the activation of caspase-3. Caspase inhibitor (z-DEVD-fmk) and antioxidants depressed both caspase-3 activation and neurotoxicity. Based on these findings and our previous reports, we conclude that the binding and activation of NMDA receptors by beta-BuTX was crucial step to produce the potent neurotoxic effect. The binding of NMDA receptors resulted in excessive Ca(2+) influx, followed by ROS production and activation of caspase-3. This snake toxin is considered not only to be a useful tool for exploring the death-signaling pathway of neurotoxicity, but also provides a model for searching neuroprotective agents.

Animals↗

PubMed2003

The effect of L-arginine on slow motility of mammalian outer hair cell.

The effect of L-arginine on the slow motility of mammalian cochlear outer hair cells was studied in this experiment. L-Arginine (3 mM) but not D-arginine (3 mM) or other amino acids (L-aspartate or L-glutamate) induced length increases of guinea pig outer hair cell. Similarly, the membrane-permeant cGMP analogues, 8-(4-chlorophenylthio)guanosine 3':5'-cyclic monophosphate (1 mM) or 8-bromo-guanosine 3':5'-cyclic monophosphate (1 mM) induced length increases of guinea pig outer hair cells. These length increases induced by L-arginine can be attenuated by a 30 min preincubation of the cells with the nitric oxide synthase inhibitors N(G)-nitro-L-arginine methyl ester hydrochloride (3 mM) or 7-nitroindazole (1 mM). Comparing the effects of L-arginine and ionomycin on cell length and intracellular calcium change in outer hair cells, both L-arginine and ionomycin were able to induce the elongation of outer hair cells but L-arginine did not change the fluorescence intensity of Fluo-3. Preincubation of the cells with EGTA (3 mM) for 40 min to reduce the extracellular calcium concentration did not influence the effect of L-arginine. This experiment demonstrated that nitric oxide/cGMP pathway involvement in regulating the slow motility of mammalian outer hair cells cannot be ruled out. The effect of L-arginine is independent of extracellular calcium concentration.

Animals↗

PubMed2002

Penta-O-galloyl-beta-D-glucose inhibits the invasion of mouse melanoma by suppressing metalloproteinase-9 through down-regulation of activator protein-1.

Penta-O-galloyl-beta-D-glucose (5GG) inhibited the invasion of highly metastatic mouse melanoma B16F10 cells in vitro, as demonstrated by transwell assay. Its ability to diminish the activity of matrix metalloproteinase (MMP) was demonstrated by zymographic assay. Our data showed 5GG could diminish the activity of MMP-9 more than that of MMP-2. The effect on MMP-9 was elicited in a dose- and time-dependent manner, with IC50 of 15 microM. Next, we analyzed the amounts of MMP-9 and MMP-2 protein in conditioned media and in the cells. The data indicated MMP-9 proteins were also suppressed by 5GG in the same manner. In accordance with these data above, the results of reverse transcriptase polymerase chain reaction (RT-PCR) and Northern blot analysis showed a reduced level of MMP-9 mRNA. Furthermore, we studied transcription factor binding to MMP-2 and MMP-9 promoter regions by electrophoretic mobility shift assay (EMSA) in the nucleus. The results suggested that the transcription factor binding activities of Activator protein-1 (AP-1) and Sp-1 sites was mainly down-regulated by 5GG in the concentration range of 5-15 microM, but not that of nuclear factor kappaB (NF-kappaB), polioma enhancer activator 3 (PEA-3), and Activator protein-2 (AP-2) sites. The Western blot analysis of AP-1 nuclear protein showed a reduced level of c-Jun but not of c-Fos. In addition, the expression of Sp-1 and c-Jun protein was also suppressed. To elucidate whether the transcriptional activity of AP-1 or Sp-1 sites is more important, we transfected MMP-9/luciferase reporter vector, under MMP-9 promoter control, into the cells. We found that a decreased transcriptional activity of AP-1 sites is sufficient to reduce MMP-9 promoter activity. These results lead us to conclude that 5GG restricts the invasive ability of B16F10 mouse melanoma cells by reducing MMP-9 activity, by suppressing the transcriptional activity of AP-1 sites and the expression of c-Jun protein. The result may provide a potential mechanism for 5GG in cancer chemopreventive action.

Animals↗

PubMed2002

Long-term lithium treatment prevents neurotoxic effects of beta-bungarotoxin in primary cultured neurons.

Lithium is the most commonly used drug for the treatment of manic-depressive illness. The precise mechanisms underlying its clinical efficacy remain unknown. In this study, we found that long-term exposure to lithium chloride protected cultured cerebellar granule neurons (CGNs) against beta-bungarotoxin (beta-BuTX)-induced neurotoxicity. This neuroprotection was exhibited at the therapeutically relevant concentration of 1.2 mM lithium. Pretreatments for 3-5 days (long-term) were required for protection to occur; but a 3 hr treatment (short-term) was ineffective. In contrast, a longer treatment for 6-7 days or a higher concentration of 3 mM lithium led not only to loss of the neuroprotective effect but also to a neurotoxic effect. These findings suggest that lithium protection is limited to its narrow window of concentration and apparently relevant to its narrow therapeutic index in clinical application. Measurement of intracellular calcium [Ca(2+)](i) revealed that neurotoxic concentrations of beta-BuTX markedly increased [Ca(2+)](i), which could be attenuated by long-term, but not short-term, lithium treatment. Thus, the protection induced by lithium in CGNs was attributed to its inhibition of calcium overload. In addition, the Ca(2+) signaling pathway, including reactive oxygen species production and mitochondrial membrane potential reduction, along with the neurotoxic effect of beta-BuTX was blocked by long-term, but not short-term, lithium treatment. All of these results indicate that a crucial step for lithium protection is modulation of [Ca(2+)](i) homeostasis and that lithium neurotoxicity possibly, at least in part, is due to calcium overload. In conclusion, our results suggest that lithium, in addition to its use in treatment of bipolar depressive illness, may have an expanded use in intervention for neurotoxicity.

Analysis of Variance↗

PubMed2002

Nitric oxide is not involved in the endotoxemia-induced alterations in Ca2+ and ryanodine responses in mouse diaphragms.

Lipopolysaccharide (LPS, endotoxin)-induced diaphragmatic contractile dysfunction and sarcolemmal injury in animals has been identified. However, the precise nature of sepsis-related alterations in diaphragm myofiber function and the activity of Ca(2+) release from sarcoplasmic reticulum of skeletal muscle remain unclear. The present study investigated the in vivo effects of LPS on the Ca(2+)-dependent mechanical activity and ryanodine response in mouse diaphragm and Ca(2+) release from isolated sarcoplasmic reticulum membrane vesicles, and aimed to examine the role of nitric oxide (NO) in these responses. When diaphragms were bathed in a solution that was Cl(-)-free, Na(+)-free, but contained high K(+), a Ca(2+)-induced contracture was elicited. Increases in external Ca(2+) concentration produced increases in peak tension of Ca(2+)-induced contracture in control diaphragm, while a decrease was seen in endotoxemic diaphragm. Ryanodine induced a marked contracture in control diaphragms, which was diminished after endotoxemia. This finding is correlated with the decrease of ryanodine-induced Ca(2+) release and the suppression of [(3)H]ryanodine binding on the isolated SR of the skeletal muscle from LPS-treated rats. In mice treated with LPS significantly increased levels of plasma nitrite and serum TNF-alpha were observed, changes inhibited by aminoguanidine [an inhibitor of inducible NO synthase (iNOS)] and pentoxifylline (an inhibitor of tumor necrosis factor-alpha formation), respectively. Moreover, LPS treatment resulted in a significant expression of mRNA for iNOS in mouse diaphragms. The inhibitory effects on Ca(2+)- and ryanodine responses by LPS could be prevented by treatment with polymyxin B (LPS neutralizer) and pentoxifylline, but not by treatment with dexamethasone, N(G)-nitro- L-arginine or aminoguanidine (NOS inhibitors). These results imply that the NO-related pathway may not be involved in the dysfunction of the Ca(2+) release mechanism in the sarcoplasmic reticulum of mouse diaphragm during endotoxemia.

Animals↗

PubMed2002

Methylmercury inhibits nitric oxide production mediated by Ca(2+) overload and protein kinase A activation.

The importance of cytosolic free calcium level intracellular Ca(2+), [Ca(2+)]i, in neutrophil activation prompted us to investigate changes in [Ca(2+)]i of neutrophils caused by methylmercury (MeHg), which has been shown to have immunomodulatory properties. We have shown in this paper that MeHg increased [Ca(2+)]i in the mouse peritoneal neutrophil. The L-type calcium channel blocker verapamil can decrease the elevated [Ca(2+)]i caused by 10 microM MeHg, suggesting that Ca(2+)-influx through L-type Ca(2+) channel mediates the effect of MeHg. Moreover, MeHg potently decreased nitric oxide (NO) production but also the protein and mRNA level of NO synthase induced by lipopolysaccharide. Both verapamil (1 microM) and H-89 (10 microM) can antagonize the inhibitory effect of MeHg (10 microM) on NO production. These findings lead us to conclude that MeHg inhibits NO production mediated at least in part by Ca(2+)-activated adenylate cyclase-cAMP-protein kinase A pathway.

Animals↗