AIM:To investigate the effect of diazoxide administration on liver ischemia/reperfusion injury.METHODS:Wistar male rats underwent partial liver ischemia performed by clamping the pedicle from the medium and left anter...AIM:To investigate the effect of diazoxide administration on liver ischemia/reperfusion injury.METHODS:Wistar male rats underwent partial liver ischemia performed by clamping the pedicle from the medium and left anterior lateral segments for 1 h under mechanical ventilation.They were divided into 3 groups:Control Group,rats submitted to liver manipulation,Saline Group,rats received saline,and Diazoxide Group,rats received intravenous injection diazoxide(3.5 mg/kg) 15 min before liver reperfusion.4 h and 24 h after reperfusion,blood was collected for determination of aspartate transaminase(AST),alanine transaminase(ALT),tumor necrosis factor(TNF-α),interleukin-6(IL-6),interleukin-10(IL-10),nitrite/nitrate,creatinine and tumor growth factor-β1(TGF-β1).Liver tissues were assembled for mitochondrial oxidation and phosphorylation,malondialdehyde(MDA) content,and histologic analysis.Pulmonary vascular permeability and myeloperoxidase(MPO) were also determined.RESULTS:Four hours after reperfusion the diazoxide group presented with significant reduction of AST(2009 ± 257 U/L vs 3523 ± 424 U/L,P = 0.005); ALT(1794 ± 295 U/L vs 3316 ± 413 U/L,P = 0.005); TNF-α(17 ± 9 pg/mL vs 152 ± 43 pg/mL,P = 0.013; IL-6(62 ± 18 pg/mL vs 281 ± 92 pg/mL); IL-10(40 ± 9 pg/mL vs 78 ± 10 pg/mL P = 0.03),and nitrite/nitrate(3.8 ± 0.9 μmol/L vs 10.2 ± 2.4 μmol/L,P = 0.025) when compared to the saline group.A significant reduction in liver mitochondrial dysfunction was observed in the diazoxide group compared to the saline group(P < 0.05).No differences in liver MDA content,serum creatinine,pulmonary vascular permeability and MPO activity were observed between groups.Twenty four hours after reperfusion the diazoxide group showed a reduction of AST(495 ± 78 U/L vs 978 ± 192 U/L,P = 0.032); ALT(335 ± 59 U/L vs 742 ± 182 U/L,P = 0.048),and TGF-β1(11 ± 1 ng/mL vs 17 ± 0.5 ng/mL,P = 0.004) serum levels when compared to the saline group.The control group did not present alterations when compared to the diazoxide and saline groups.CONCLUSION:Diazoxide maintains liver mitochondrial function,increases liver tolerance to ischemia/reperfusion injury,and reduces the systemic inflammatory response.These effects require further evaluation for using in a clinical setting.展开更多
Hypoxic pulmonary hypertension(HPH) is a syndrome characterized by the increase of pulmonary vascular tone and the structural remodeling of peripheral pulmonary arteries.The aim of specific therapies for hypoxic pulmo...Hypoxic pulmonary hypertension(HPH) is a syndrome characterized by the increase of pulmonary vascular tone and the structural remodeling of peripheral pulmonary arteries.The aim of specific therapies for hypoxic pulmonary hypertension is to reduce pulmonary vascular resistance,reverse pulmonary vascular remodeling,and thereby improving right ventricular function.Iptakalim,a lipophilic para-amino compound with a low molecular weight,has been demonstrated to be a new selective ATP-sensitive potassium(K ATP) channel opener via pharmacological,electrophysiological,biochemical studies,and receptor binding tests.In hypoxia-induced animal models,iptakalim decreases the elevated mean pressure in pulmonary arteries,and attenuates remodeling in the right ventricle,pulmonary arteries and airways.Furthermore,iptakalim has selective antihypertensive effects,selective vasorelaxation effects on smaller arteries,and protective effects on endothelial cells,but no effects on the central nervous,respiratory,digestive or endocrine systems at therapeutic dose.Our previous studies demonstrated that iptakalim inhibited the effects of endothelin-1,reduced the intracellular calcium concentration and inhibited the proliferation of pulmonary artery smooth muscle cells.Since iptakalim has been shown safe and effective in both experimental animal models and phase I clinical trials,it can be a potential candidate of HPH in the future.展开更多
Increasing evidence, including from our laboratory, has revealed that opening of ATP sensitive potassium channels(K-ATP channels) plays the neuronal protective roles both in vivo and in vitro. Thus K-ATP channel opene...Increasing evidence, including from our laboratory, has revealed that opening of ATP sensitive potassium channels(K-ATP channels) plays the neuronal protective roles both in vivo and in vitro. Thus K-ATP channel openers(KCOs) have been proposed as potential neuroprotectants. Our previous studies demonstrated that K-ATP channels could regulate glutamate uptake activity in PC12 cells as well as in synaptosomes of rats. Since glutamate transporters(GluTs) of astrocytes play crucial roles in glutamate uptake and KATP channels are also expressed in astrocytes, the present study showed whether and how KATP channels regulated the function of GluTs in primary cultured astrocytes. The results showed that nonselective KCO pinacidil, selective mitochondrial KCO diazoxide, novel, and blood-brain barrier permeable KCO iptakalim could enhance glutamate uptake, except for the sarcolemmal KCO P1075. Moreover pinacidil, diazoxide, and iptakalim reversed the inhibition of glutamate uptake induced by 1-methyl-4-phenylpyridinium(MPP+). These potentiated effects were completely abolished by mitochondrial K-ATP blocker 5-hydroxydecanoate. Furthermore, either diazoxide or iptakalim could inhibit MPP+-induced elevation of reactive oxygen species (ROS) and phosphorylation of protein kinases C(PKC). These findings are the first to demonstrate that activation of K-ATP channel, especially mitochondrial K-ATP channel, improves the function of GluTs in astrocytes due to reducing ROS production and downregulating PKC phosphorylation. Therefore, the present study not only reveals a novel pharmacological profile of KCOs as regulators of GluTs, but also provides a new strategy for neuroprotection.展开更多
AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclea...AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)pathway on such an effect.METHODS Male Wistar rats were randomly divided into a control group,a physiological saline(PS)group,a sodium hydrosulfide(Na HS)group,a glibenclamide(Gl)group,Gl plus Na HS group,a pyrrolidine dithiocarbamate(PDTC)group,and a PDTC plus Na HS group.Gastric mucosal injury was induced by RWIS for 3 h in rats,and gastric mucosal damage was analyzed after that.The PS,Na HS(100μmol/kg body weight),Gl(100μmol/kg body weight),Gl(100μmol/kg or 150μmol/kg body weight)plus Na HS(100μmol/kg body weight),PDTC(100μmol/kg body weight),and PDTC(100μmol/kg body weight)plus Na HS(100μmol/kg bodyweight)were respectively injected intravenously before RWIS.RESULTS RWIS induced serious gastric lesions in the rats in the PS pretreatment group.The pretreatment of Na HS(a H2S donor)significantly reduced the damage induced by RWIS.The gastric protective effect of the Na HS during RWIS was attenuated by PDTC,an NF-κB inhibitor,and also by glibenclamide,an ATP-sensitive potassium channel blocker,in a dose-dependent manner.CONCLUSION These results suggest that exogenous H2S plays a protective role against RWIS injury in rats,possibly through modulation of KATP channel opening and the NF-κB dependent pathway.展开更多
Objective:To investigate the role of iptakalim,an ATP-sensitive potassium channel opener,in transient cerebral ischemia/reperfusion (I/R) injury and its involved mechanisms.Methods:Intraluminal occlusion of middle cer...Objective:To investigate the role of iptakalim,an ATP-sensitive potassium channel opener,in transient cerebral ischemia/reperfusion (I/R) injury and its involved mechanisms.Methods:Intraluminal occlusion of middle cerebral artery (MCAO) in a rat model was used to investigate the effect of iptakalim at different time points.Infarct volume was measured by staining with 2,3,5-triphenyltetrazolium chloride,and immunohistochemistry was used to evaluate the expressions of Bcl-2 and Bax.In vitro,neurovascular unit (NVU) cells,including rat primary cortical neurons,astrocytes,and cerebral microvascular endothelial cells,were cultured and underwent oxygen-glucose deprivation (OGD).The protective effect of iptakalim on NVU cells was investigated by cell viability and injury assessments,which were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and release of lactate dehydrogenase.Caspase-3,Bcl-2 and Bax mRNA expressions were evaluated by real-time polymerase chain reaction (PCR).Results:Administration of iptakalim 0 or 1 h after reperfusion significantly reduced infarct volumes,improved neurological scores,and attenuated brain edema after cerebral I/R injury.Iptakalim treatment (0 h after reperfusion) also reduced caspase-3 expression and increased the ratio of Bcl-2 to Bax by immunohistochemistry.Iptakalim inhibited OGD-induced cell death in cultured neurons and astrocytes,and lactate dehydrogenase release from cerebral microvascular endothelial cells.Iptakalim reduced mRNA expression of caspase-3 and increased the ratio of Bcl-2 to Bax in NVU cells.Conclusions:Iptakalim confers neuroprotection against cerebral I/R injury by protecting NVU cells via inhibiting of apoptosis.展开更多
Cumulative evidence suggests that renal vascular endothelial injury play an important role in initiating and extending tubular epithelial injury and contribute to the development of ischemic acute renal failure.Our pr...Cumulative evidence suggests that renal vascular endothelial injury play an important role in initiating and extending tubular epithelial injury and contribute to the development of ischemic acute renal failure.Our previous studies have demonstrated that iptakalim's endothelium protection is related to activation of SUR2B/Kir6.1 subtype of ATP sensitive potassium channel(K ATP) in the endothelium.It has been reported that SUR2B/Kir6.1 channels are widely distributed in the tubular epithelium,glomerular mesangium,and the endothelium and the smooth muscle of blood vessels.Herein,we hypothesized that activating renal K ATP channels with iptakalim might have directly neroprotective effects.In this study,glomerular endothelial,mesangial and tubular epithelial cells which are the main cell types to form nephron were exposed to oleic acid(OA) at various concentrations for 24 h.0.25 μl/ml OA could cause cellular damage of glomerular endothelium and mesangium,while 1.25μl/ml OA could lead to the injury of three types of renal cells.It was observed that pretreatment with iptakalim at concentrations of 0.1,1,10 or 100 μmol/L prevented cellular damage of glomerular endothelium and tubular epithelium,whereas iptakalim from 1 to 100 μmol/L prevented the injury of mesangial cells.Our data showed iptakalim significantly increased survived cell rates in a concentration-dependent manner,significantly antagonized by glibenclamide,a K ATP blocker.Iptakalim played a protective role in the main cell types of kidney,which was consistent with natakalim,a highly selective SUR2B/Kir6.1 channel opener.Iptakalim exerted protective effects through activating SUR2B/Kir6.1 channels,suggesting a new strategy for renal injury by its endothelial and renal cell protection.展开更多
ATP-sensitive potassium channel(KATP) consists of a 4.4 complex of an inwardly rectifying Kir6.x pore plus a sulfonylurea receptor,which is an ATP-binding cassette transporter.KATP has been indentified in a variety of...ATP-sensitive potassium channel(KATP) consists of a 4.4 complex of an inwardly rectifying Kir6.x pore plus a sulfonylurea receptor,which is an ATP-binding cassette transporter.KATP has been indentified in a variety of tissues and recognized as an important drug target.It connects cell metabolism with cell electric activity.KATP has been proposed to play protective roles during heart failure,arrhythmia,myocardial infarction,stress,myocardial ischemia and hypertension.In this review,a summary of KATP is presented with molecular structure,localization,regulation,cardiovascular protective effect and its mechanisms.展开更多
基金Supported by Fundao de Amparoà Pesquisa do Estado de So Paulo,No.2010/19078-1
文摘AIM:To investigate the effect of diazoxide administration on liver ischemia/reperfusion injury.METHODS:Wistar male rats underwent partial liver ischemia performed by clamping the pedicle from the medium and left anterior lateral segments for 1 h under mechanical ventilation.They were divided into 3 groups:Control Group,rats submitted to liver manipulation,Saline Group,rats received saline,and Diazoxide Group,rats received intravenous injection diazoxide(3.5 mg/kg) 15 min before liver reperfusion.4 h and 24 h after reperfusion,blood was collected for determination of aspartate transaminase(AST),alanine transaminase(ALT),tumor necrosis factor(TNF-α),interleukin-6(IL-6),interleukin-10(IL-10),nitrite/nitrate,creatinine and tumor growth factor-β1(TGF-β1).Liver tissues were assembled for mitochondrial oxidation and phosphorylation,malondialdehyde(MDA) content,and histologic analysis.Pulmonary vascular permeability and myeloperoxidase(MPO) were also determined.RESULTS:Four hours after reperfusion the diazoxide group presented with significant reduction of AST(2009 ± 257 U/L vs 3523 ± 424 U/L,P = 0.005); ALT(1794 ± 295 U/L vs 3316 ± 413 U/L,P = 0.005); TNF-α(17 ± 9 pg/mL vs 152 ± 43 pg/mL,P = 0.013; IL-6(62 ± 18 pg/mL vs 281 ± 92 pg/mL); IL-10(40 ± 9 pg/mL vs 78 ± 10 pg/mL P = 0.03),and nitrite/nitrate(3.8 ± 0.9 μmol/L vs 10.2 ± 2.4 μmol/L,P = 0.025) when compared to the saline group.A significant reduction in liver mitochondrial dysfunction was observed in the diazoxide group compared to the saline group(P < 0.05).No differences in liver MDA content,serum creatinine,pulmonary vascular permeability and MPO activity were observed between groups.Twenty four hours after reperfusion the diazoxide group showed a reduction of AST(495 ± 78 U/L vs 978 ± 192 U/L,P = 0.032); ALT(335 ± 59 U/L vs 742 ± 182 U/L,P = 0.048),and TGF-β1(11 ± 1 ng/mL vs 17 ± 0.5 ng/mL,P = 0.004) serum levels when compared to the saline group.The control group did not present alterations when compared to the diazoxide and saline groups.CONCLUSION:Diazoxide maintains liver mitochondrial function,increases liver tolerance to ischemia/reperfusion injury,and reduces the systemic inflammatory response.These effects require further evaluation for using in a clinical setting.
基金supported by National Major Scientific and Technological Special Project for"Significant New Drugs Development"(2011ZX09302-003-02)
文摘Hypoxic pulmonary hypertension(HPH) is a syndrome characterized by the increase of pulmonary vascular tone and the structural remodeling of peripheral pulmonary arteries.The aim of specific therapies for hypoxic pulmonary hypertension is to reduce pulmonary vascular resistance,reverse pulmonary vascular remodeling,and thereby improving right ventricular function.Iptakalim,a lipophilic para-amino compound with a low molecular weight,has been demonstrated to be a new selective ATP-sensitive potassium(K ATP) channel opener via pharmacological,electrophysiological,biochemical studies,and receptor binding tests.In hypoxia-induced animal models,iptakalim decreases the elevated mean pressure in pulmonary arteries,and attenuates remodeling in the right ventricle,pulmonary arteries and airways.Furthermore,iptakalim has selective antihypertensive effects,selective vasorelaxation effects on smaller arteries,and protective effects on endothelial cells,but no effects on the central nervous,respiratory,digestive or endocrine systems at therapeutic dose.Our previous studies demonstrated that iptakalim inhibited the effects of endothelin-1,reduced the intracellular calcium concentration and inhibited the proliferation of pulmonary artery smooth muscle cells.Since iptakalim has been shown safe and effective in both experimental animal models and phase I clinical trials,it can be a potential candidate of HPH in the future.
文摘Increasing evidence, including from our laboratory, has revealed that opening of ATP sensitive potassium channels(K-ATP channels) plays the neuronal protective roles both in vivo and in vitro. Thus K-ATP channel openers(KCOs) have been proposed as potential neuroprotectants. Our previous studies demonstrated that K-ATP channels could regulate glutamate uptake activity in PC12 cells as well as in synaptosomes of rats. Since glutamate transporters(GluTs) of astrocytes play crucial roles in glutamate uptake and KATP channels are also expressed in astrocytes, the present study showed whether and how KATP channels regulated the function of GluTs in primary cultured astrocytes. The results showed that nonselective KCO pinacidil, selective mitochondrial KCO diazoxide, novel, and blood-brain barrier permeable KCO iptakalim could enhance glutamate uptake, except for the sarcolemmal KCO P1075. Moreover pinacidil, diazoxide, and iptakalim reversed the inhibition of glutamate uptake induced by 1-methyl-4-phenylpyridinium(MPP+). These potentiated effects were completely abolished by mitochondrial K-ATP blocker 5-hydroxydecanoate. Furthermore, either diazoxide or iptakalim could inhibit MPP+-induced elevation of reactive oxygen species (ROS) and phosphorylation of protein kinases C(PKC). These findings are the first to demonstrate that activation of K-ATP channel, especially mitochondrial K-ATP channel, improves the function of GluTs in astrocytes due to reducing ROS production and downregulating PKC phosphorylation. Therefore, the present study not only reveals a novel pharmacological profile of KCOs as regulators of GluTs, but also provides a new strategy for neuroprotection.
基金Natural Science Foundation of Shandong Province,No.ZR2015CL016 and No.ZR2011CL012Colleges and Universities of Shandong Province Science and Technology Plan Projects,No.J11LC17
文摘AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)pathway on such an effect.METHODS Male Wistar rats were randomly divided into a control group,a physiological saline(PS)group,a sodium hydrosulfide(Na HS)group,a glibenclamide(Gl)group,Gl plus Na HS group,a pyrrolidine dithiocarbamate(PDTC)group,and a PDTC plus Na HS group.Gastric mucosal injury was induced by RWIS for 3 h in rats,and gastric mucosal damage was analyzed after that.The PS,Na HS(100μmol/kg body weight),Gl(100μmol/kg body weight),Gl(100μmol/kg or 150μmol/kg body weight)plus Na HS(100μmol/kg body weight),PDTC(100μmol/kg body weight),and PDTC(100μmol/kg body weight)plus Na HS(100μmol/kg bodyweight)were respectively injected intravenously before RWIS.RESULTS RWIS induced serious gastric lesions in the rats in the PS pretreatment group.The pretreatment of Na HS(a H2S donor)significantly reduced the damage induced by RWIS.The gastric protective effect of the Na HS during RWIS was attenuated by PDTC,an NF-κB inhibitor,and also by glibenclamide,an ATP-sensitive potassium channel blocker,in a dose-dependent manner.CONCLUSION These results suggest that exogenous H2S plays a protective role against RWIS injury in rats,possibly through modulation of KATP channel opening and the NF-κB dependent pathway.
文摘Objective:To investigate the role of iptakalim,an ATP-sensitive potassium channel opener,in transient cerebral ischemia/reperfusion (I/R) injury and its involved mechanisms.Methods:Intraluminal occlusion of middle cerebral artery (MCAO) in a rat model was used to investigate the effect of iptakalim at different time points.Infarct volume was measured by staining with 2,3,5-triphenyltetrazolium chloride,and immunohistochemistry was used to evaluate the expressions of Bcl-2 and Bax.In vitro,neurovascular unit (NVU) cells,including rat primary cortical neurons,astrocytes,and cerebral microvascular endothelial cells,were cultured and underwent oxygen-glucose deprivation (OGD).The protective effect of iptakalim on NVU cells was investigated by cell viability and injury assessments,which were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and release of lactate dehydrogenase.Caspase-3,Bcl-2 and Bax mRNA expressions were evaluated by real-time polymerase chain reaction (PCR).Results:Administration of iptakalim 0 or 1 h after reperfusion significantly reduced infarct volumes,improved neurological scores,and attenuated brain edema after cerebral I/R injury.Iptakalim treatment (0 h after reperfusion) also reduced caspase-3 expression and increased the ratio of Bcl-2 to Bax by immunohistochemistry.Iptakalim inhibited OGD-induced cell death in cultured neurons and astrocytes,and lactate dehydrogenase release from cerebral microvascular endothelial cells.Iptakalim reduced mRNA expression of caspase-3 and increased the ratio of Bcl-2 to Bax in NVU cells.Conclusions:Iptakalim confers neuroprotection against cerebral I/R injury by protecting NVU cells via inhibiting of apoptosis.
基金supported by grants from National New Drug Research and Development of Key Project(2010ZX09401-307,2008ZX09101-006,2008ZXJ09004-018 and 2009ZX09301-002)
文摘Cumulative evidence suggests that renal vascular endothelial injury play an important role in initiating and extending tubular epithelial injury and contribute to the development of ischemic acute renal failure.Our previous studies have demonstrated that iptakalim's endothelium protection is related to activation of SUR2B/Kir6.1 subtype of ATP sensitive potassium channel(K ATP) in the endothelium.It has been reported that SUR2B/Kir6.1 channels are widely distributed in the tubular epithelium,glomerular mesangium,and the endothelium and the smooth muscle of blood vessels.Herein,we hypothesized that activating renal K ATP channels with iptakalim might have directly neroprotective effects.In this study,glomerular endothelial,mesangial and tubular epithelial cells which are the main cell types to form nephron were exposed to oleic acid(OA) at various concentrations for 24 h.0.25 μl/ml OA could cause cellular damage of glomerular endothelium and mesangium,while 1.25μl/ml OA could lead to the injury of three types of renal cells.It was observed that pretreatment with iptakalim at concentrations of 0.1,1,10 or 100 μmol/L prevented cellular damage of glomerular endothelium and tubular epithelium,whereas iptakalim from 1 to 100 μmol/L prevented the injury of mesangial cells.Our data showed iptakalim significantly increased survived cell rates in a concentration-dependent manner,significantly antagonized by glibenclamide,a K ATP blocker.Iptakalim played a protective role in the main cell types of kidney,which was consistent with natakalim,a highly selective SUR2B/Kir6.1 channel opener.Iptakalim exerted protective effects through activating SUR2B/Kir6.1 channels,suggesting a new strategy for renal injury by its endothelial and renal cell protection.
文摘ATP-sensitive potassium channel(KATP) consists of a 4.4 complex of an inwardly rectifying Kir6.x pore plus a sulfonylurea receptor,which is an ATP-binding cassette transporter.KATP has been indentified in a variety of tissues and recognized as an important drug target.It connects cell metabolism with cell electric activity.KATP has been proposed to play protective roles during heart failure,arrhythmia,myocardial infarction,stress,myocardial ischemia and hypertension.In this review,a summary of KATP is presented with molecular structure,localization,regulation,cardiovascular protective effect and its mechanisms.