Objective To investigate the effects of diazoxide postconditioning on myocardial ischemiareperfusion (I/R) injury in isolated rat hearts. Methods Male SD rats weighing 250-300 g were anesthetized with intraperitoneal pentobarbital 40 mg/kg. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95% O2-5% CO2 at 37 ℃. Sixty-four isolated rat hearts were randomized into 4 groups after 20 min of equilibration (n = 16 each): group control (group C), group I/R, group diazoxide postconditioning (group D) and group mito-KATP channel blocker 5-hydroxydecanoate (5-HD) + diazoxide postconditioning (group 5-HD + D). Group C received continuous perfusion for another 70 min. In group I/R, D and 5-HD + D, the hearts underwent 40 min of iscbemia followed by 30 min of reperfusion. 4 ℃ ST. Thomas cardioplegic solution was administered prior to ischemia in group I/R. Group D received 5 min of perfusion with K-H solution containing 50μ mol/L diazoxide at 5 min of reperfusion. Group 5-HD + D received 5 min of perfusion with K-H solution containing 50 μmol/L 5-HD before reperfusion with diazoxide. Eight hearts were taken at the end of equilibration and reperfusion and the indexes of cardiac function were recorded. Then the mitochondria were extracted for determination of mitochondrial membrane potential (MMP), reactive oxygen species (ROS) release, and respiratory function indexes. Results Compared with group C, MMP was significantly decreased, ROS release was significantly increased, mitochondrial respiratory function and cardiac function declined at the end of reperfusion in the other three groups ( P ＜ 0.05 or 0.01 ). MMP was significantly increased, ROS release was significantly decreased, mitochondrial respiratory function and cardiac function were improved in group D compared with group I/R and 5-HD + D.Conclusion Diazoxide postconditioning can reduce myocardial I/R injury in rats via the opening of mito-KATPchannels and improving the mitochondrial function.

Objective To evaluate the effects of ischemic postconditioning on mitochondrial cardiolipin synthesis during myocardial ischemia-reperfusion (Ⅰ/R) in rats in vitro.Methods Healthy male Sprague-Dawley rats,aged 16-20 weeks,weighing 250-350 g,were used in the study.The animals were anesthetized with intraperitoneal pentobarbital sodium 40 mg/kg and received intraperitoneal heparin 250 U/kg.Their hearts were excised and retrogradely perfused in a Langendorff apparatus.Sixty-four isolated rat hearts were randomly divided into 4 groups (n =16 each) using a random number table:control group (group C),Ⅰ/R group,ischemic postconditioning group (group IPO) and 5-hydroxydecanoate (5-HD) plus ischemic postconditioning group (group 5-HD + IPO).After 20 min of equilibration,the hearts were continuously perfused with K-H solution for 70 min in group C.In Ⅰ/R group,after 20 min of equilibration,the hearts were continuously perfused with 4 ℃ ST.Thomas cardioplegic solution 10 ml/kg,and exposed to 40 min of ischemia followed by reperfusion with oxygenated K-H solution at 37 ℃ for 30 min.In group IPO,after 20 min of equilibration,the hearts were subjected to 6 cycles of 10 s reperfusion followed by 10 s ischemia starting from 40 min of ischemia,and then were reperfused with oxygenated K-H solution at 37 ℃ for 28 min.In group 5-HD + IPO,after 20 min of equilibration,the hearts were perfused with K-H solution containing 100 μmol/L 5-HD (mitochondrial ATP-sensitive potassium channel blocker) for 5 min starting from 40 min of ischemia,and then the other procedures were similar to those previously described in group IPO.At 20 min of equilibration (T1) and 30 min of reperfusion (T2),HR,left ventricular developed pressure (LVDP),left ventricular end-diastolic pressure (LVEDP) and coronary flow (CF) were recorded.The coronary effluent 2 ml was collected for detection of lactic dehydrogenase (LDH) and creatine kinase (CK) activities.The mitochondria were extracted for determination of cardiolipin content.Results HR,LVDP,and CF were significantly lower,LVEDP was higher,and the LDH and CK activities in coronary effluent were higher at T2 than at T1 in the four groups.Compared with group C,HR,LVDP and CF were significantly decreased,LVEDP was increased,and the LDH and CK activities in coronary effluent were increased at T2 in the other three groups.Compared with Ⅰ/R group,HR,LVDP and CF were significantly increased,LVEDP was decreased,and the LDH and CK activities in coronary effluent were decreased at T2 in IPO group.Compared with IPO group,HR,LVDP and CF were significantly decreased,LVEDP was increased,and the LDH and CK activities in coronary effluent were increased at T2 in 5-HD+IPO group.Conclusion The mechanism by which ischemic postconditioning reduces myocardial Ⅰ/R injury is related to opening of mitochondrial ATP sensitive potassium channels and increasing mitochondrial cardiolipin synthesis in rats.

AIM: To investigate the effect of diazoxide (D) postconditioning on Cardiac function and mito-chondrial cardiolipin in isolated rat heart and to explore the protective effect of ATP sensitive potassium channel on diazo-xide postconditioning myocardium.METHODS: The myocardial ischemia/reperfusion injury model in isolated rat hearts was established by Langendorff apparatus.The isolated rat hearts were randomized into 4 groups ( n=8): control group ( control) , myocardial ischemia/reperfusion injury group ( I/R) , diazoxide postconditioning group ( I/R+D) , 5-hydroxy decanoic acid (5-HD) plus diazoxide postconditioning group (I/R+5-HD+D).The hearts in each group were started with 20 min perfusion for equilibration.The hearts in control group perfused for 70 min;The hearts in I/R group was global ischemia for 40 min after ischemia reperfusion at 4℃ST.Thomas cardioplegia, then reperfusion for 30 min;The hearts in I/R+D group were treated with diazoxide (50μmol/L) in K-H perfusion for 5 min after global ischemia for 40 min, then reperfusion for 25 min;The hearts in I/R+5-HD+D group were treated with 5-HD (100μmol/L) in K-H perfusion for 5 min before diazoxide postconditioning, then reperfusion for 20 min.The heart rate, coronary outflow volume, heart func-tion, myocardial enzymes and myocardial mitochondrial cardiolipin at the end of perfusion in each group were determined. RESULTS:Compared with control group and I/R+D group, the heart rate, the concentration of heart phospholipid and the coronary outflow volume were reduced, the heart function was significantly impaired the contents of myocardial enzymes were increased in I/R group.However, no significant difference between I/R group and I/R+5-HD+D group was ob-served.CONCLUSION:The diazoxide postconditioning protects the myocardium by increasing mitochondrial cardiolipin content, reducing the release of myocardial enzymes, improving heart function and reducing myocardial reperfusion injury. The myocardial protective effect of diazoxide is completely blocked by 5-hydroxy decanoic acid.

AIM: To study the protective effect of hyperpolarized cardioplegic arrest on reperfused rat heart performance and to investigate the role of mitochondrial ATP-sensitive K+ channels(mitoKATP) opening in the protection of hyperpolarized cardioplegia against ischemia/reperfusion damage.METHODS: Forty Sprague-Dawley rats were randomized into five groups(n=8 in each group): control group(Con);depolarized arrest group(D);hyperpolarized arrest group(H);depolarized cardioplegia with 5-hydroxydecanoate(5-HD) group(5HD+D);hyperpolarized cardioplegia with 5-HD group(5HD+H).The rat hearts were quickly removed to Langendorff apparatus.The heart perfusion was performed for 20 min with 37 ℃ Krebs-Henseleit buffer balanced with gas mixture(O2∶CO2=95%∶5%) at 5.8 kPa perfusion pressure,then cardial arrest was induced by different cardioplegic solution.Hearts were subjected to ischemia at 37 ℃ for 40 min followed by 30 min reperfusion.(1) The hemodynamics was detected at recovery after 30 min reperfusion.(2) Before ischemia and at the end-reperfusion,tissue was harvested for mitochondrial isolation and ultrastructure was observed by transmission electron microscopy(TEM).(3) Production of reactive oxygen species(ROS) was also determined at different time points.RESULTS:(1) Compared with end-equilibration,30 min reperfusion caused significant differences in left ventricular developed pressure(LADP),left ventricular end-diastolic pressure(LVEDP),double product(DP),heart rate (HR),coronary flow(CF)(P

Objective To investigate the effects of pyrrolidine dithiocarbamate (PDTC) on plasma concentrations of proinflammatory cytokines and myocardial energy metabolism induced by cardiopulmonary bypass (CPB) .Methods Twelve healthy mongrel dogs of both sexes weighing 13.5-17.5 kg were randomly divided into 2 groups (n = 6 in each group) : PDTC group and control group. The animals were anesthetized with intraperitoneal 2.5% pentobarbital sodium 25 mg?kg-1, intubated and mechanically ventilated. In PDTC group PDTC 30 mg?kg-1 was given i.v. after tracheal intubation while in control group normal saline was given instead of PDTC. Aorta was clamped for 60 min and then undamped for 60 min reperfusion during CPB. Blood samples were taken before (baseline), 30 and 60 min after aortic clamping and 30 and 60 min after aortic unclamping for determination of plasma concentrations of TNF-? and IL-1?. Myocardial tissue was obtained before and 60 min after aortic clamping and 60 min after aortic unclamping for determination of myocardial content of adenine nucleotide (ATP, ADP, AMP, TAN, EC) and expression of ICAM-1 protein.Results Plasma TNF-? concentration was increased after aortic cross-clamping as compared to the baseline value before clamping in both groups but the TNF-? concentration was significantly lower in PDTC group than in control group (P

Objective To evaluate the myocardial protective effect of hyperpolarized heart arrest at different temperature during cardiopulmonary bypass CPB.Methods Eighteen dogs were randomly allocated to be infused with St.Thomas solution containing 50?mol/L pinacidil and 5mmol/L K + at 37℃ (group A) or 4℃ (group B), or the standard St.Thomas solution containing K + 16mmol/L at 4℃ (group C) respectively, through aortic root after aortic clamping . The global surgical ischemia lasted 60min with the declamping of 20min .The activities of serum myocardial enzymes, and levels of lipid peroxide and adenine nucleotide of myocardium were measured.Results All paramaters showed to occur serious ischemic and reperfusion damages during the whole procedures in group C, while there were the mild damages in two hyperpolarized groups, especially in group B.Conclusions Myocardial protective effect of hyperpolarized arrest induced with pinacidil, an ATP sensitive potassium channel opener, is superior to that of traditional hyperkalemic cardioplegia , which is much more prominent in hypothermic state.

ObjectiveTo investigate the effect ofpinacidil hyperpolarizaed arrest on p38 mitogen-activited protein kinase (p38MAPK) during myocardial ischemia-reperfusion (I/R) in isolated rat hearts.MethodsFortyeight male SD rats weighting 250-300 g were randomly divided into 6 groups( n =8 each): natural arrest group (group A) ; St.Thomas group (group B) ; pinacidil hyperpolarization arrest group (group C) ;5-hydroxydecanoate (5-HD) group (group D);HMR-1098 group(group E) and 5-HD + HMR-1098 group(group F).Langendorff reperfusion model was established and K-H solution was retrogradely perfused for 15 min.In group A the hearts were arrested naturally afar perfusion was stopped; in group B St.Thomas solution was perfused; in group C pinacidil hyperpolarization solution was perfused; in the other three groups,K-H solution was perfused to isolated rat hearts for 10 min followed by 5 min 5-HD (group D) or HMR-1098(group E) or 5-HD and HMR-1098(group F) perfusion,then hyperpolarization arrest solution was given in each group.Each hearts suffered 60 min ischemia after arrest followed by 30 min K-H solution reperfusion.Coronary flow(CF),HR,left ventricular developed pressure( LVDP),left ventricular systolic pressure(LVSP) and the maximum rate of pressure rise (dp/dtmax) were measured at the end of 15 min K-H solution perfusion and at 20 min of reperfusion.Myocardial phosphatic and nonphosphatic p38MAPK expression was determined by Western blot at 30 min of reperfusion.ResultsCompared with group C,CF,HR,LVDP,LVSP and dp/dtmax were significantly decreased at 20 min of reperfusion and phosphatic p38MAPK expression was down-regulated,non-phosphatic p38MAPK expression was up-regulated at 30 min of reperfusion in groups A,B,D,E and F (P ＜ 0.05).Compared with group E,CF,HR,LVDP,LVSP and dp/ dtmax were significantly decreased at 20 min of reperfusion and phosphatic p38MAPK expression was down-regulated,non-phosphatic p38MAPK expression was up-regulated at 30 min of reperfusion in groups D and F ( P ＜0.05).ConclusionHyperpolarized arrest induced by pinacidil can improve cardiac function following myocardial I/R injury by up-regulating phosphatic p38MAPK expression,down-regulating non-phosphatic p38 MAPK expression and mitochondrial potassium channel is more important than membranous one during the regulation of phosphatic p38MAPK expression.

Objective To evaluate the role of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in cardio-protection by ischemic or pinacidil postconditioning ( IP,PP) against ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Fifty-six male SD rats of both sexes weighing 200-250 g were anesthetized with intraperitoneal amobarbital sodium.The isolated rat hearts were perfused in a Langendorff apparatus with Krebs-Hensleit buffer (K-H).Fifty-six isolated rat hearts with I/R injury were randomly divided into 7 groups ( n =8 each):normal control group (group C) ; group I/R; group IP and group PP1-4 postconditioning with 4 different concentrations of pinacidil.After 20 min of equilibration,the perfusion was suspended for 40 min (global ischemia) followed by 60 min of reperfusion in group I/R.In group IP after 40 min of global ischemia,the isolated hearts underwent 6 cycles of 10 s reperfusion and 10 s ischemia followed by 58 min of reperfusion.In group PP1-4 at the end of 40 min of global ischemia,the isolated hearts were perfused with K-H containing pinacidil 5,10,30 and 50μmol/L for 5 min respectively followed by 55 min reperfusion with regular K-H.Left ventricular developed pressure (LVDP) and LVEDP were measured immediately before global ischemia and at the end of 60 min reperfusion.Myocardial specimens were obtained at the end of reperfusion for detection of Nrf2,quinopeoxidoreductase (NQO1),HO-1 and SOD1 mRNA (by RT-PCR) and protein (by Western blot) expression.Results I/R significantly up-regulated Nrf2,NQO1,HO-1 and SODI mRNA and protein expression,decreased LVDP and increased LVEDP in group I/R as compared with group C.IP and 30,50 μmol/L pinacidil postconditioning further significantly increased Nrf2,NQO1,HO-1 and SOD1 mRNA and protein expression and IP,5,10,30,50 μmol/L pinacidil postconditioning significantly increased LVDP and decreased LVEDP as compared with group I/R.Conclusion Ischemic or pinacidil postconditioning can attenuate I/R injury by activating Nrf2-ARE pathway in isolated rat hearts.

Astrocytes were traditionally be deemed as supportive cells in the central nervous system.However,recent researches proved astrocytes exerted more important neurophysiological functions,such as regulation of synaptic transmission and integration of neural information.This paper summarized the researches on the functions of astrocytes related to synaptic transmission and analyzed the new features of astrocyte excitability,the communication between neurons and astrocytes and the effect of general anesthetics on astrocytes.Based on these new findings,this paper also suggested the underlying relationship between astrocytes and the loss of consciousness during general anesthesia.

Objective To investigate the effect of heart preservation solution containing pinacidil on mitochondrial function in isolated rat hearts. Methods One hundred and twenty pathogen-free SD rats of both sexes weighing 250-350 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 65 mg/kg. Their hearts were immediately removed and perfused in a Langendorff apparatus. Left ventricular enddiastolic pressure was measured from a fluid-filled latex balloon inserted in the left ventricle. The isolated hearts were randomized into 5 groups (n = 24 each):group Ⅰ was perfused with cardioplegic solution HTK; group Ⅱ with HTK containing pinacidil (a non-specific sarcKATP and mitoKATP channel opener) 0.5 mmol/L; group Ⅲ with HTK containing pinacidil 0.5 mmol/L + 5-HD (a selective mitoKATP channel blocker) 100 μmol/L; group Ⅳ with HTK containing pinacidil 0.5 mmol/L + HMR-1098 100 μmol/L (a selective sarcKATP channel blocker) and group Ⅴ with HTK containing pinacidil 0.5 mmol/L + 5-HD 100 μmol/L + HMR-1098 100μmol/L. The isolated hearts were perfused with simple HTK or HTK containing pinacidil or pinacidil + 5-HD and/or HMR 20 ml/kg at 10 ml/min and then removed from Langendorff apparatus and dipped into the same HTK solution for 8 h at 4 ℃followed by 60 min reperfusion. The respiratory function of mitochondria (respiratory control rate (RCR), the rate of oxygen consumption in state 3/state 4 and P/O) was measured at the end of equilibration (T1) after 8 hpreservation (T2) and at the end of 60 min reperfusion (T3). The CK-MB and LDH activities and cTnI expression in myocardium was detected at T1 and T3. The ultrastructure of myocardium was examined at T3. Results Perfusion suspension-reperfusion (PS/R) significantly decreased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and increased myocardial cTnI concentration and CK-MB and LDH activities at T3 compared with baseline at T1 in group Ⅰ. Pinacidil significantly increased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and decreased myocardial cTnI concentration and CK-MB and LDH activities in group Ⅱ as compared with group Ⅰ-indicative of protective effect of pinacidil on mitochondria against PS/R injury. The protective effect of pinacidil against PS/R injury was attenuated by 5-HD and/or HMR1098. The myocardial damage was slightest in group Ⅱ . Conclusion Both sarcolemmal and mitochondrial KATPchannel are involved in the protective effect of pinacidil against PS/R-induced myocardial damage during heart preservation.