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Objective To investigate the effect of sevoflurane preconditioning combined with postconditioning (Spost) on anoxia/reoxygenation (A/R) injury to neonatal rat cardiomyocytes.Methods Primary cultured neonatal rat cardiomyocytes were isolated from SD rats aged 1-3 days and cultured in DMEM liquid culture medium.The cells were seeded in 24-well plates (1 ml/hole),35 mm diameter dishes (5 ml/dish) or in 50 mm culture flasks (8 ml/flask) with a density of 3 × 105/ml and randomly divided into 9 groups (n =24 each):control group (group C),A/R group,Spre group (group S1),Spre + SB203580 group (group S1 + SB),sevoflurane postcon-ditioning (Spost) group (group S2),Spost + SB203580 group(group S2 + SB),Spre + Spost group (group S3),Spre + Spost + SB203580 group (group S3 + SB),and group SB203580 (group SB).The cells were cultured routinely for 160 min in group C and the cells were exposed to 95％ N2-5％ CO2 in an incubator at 37 ℃ for 120 min followed by reoxygenation for 20 min in the other groups.The cells were incubated with 2.5 ％ sevoflurane for 20 min before anoxia in groups S1,S1 + SB,S3 and S3 + SB and in addition SB203580 (specific p38MAPK inhibitor) 5 μmol/L was added simultaneously in groups S1 + SB and S3 + SB.The cells were incubated with 2.5％ sevoflurane for 20 min after beginning of reoxygenation in groups S2,S2-SB,S3 and S3 + SB,and in addition SB203580 5 μmol/L was added simultaneously in groups S2 + SB and S3 + SB.The cells were incubated with SB203580 5 μmol/L for 20 min before anoxia and after beginning of reoxygenation in group SB.The lactate dehydrogenase (LDH) activity,cell survival rate and apoptotic rate were measured at the end of reoxygenation.The levels of phosphor-p38MAPK (p-p38MAPK) was detected at the end of Spre and Spost.Results Compared with group C,the LDH activity and apoptotic rate were significantly increased,while the cell survival rate was significantly decreased in the other groups (P ＜ 0.05).Compared with group A/R,the LDH activity and apoptotic rate were significantly decreased,while the cell survival rate was significantly increased in groups S1,S2 and S3 (P ＜ 0.05).There was no significant difference in the LDH activity,cell survival rate and apoptotic rate between groups S1,S2and S3 (P ＞ 0.05).The myocardial protective effect of Spre or Spost alone or in combination was eliminated by SB203580 (P ＜ 0.05).Spre or Spost alone up-regulated the expression of p-p38MAPK,Spre combined with Spost offered no additional benefit over Spre or Spost alone,and the up-regulative effect was eliminated by SB203580 (P ＜ 0.05).Conclusion Spre combined with Spost produces similar myocardial protective effect with that of either alone and it may because that both Spre and Spost attenuate A/R-induced injury to cardiomyocytes through p38MAPK signaling pathway.
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Objective To investigate the effects of different doses of dexmedetomidine on the minimum alveolar concentration of sevoflurane for blunting responses to skin incision ( MACBAR ) in patients undergoing lower abdominal surgery. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 25-55 yr, weighing 45-75 kg, undergoing electire lower abdominal surgery under general anesthesia, were randomly divided into 4 groups ( n = 15 each): control group (Do group) and 3 dexmedetomidine groups (D1, D2 and D3 groups). The patients were unpremedicated.Dexmedetomidine was not used in group D0. A loading dose of dexmedetomidine 0.1μg/kg was injected iv over 10 min, and then dexmedetomidine was infused at a rate of 0.4, 0.8 and 1.2 μg· kg- 1 · h - 1 for 30 min in groups D1-3 respectively. Anesthesia was induced with inhalation of 8 % sevoflurane. Laryngeal mask airway was inserted when BIS value decreased to 45-55. The patients were mechanically ventilated with inhalation of sevoflurane and a mixture of 50% nitrous oxide and 50% oxygen, and the fresh gas flow was set at 1 L/min. In D0-3 groups, the initial end-tidal concentrations of sevoflurane were 3.0%, 2.5%, 2.0% and 1.5% respectively. The patients' response to skin incision was described as effective if MR or MAP increased by ＜ 15%, or ineffective (MR or MAP increased by ≥ 15%). When the response was effective, the end-tidal concentration of sevoflurane was decreased in the next patient, when ineffective, increased, and the ratio between the two successive concentrations was 0.9.The MRCBAR of sevoflurane was determined by up-and-down method, and 95% confidence interval was calculated.Results The MRCBAR (95% confidence interval) of sevoflurane was 2.85% (2.44%-3.32%), 1.91%(1.61%-2.26%), 1.52% (1.31%-1.77%), and 1.34% (1.15%-1.57%)in D0-3 groups respectively. The MRCBAR of sevoflurane was significantly lower in D1-3 groups than in D0 group, and in D2 and D3 groups than in group D1 (P ＜0.05＝. There was no significant difference in MRCBAR of sevoflurane between D2 and D3 groups (P ＞0.05) .ConclusionContinuous infusion of dexmedetomidine at 0.4, 0.8 and 1.2 μg·kg-1 ·h-1 for 30 min results in a decrease in MACBAR of sevoflurane and enhances the inhibitory effect of sevoflurane on the stress response, and in a dose-dependent manner.
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ObjectiveTo investigate the effect of dexmedetomidine pretreatment on focal cerebral ischemia-reperfusion(I/R) injury in rats.MethodsOne hundred male SD rats weighing 290-310 g were randomly divided into 5 groups(n =20 each):sham operation group(group SH) ; focal cerebral I/R group; focal cerebral I/R + dexmedetomidine 100 μg/kg(group L); focal cerebral I/R+ dexmedetomidine 200 μg/kg (group M) and focal cerebral I/R + dexmedetomidine 400 μg/kg(group H).Focal cerebral I/R was produced by occlusion of middle cerebral artery for 60 min followed by 24 h of reperfusion.Dexmedetomidine 100,200 and 400 μg/kg were injected intraperitoneally 15 min before ischemia in groups L,M and H respectively.While equal volume of normal saline was injected intraperitoneally in group SH.Neurologic function was assessed and scored at 24 h of reperfusion.Then the animals were sacrificed and brains were removed for determination of cerebral infarct volume and microscopic examination.The expression of heat shock protein 70(HSP70) and activity of Na+ -K + -ATPase in ischemic cortex and activity of SOD and concentration of cortisol in plasma were determinationed at 24 h of reperfusion.ResultsCompared with group SH,neurologic deficit scores and plasma concentration of cortisol were significantly increased,activities of SOD and Na+ -K+ -ATPase decreased and expression of HSP70 was up-regulated in groups I/R,L,M and H( P ＜ 0.05).Compared with group I/R,neurologic deficit scores and plasma concentration of cortisol were significantly decreased,activities of SOD and Na+ -K+ -ATPase increased and expression of HSP70 was up-regulated in a dose-dependent manner ( P ＜ 0.05),and the pathological change was reduced in a dose-dependent manner in groups L,M and H.ConclusionDexmedetomidine pretreatment can attenuate focal cerebral I/R injury in a dose-dependent manner through improvement of brain cell energy metabolism and reduction of lipid peroxidation and stress reaction.
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Objective To investigate the effect of sevoflurane preconditioning on CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) expression in the cerebral cortex after focal cerebral ischemiareperfusion (I/R) injury in rats and the mechanism. Methods Thirty-six male SD rats weighing 250-280 g were randomly divided into 3 groups ( n = 12 each) : sham operation group (group S) , focal cerebral I/R group (group I/R) and sevoflurane preconditioning group (group Sevo-pc). The animals were anesthetized with intraperitoneal chloral hydrate 300 mg/kg. In groups I/R and Sevo-pc, focal cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into the right internal carotid artery and advanced cranially until resistance was met. The occlusion was maintained for 1 h followed by 24 h reperfusion. Group Sevo-pc inhaled 2.7% sevoflurane for 1 h before ischemia. Neurological deficits were assessed and scored at the end of 24 h reperfusion and then the rats were decapitated. Their brains were immediately removed. The cerebral infarct size was determined by TTC staining. The CHOP expression in the ischemic cerebral cortex was determined by immunohistochemistry. The number of apoptotic neurons was counted using TUNEL. Results The neurological deficit scores were significantly higher, the cerebral infarct size was significantly larger, and the CHOP expression and the number of apoptotic neurons were significantly higher in groups I/R and Sevo-pc than in group S ( P ＜ 0.01) . The neurological deficit scores were significantly lower, the cerebral infarct size was significantly smaller, and the CHOP expression and the number of apoptosis neurons were significantly lower in group Sevo-pc than in group I/R ( P ＜ 0.05 or 0.01) . Conclusion Sevoflurane preconditioning may protect the brain against focal cerebral I/R injury by down-regulating CHOP expression in the cerebral cortex in rats.
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Objective To investigate the role of mitochondrial ATP-sensitive potassium(mito-KATP)channels in attenuation of ischemia-reperfusion(I/R)injury by lidocaine pretreatment in the isolated rat heart.Methods Adult female Wistar rats weighing 220-250 g were anesthetized with intraperitoneal 3% pentobarbital 35 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%CO2 at 37 ℃.Twenty-four isolated rat hearts with I/R injury were randomly divided into 3 groups(n = 8 each):group I/R,lidocaine group(group L)and lidocaine + glibenclamide group(group LG).After 10 min of equilibration,group C,L and LG received 20 min of perfusion with K-H solution,K-H solution containing lidocaine 2.5 mg/L and K-H solution containing lidocaine 2.5 mg/L + glibenclamide(a blocker of mito-KATP channels)10 μmol/L,respectively,then subjected to 30 min of ischemia followed by 60 min of reperfusion.HR,left ventricular developed pressure(LVDP),+ dp/dtmax and - dp/dtmax were recorded at the end of equilibration(T0)and at 15,30,45 and 60 min of reperfusion(T1-4).Coronary effluent was collected at T0 and T4 for determination of lactate dehydrogenase(LDH)and creatine kinase(CK)activities.Myocardial tissues were obtained from cardiac apex at T4 for determination of Na+ -K+ -ATPase and SOD activities and MDA and Ca2+ contents.Results Compared with group I/R,HR,LVDP,+ dp/dtmax and - dp/dtmax were significantly increased,CK and LHD activities were decreased,Na+ -K+-ATPase and SOD activities were increased,and MDA and Ca2+ contents were decreased in group L(P ＜0.05).Compared with group L,HR,LVDP,+ dp/dtmax and -dp/dtmax were significantly decreased,CK and LHD activities were increased,Na+ -K+ -ATPase and SOD activities were decreased,and MDA and Ca2+ contents were increased in group LG(P＜0.05).Conclusion The mechanism by which lidocaine pretreatment attenuates I/R injury to the isolated rat heart is related to mito-KATP channel opening.
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Objective To investigate the effect of STH-2 cardioplegic solution containing levosimendan on ischemia-reperfusion (I/R) injury in isolated rat hearts. Methods Thirty-two male Wistar rats weighing 250-300 gwere anesthetized with intraperitoneal 3% pentobarbital 30 mg/kg. The hearts were rapidly excised and perfused with oxygenated (95% O2-5% CO2) K-H solution for 30 min in a Langendorff apparatus and then divided into 4groups (n = 8 each) according to the composition of cardioplegic solution: group Ⅰ control (group C) was perfused with STH-2 cardioplegic solution; group Ⅱ , Ⅲ and Ⅳ were peffused with STH-2 cardioplegic solution containing levosimendan 0.03 μmol/L (L1), 0.3 μmol/L (L2) and levosimendan 0.3 μmol/L + glibenclamide 10 μmol/L (L2+ G) respectively. The isolated hearts were first perfused with different cardioplegic solutions for 2 h and then with K-H solution for 30 min. The coronary effluent was collected before ischemia (baseline) and at 10, 20 and 30 min of reperfusion for measurement of creatine kinase (CK) and lactate dehydrogenase (LDH)activities. Myocardial specimens were obtained from apex at 30 min of reperfusion for determination of myocardial ATP and MDA contents and SOD activity. Results Perfusion with STH-2 cardioplegic solution significantly increased CK and LDH activities and MDA content, and significantly decreased SOD activity. Levosinendan 0.03or 0.3 μmol/L significantly attenuated the cardioplegia-induced increase in LDH,CK and SOD activities and MDA content. The protective effects of levosimendan on myocardium against I/R injury were reversed by glibenclamide to some extent. Conclusion Levosimendan can protect myocardium from I/R injury in a dose-dependent manner by opening KATP channel.