Objective To determine the protection effects and mechanisms for immature myocardium with limbs ischemic preconditioning (LIP). Methods Using the Langendorff perfusion apparatus to perfuse isolated hearts, we randomly divided 30 Japanese longeared white rabbits into 5 groups, each having 6 rabbits. For the I/R group，after the perfusion model was established, the isolated hearts underwent 15 min of perfusion with KH solution before working for another 15 min . Then perfusion was stopped to cause ischemia for 45 min before reperfusion for 15 min and working for another 30 min . For E1 group, the model was established by 3×LIP (double limbs obstructed for 5 min followed by 5 min reperfusion for 3 times) and then procedures of the I/R group were carried out. For E2 group, before procedures of the E1group were done, superoxide dismutase (SOD) was injected till LIP was completed. For E3 group, intravenous protein kinase C (PKC) polymyxin (PMB) was injected for 10 minutes before E1 procedures were repeated. For E4 group, intravenous mitochondrial ATPsensitive K+ channels (mitoKATP) blocker 5-hydroxydecanoate was injected for 10 min before E1 procedures were carried out. The left ventricular function recovery, myocardial water content (MWC), creatine kinase (CK) and lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA) and ATP content, SOD activity and superoxygen negative ion (O2 ·-) content were tested. Results Left ventricular recovery in E1 group was better than other groups (Plt;0.05). ATP content and SOD activity in E1 group were also better than all other groups (Plt;0.05). MWC in E1 group was lower than other groups (Plt;0.05). MDA content, CK and LDH leakage in E1 group were also lower than other groups (Plt;0.05). There was no significant difference of the above indications among I/R,E2,E3 and E4 groups, while the difference of O2·- content in E1,E3 and E4 groups before and after preconditioning was significant. Conclusion LIP has obvious protective effects for immature myocardium and the mechanisms are probably through PKC stimulation and opening of mitoKATP.
Abstract： Objective To investigate the mechanism of protein kinase C(PKC) in immature myocardial ischemic preconditioning in order to further its clinical applicability. Methods Langendorff perfusion heart models of 24 rabbits were set up and they were randomly divided into 4 groups: ischemic reperfusion group (I/R group), myocardial ischemic preconditioning group (MIP group), chelerythrine group (CLT group) and protein kinase C group (PKC group). The emodynamics, biochemistry and myocardial ultrastructure were observed. Results The heart function recovery and myocardial water content in the MIP and the PKC groups were better than those of the I/R and the CLT groups (Plt;0.01). The adenosine triphosphate (ATP) content, superoxide dismutase activity, mitochondrial Ca2+-ATPase activity and synthesizing ATP activity of mitochondria in the MIP and the PKC groups were significantly higher than those of the I/R and the CLT groups (Plt;0.01). The dehydrogenase and creatine kinase leakage, malondialdehyde content, myocardial cell Ca2+ content and mitochondrial Ca2+ content in the MIP and the PKC groups were significantly lower than those of the I/R and the CLT groups (Plt;0.01). The myocardial ultrastructure injuries in the MIP and the PKC groups were less than that of the I/R and the CLT groups. Conclusion Myocardial ischemic preconditioning plays an important role in protecting immature myocardium, which is probably realized by the activation of PKC.
Objective To investigate the role of mitochondrial adenosine triphosphatesensitive potassium channel(mitoKATP) in immature myocardial ischemic preconditioning, and to provide evidence for immature myocardial protection. Methods Langendorff isolated heart infused model was used in the experiment. Twentyfour rabbits (aged from 14 to 21 days) were randomly divided into 4 groups：ischemiareperfusion group（I/R group）, myocardial ischemic preconditioning group(E1 group), 5hydroxydecanoate(5-HD) group (E2 group) and Diazoxide (Diaz) group(E3 group). Hemodynamics recovery rate, myocardial water content(MWC), the leakage rates of serum creatine kinase and lactate dehydrogenase, adenosine triphosphate content, superoxide dismutase activity, malondialdehyde content, myocardial cell Ca2+ content and myocardial mitochondrial Ca2+ content, myocardial mitochondrial Ca2+-ATPase activity, the adenosine triphosphate(ATP) synthesizing ability of myocardial mitochondria were tested, and myocardial ultrastructure was observed via electron microscopy. Results The hemodynamics recovery rate, myocardial water content(P＜0.05), adenosine triphosphate content, superoxide dismutase activity, myocardial mitochondrial Ca2+-adenosine triphosphyatase(ATPase) activity and the ATP synthesizing ability of myocardial mitochondria of the rabbits in E1 and E3 group were significantly better than that in I/R group and E2 group(P＜0.05). Malondialdehyde content, the leakage rates of serum creatine kinase and lactate dehydrogenase, myocardial cell Ca2+ content and myocardial mitochondrial Ca2+ content of the rabbits in E1 group and E3 group were significantly lower than that in I/R group and E2 group (P＜0.05). The myocardial ultrastructure injury in E1 and E3 group were significantly reduced compared with that in I/R and E2 group. Conclusion Myocardial ischemic preconditioning has significant protective effects on immature myocardium. Its mechanism may be related to the activation of mitoKATP.
Objective To investigate the immature myocardial protection effects with renal ischemic preconditioning. Methods 18 neonatal rabbits were randomly divided into three groups. Ischemic/reperfusion(I/R) group underwent 45 min ischemia followed with 45 min reperfusion after Langendorff model performed. Cardiac ischemic preconditioning(CIP) group underwent 45 min ischemia followed with 45 min reperfusion after 5 min ischemia and then 5 min reperfusion for two times. Renal ischemic preconditioning(RIP) group underwent 45 min ischemia followed with 45 min reperfusion after renal artery obstruction for 5 min and 5 min reperfusion for three times.The left ventricular function recovery,myocardial water content(MWC), lactate dehydrogenase (LDH) and creatine kinase(CK) leakage, malondialdehyde(MDA) content,adenosine triphosphate(ATP) content, superoxide dismutase(SOD) activity, myocardial cell Ca2+ ［Ca2+］c content,mitochondrial Ca 2+ content ［Ca2+］m,synthesizing ATP activity of mitochondria ［ATP］m and Ca2+ATPase activity of mitochondria ［Ca2+ATPase］m were tested. Results The recovery of postischemic heart function in RIP group and CIP group were higher than that I/R group(Plt;0.01). There were no significant difference of HR, AF in three groups （Pgt;0.05）. There were significant difference of CF，CO，LVSP and LVEDP in RIP group and CIP group than those I/R group（Plt;0.01）. There were significant difference of MWC, CK, LDH, ATP content, MDA, SOD activity, ［Ca2+］c content, ［Ca2+ATPase］m， ［Ca2+］m and ［ATP］m in RIP group than those I/R group（Plt;0.01). There were no significant difference between RIP group and CIP group upon every index (Pgt;0.05). Conclusion RIP has the same cardioprotection to immature myocardium as ischemic preconditioning.
Objective To investigate the protective effects of metallothionein （MT） for immature myocardium and myocardial interstitium.Methods Twenty-four rabbits （aged 14-21 days） were divided into 4 groups with random number table, 6 each group. Distilled water was injected intraperitoneally in control group and 3.6% ZnSO4 （1.5 ml/kg） was injected intraperitoneally in group 1, group 2 and group 3. Control group, group 1, group 2 and group 3 isolated working rabbit heart model were used in 24h, 12h, 24h and 48h after intraperitoneally respectively. The MT content, recovery of hemodynamics, biochemistry and myocardial ultrastructure were tested. Results The MT content, hemodynamics recovery, adenosine triphosphate （ATP） content, superoxide dismutase activity, Ca2＋-ATPase activity, synthesizing ATP activity of mitochondria, hydroxyproline in group 2 and group 3 were higher than those in control group and group 1 （P 〈 0. 01）. The myocardial watery content, creatine kinase and dehydrogenase leakage, malondialdehyde content, endothelin, mitochondrial Ca2＋ content in group 2 and group 3 were lower than those in control group and group 1（P〈0.01）. The myocardial ultrastructure injuries were less in group 2 and group 3 than that in control group and group l. Conclusion This study demonstrates that myocardial MT prolonged expression can be induced by ZnSO4 and the ischemia-reperfusion injury of immature myocardium and myocardial interstitium could be reduced by MT.
Objective To investigate the protection effects of different pH N 2 hydroxyethyl piperazine N′ 2 ethanesulfonic acid (HEPES) Krebs Henseleit (KH) reperfusate solutions on immature myocardium and myocardial interstitium. Methods Isolated perfused Langendorff model from immature rabbit hearts were performed. Twenty four rabbits were divided into control group, ischemia reperfusion group and acidic reperfusate group. The left ventricular function recovery, myocardial water content(MWC), lactate dehydrogenase (LDH) and creatine kinase (CK) leakage, malondialdehyde(MDA) and adenosine triphosphate (ATP) content, superoxide dismutase(SOD) activity , hydroxyproline(HP) and endothelin(ET) content were tested. Results The left ventricular functional recovery, ATP content, SOD activity and HP content in acidic reperfusate group were higher than those of ischemia reperfusion group( P lt;0.05).MWC, MDA content, CK and LDH leakage, and ET content in acidic reperfusate group were lower than those of ischemia reperfusion group( P lt;0.05). Conclusion These results suggest pH paradox might be one of important mechanisms for immature myocardium and myocardial interstitium ischemia reperfusion injury,and acidic perfusate, at the beginning of reperfusion ,might attenuate pH paradox and ameliorate functional recovery in isolated perfused immature rabbit hearts.
Objective To improve the myocardial protection result, observe the effects of 11,12 epoxyeicosatrienoic acid (11,12 EET) on reperfusion arrhythmias in the isolated perfused immature rabbit hearts, which underwent long term preservation. Methods Sixteen isolated rabbit hearts were randomly assigned to two groups, 8 rabbits each group. Control group: treated with St.Thomas Ⅱ solution, experimental group: treated with St.Thomas Ⅱ solution plus 11,12 EET. By means of the Langendorff technique, these isolated rabbit hearts were arrested and stored for 16 hours with 4℃ hypothermia, and underwent 30 minutes of reperfusion(37℃). The mean times until the cessation of both electrical and mechanical activity were measured after infusion of cardioplegia. The heart rate (HR), coronary flow (CF), myocardial water content (MWC), value of creatine kinase (CK) and lactic dehydrogenase (LDH), myocardial calcium content and the arrhythmias score (AS) during the period and at the endpoint of the reperfusion were observed. Results The times until electrical and mechanical activity arrest in the experimental group were significantly shorter than those in control group ; HR, CF, MWC, CK, LDH, myocardial calcium content and AS were significantly better than those in control group. Conclusions These data suggest that 11,12 EET added to the cardioplegic solution of St.Thomas Ⅱ has lower incidence rate of reperfusion arrhythmias.
ObjectiveTo compare the myocardial protective effect of HTK solution and St.ThomasⅡ(STH) solution in immature rabbit myocardium at different cardiac arrest time. MethodsAccording to cardioplegia and cardiac arrest time, 32 immature New Zealand white rabbits (aged 2-3 weeks) were randomly divided into four groups. A group SO (8 rabbits) underwent 1 hour cardiac arrest with STH solution, a group ST (8 rabbits) underwent 2 hours cardiac arrest with STH solution, a group HO (8 rabbits) underwent 1 hour cardiac arrest with HTK solution, a group Ht (8 rabbits) underwent 2 hours cardiac arrest with HTK solution. Compare the myocardial protective effect of HTK and STH solution in immature myocardium at different cardiac arrest time. ResultsThe Langendorff models were successfully established in 30 cases (8 cases in the group SO and HO, 7 cases in the group ST and HT). There were no statistical differences in hemodynamics and myocardial enzyme (CK-MB, LDH) (P > 0.05), but HTK solution reduced the activity of nitric oxide synthase (NOS) and content of malonaldehyde (MDA) and NO, maintained high activity of superoxide dismutase (SOD) and Ca2+-ATPase (P < 0.05), performed more effective myocardial protection for immature myocardium. ConclusionHTK solution has more effective myocardial protection for immature myocardium than STH solution does, but STH solution still has good outcomes within short cardiac arrest time (1h).