七氟烷預處理和后處理對離體大鼠心臟再灌注心律失常和心肌細胞電生理的影響
發(fā)布時間：2018-08-11 11:40
【摘要】：目的： 1、觀察七氟烷預處理(Sevoflurane preconditioning, SpreC)和七氟烷后處理(Sevoflurane postconditioning, SpostC)對再灌注心律失常的影響,探索其相關機制； 2、觀察SpreC和SpostC對離體大鼠心肌細胞動作電位、L型鈣通道電流(L-type calcium current, ICa, L)、快鈉通道電流(Fast sodium current,INa)和瞬時外向鉀通道電流(Transient outward potassium current, Ito)電生理特性的影響及相關機制； 3、比較SpreC和SpostC對再灌注心律失常和離子通道電生理作用的異同。 方法： 1、取40只SD大鼠心臟建立Langendorff灌注模型, KH液平衡15min后隨機分入以下4組,每組10只：(1)時間對照組(Time Control,TC),平衡期后繼續(xù)灌注KH液75min；(2)缺血再灌注組(Ischemia/reperfusion,I/R),平衡期后灌注KH液20min,隨后停灌25min復灌30min制作全心缺血再灌注損傷模型；(3)七氟烷預處理組(SpreC),在全心停灌前以3%七氟烷預處理15min,普通KH液洗脫5min,隨后停灌25min,復灌30min；(4)七氟烷后處理組(SpostC),平衡期后灌注普通KH液20min,停灌25min,復灌30min,在復灌之初使用3%七氟烷后處理15min。比較各組血流動力學、冠脈流出液肌鈣蛋白I、心肌梗死面積、再灌注心律失常及細胞內鈣離子和活性氧水平。 2、取SD大鼠心臟制備Langendorff灌注模型,分組及缺血再灌注方案同第一部分,再灌注結束后分離各組心室肌外膜細胞進行電生理實驗。使用標準的全細胞膜片鉗技術記錄各組心室肌細胞的動作電位振幅、靜息膜電位和動作電位時程；記錄各組心室肌細胞的ICa,L、INa、Ito,分析其電流-電壓曲線、穩(wěn)態(tài)激活曲線、穩(wěn)態(tài)失活曲線和穩(wěn)態(tài)失活后恢復曲線的變化,以了解SpreC和SpostC對大鼠心室肌細胞電生理特性的影響。 結果： 1、血流動力學、心肌損傷和再灌注心律失常相關結果。與I／R組相比,SpreC組和SpostC組能增大再灌注心臟的左室發(fā)展壓、左室最大收縮／舒張速率和心率,降低左室舒張末期壓力,降低冠脈流出液肌鈣蛋白I水平,減少心梗面積(P0.05)。SpreC和SpostC減少離體心臟再灌注期間VPB的發(fā)生個數(shù)、減少VF的發(fā)生率、縮短VT和VF的發(fā)作時程、降低再灌注心律失常評分(P0.05)。SpreC和SpostC降低再灌注后心肌細胞內鈣離子水平和活性氧水平(P0.05)。SpreC和SpostC在上述指標的差異未見統(tǒng)計學意義(P0.05)。 2、心肌細胞電生理相關結果。(1)動作電位：與TC組相比,I／R組的心肌細胞動作電位振幅減小、靜息膜電位升高、動作電位時程縮短(P0.05)。與I／R組相比,SpreC和SpostC能增大動作電位振幅、降低靜息膜電位、延長動作電位時程(P0.05)。(2)L型鈣通道：與TC組相比,I／R組ICa，L的峰值密度降低,半失活電壓減小,失活曲線左移,恢復時間常數(shù)增大(P0.05)。與I／R組相比,SpreC和SpostC能增加ICa,L的峰值密度,增加ICa,L失活曲線的半失活電壓,右移失活曲線,降低ICa，L恢復曲線的時間常數(shù)(P0.05)。(3)快鈉通道：與TC組相比,I／R組INa的峰值密度降低,穩(wěn)態(tài)失活曲線的斜率因子k降低(P0.05)。與I／R組相比,SpreC和SpostC能增加INa的峰值密度,增加失活曲線的斜率因子k(P0.05)。(4)瞬時外向鉀通道：與TC組相比,I／R組Ito的峰值密度降低,半激活電壓和半失活電壓增大,穩(wěn)態(tài)激活曲線和穩(wěn)態(tài)失活曲線右移(P0.05)。與I／R組比較,SpreC和SpostC能增加Ito的峰值密度,減小半失活電壓,左移失活曲線(P0.05)。SpreC和SpostC關于動作電位、ICa,L、INa和Ito相關指標之間的差異無統(tǒng)計學意義(P0.05)。 結論： 1、SpreC和SpostC能改善缺血再灌注心臟的血流動力學,減輕心肌損傷,減少心肌梗死面積,對離體大鼠心臟發(fā)揮保護效應。SpreC和SpostC能改善離體大鼠再灌注心律失常的發(fā)生及發(fā)作。上述作用可能與SpreC和SpostC降低心肌細胞內鈣離子和氧自由基水平有關。 2.(1) SpreC和SpostC能減小缺血再灌注損傷對動作電位參數(shù)的影響。(2)SpreC和SpostC能增加ICa,L峰值密度,加快失活.,減慢恢復,緩解缺血再灌注損傷造成的變化,提示SpreC和SpostC有利于延長再灌注損傷后心肌細胞的動作電位時程,緩解心肌復極離散度的增大。此外,SpreC和SpostC對L型鈣通道的調控有利于維持鈣穩(wěn)態(tài),緩解再灌注損傷后鈣超載。(3)SpreC和SpostC能增加再灌注損傷后INa的峰值密度,提示SpreC和SpostC能增加再灌注損傷心肌細胞動作電位的0相上升速率,加快沖動傳導,對興奮性折返性心律失常有一定的預防作用。(4)SpreC和SpostC能增加再灌注損傷心肌細胞Ito的峰值密度,降低半失活電壓,加快失活過程,能減少缺血再灌注損傷對動作電位時程的影響,減少復極離散度,減少Ito介導的2相折返和后除極的發(fā)生率,從而緩解再灌注心律失常的發(fā)生及發(fā)作。 3、SpreC和SpostC對再灌注心臟的血流動力學、心肌酶釋放水平、心梗面積、再灌注心律失常等方面的影響類似。SpreC和SpostC對動作電位、ICa，，L、INa和Ito的電生理特性的影響類似。
[Abstract]:Objective:
1. To observe the effects of sevoflurane preconditioning (SpreC) and sevoflurane postconditioning (SpostC) on reperfusion arrhythmia and explore its related mechanism.
2. To observe the effects of SpreC and Post C on action potential, L-type calcium current (ICa, L), Fast sodium current (INa) and transient outward potassium channel current (Ito) in isolated rat cardiomyocytes and the related mechanisms.
3, we compared the electrophysiological effects of SpreC and SpostC on reperfusion arrhythmias and ion channels.
Method:
1. 40 SD rats were randomly divided into four groups after 15 minutes of KH fluid balance: (1) time control group (TC), after the balance period, KH fluid was continuously perfused for 75 minutes; (2) ischemia / reperfusion group (I / R), after the balance period, KH fluid was perfused for 20 minutes, and then stopped for 25 minutes to reperfusion for 30 minutes. Cardiac ischemia-reperfusion injury model; (3) Sevoflurane preconditioning group (SpreC), before cardiac arrest with 3% sevoflurane preconditioning for 15 minutes, elution of ordinary KH solution for 5 minutes, then stop perfusion for 25 minutes, reperfusion for 30 minutes; (4) Sevoflurane postconditioning group (SpostC), after equilibrium perfusion of ordinary KH solution for 20 minutes, stop perfusion for 25 minutes, reperfusion for 30 minutes, at perfusion at the beginning of reperfusion with 3% sevoflurane postconditioning. 15 min. The hemodynamics, cardiac troponin I, myocardial infarction area, reperfusion arrhythmia, intracellular calcium ion and reactive oxygen species were compared.
2. Langendorff perfusion model was established in SD rat hearts. The ventricular epicardial cells were isolated from each group after reperfusion and electrophysiological experiments were performed. The amplitude of action potential, resting membrane potential and action potential duration of ventricular myocytes were recorded by standard whole cell patch clamp technique. ICa, L, INa, Ito of ventricular myocytes were recorded, and their current-voltage curves, steady-state activation curves, steady-state inactivation curves and recovery curves after steady-state inactivation were analyzed to understand the effects of SpreC and Post-C on the electrophysiological characteristics of ventricular myocytes in rats.
Result:
1. Hemodynamics, myocardial injury and reperfusion arrhythmias. Compared with I/R group, Spre C and Spost C groups increased left ventricular development pressure, maximal left ventricular systolic/diastolic rate and heart rate, decreased left ventricular end-diastolic pressure, decreased coronary effluent troponin I levels, and decreased myocardial infarction area (P 0.05). OstC decreased the number of VPB, the incidence of VF, the duration of VT and VF, and the reperfusion arrhythmia score (P 0.05). SpreC and postC decreased the levels of intracellular calcium and reactive oxygen species (P 0.05). There was no significant difference between SpreC and postC (P 0.0). 5).
2. Relevant electrophysiological results of myocardial cells. (1) Action potential: Compared with TC group, the amplitude of action potential decreased, resting membrane potential increased and action potential duration shortened in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase action potential amplitude, decrease resting membrane potential and prolong action potential duration (P 0.05). Track: Compared with TC group, the peak density of ICa and L decreased, the half-inactivation voltage decreased, the inactivation curve shifted to the left, and the recovery time constant increased in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase the peak density of ICa and L, increase the half-inactivation voltage of ICa and L inactivation curves, and decrease the time constant of ICa and L recovery curves (P 0.05). 3) Fast sodium channel: Compared with TC group, the peak density of INa and the slope factor K of steady-state inactivation curve decreased in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase the peak density of INa and the slope factor K of inactivation curve (P 0.05). (4) Instantaneous outward potassium channel: Compared with TC group, the peak density of Ito in I/R group was decreased and the semi-activated potassium channel was increased. Compared with the I/R group, SpreC and Post C could increase the peak density of Ito, decrease the half-inactivation voltage, and shift the inactivation curve to the left (P 0.05). There was no significant difference between SpreC and Post C on action potential, ICa, L, INa and Ito (P 0.05).
Conclusion:
1. SpreC and postC can improve the hemodynamics of ischemia-reperfusion heart, alleviate myocardial injury, reduce the size of myocardial infarction, and protect isolated rat heart. SpreC and postC can improve the occurrence and attack of reperfusion arrhythmia in isolated rats. These effects may be related to SpreC and postC reducing intracellular calcium ions and oxygen spontaneity. Related to base level.
2. (1) SpreC and postC can reduce the effect of ischemia-reperfusion injury on action potential parameters. (2) SpreC and postC can increase the peak density of ICa and L, accelerate inactivation, slow recovery, and alleviate the changes caused by ischemia-reperfusion injury, suggesting that SpreC and postC can prolong the action potential duration of myocardial cells after reperfusion injury and alleviate myocardial recovery. In addition, the regulation of L-type calcium channel by SpreC and Post C is beneficial to maintain calcium homeostasis and relieve calcium overload after reperfusion injury. (3) SpreC and Post C can increase the peak density of INa after reperfusion injury, suggesting that SpreC and Post C can increase the phase-0 rise rate of action potential and accelerate impulse conduction in myocardial cells after reperfusion injury. (4) SpreC and postC can increase the peak density of Ito, decrease the half-inactivation voltage, accelerate the inactivation process, reduce the effect of ischemia-reperfusion injury on action potential duration, reduce repolarization dispersion, and reduce Ito-mediated phase 2 reentry and depolarization. Thus, the incidence and incidence of reperfusion arrhythmia can be alleviated.
3. The effects of SpreC and Post C on hemodynamics, myocardial enzyme release, myocardial infarction area and reperfusion arrhythmia were similar. The effects of SpreC and Post C on action potential, ICa, L, INa and Ito were similar.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R614;R-332
本文編號：2176895
[Abstract]:Objective:
1. To observe the effects of sevoflurane preconditioning (SpreC) and sevoflurane postconditioning (SpostC) on reperfusion arrhythmia and explore its related mechanism.
2. To observe the effects of SpreC and Post C on action potential, L-type calcium current (ICa, L), Fast sodium current (INa) and transient outward potassium channel current (Ito) in isolated rat cardiomyocytes and the related mechanisms.
3, we compared the electrophysiological effects of SpreC and SpostC on reperfusion arrhythmias and ion channels.
Method:
1. 40 SD rats were randomly divided into four groups after 15 minutes of KH fluid balance: (1) time control group (TC), after the balance period, KH fluid was continuously perfused for 75 minutes; (2) ischemia / reperfusion group (I / R), after the balance period, KH fluid was perfused for 20 minutes, and then stopped for 25 minutes to reperfusion for 30 minutes. Cardiac ischemia-reperfusion injury model; (3) Sevoflurane preconditioning group (SpreC), before cardiac arrest with 3% sevoflurane preconditioning for 15 minutes, elution of ordinary KH solution for 5 minutes, then stop perfusion for 25 minutes, reperfusion for 30 minutes; (4) Sevoflurane postconditioning group (SpostC), after equilibrium perfusion of ordinary KH solution for 20 minutes, stop perfusion for 25 minutes, reperfusion for 30 minutes, at perfusion at the beginning of reperfusion with 3% sevoflurane postconditioning. 15 min. The hemodynamics, cardiac troponin I, myocardial infarction area, reperfusion arrhythmia, intracellular calcium ion and reactive oxygen species were compared.
2. Langendorff perfusion model was established in SD rat hearts. The ventricular epicardial cells were isolated from each group after reperfusion and electrophysiological experiments were performed. The amplitude of action potential, resting membrane potential and action potential duration of ventricular myocytes were recorded by standard whole cell patch clamp technique. ICa, L, INa, Ito of ventricular myocytes were recorded, and their current-voltage curves, steady-state activation curves, steady-state inactivation curves and recovery curves after steady-state inactivation were analyzed to understand the effects of SpreC and Post-C on the electrophysiological characteristics of ventricular myocytes in rats.
Result:
1. Hemodynamics, myocardial injury and reperfusion arrhythmias. Compared with I/R group, Spre C and Spost C groups increased left ventricular development pressure, maximal left ventricular systolic/diastolic rate and heart rate, decreased left ventricular end-diastolic pressure, decreased coronary effluent troponin I levels, and decreased myocardial infarction area (P 0.05). OstC decreased the number of VPB, the incidence of VF, the duration of VT and VF, and the reperfusion arrhythmia score (P 0.05). SpreC and postC decreased the levels of intracellular calcium and reactive oxygen species (P 0.05). There was no significant difference between SpreC and postC (P 0.0). 5).
2. Relevant electrophysiological results of myocardial cells. (1) Action potential: Compared with TC group, the amplitude of action potential decreased, resting membrane potential increased and action potential duration shortened in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase action potential amplitude, decrease resting membrane potential and prolong action potential duration (P 0.05). Track: Compared with TC group, the peak density of ICa and L decreased, the half-inactivation voltage decreased, the inactivation curve shifted to the left, and the recovery time constant increased in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase the peak density of ICa and L, increase the half-inactivation voltage of ICa and L inactivation curves, and decrease the time constant of ICa and L recovery curves (P 0.05). 3) Fast sodium channel: Compared with TC group, the peak density of INa and the slope factor K of steady-state inactivation curve decreased in I/R group (P 0.05). Compared with I/R group, SpreC and Post C could increase the peak density of INa and the slope factor K of inactivation curve (P 0.05). (4) Instantaneous outward potassium channel: Compared with TC group, the peak density of Ito in I/R group was decreased and the semi-activated potassium channel was increased. Compared with the I/R group, SpreC and Post C could increase the peak density of Ito, decrease the half-inactivation voltage, and shift the inactivation curve to the left (P 0.05). There was no significant difference between SpreC and Post C on action potential, ICa, L, INa and Ito (P 0.05).
Conclusion:
1. SpreC and postC can improve the hemodynamics of ischemia-reperfusion heart, alleviate myocardial injury, reduce the size of myocardial infarction, and protect isolated rat heart. SpreC and postC can improve the occurrence and attack of reperfusion arrhythmia in isolated rats. These effects may be related to SpreC and postC reducing intracellular calcium ions and oxygen spontaneity. Related to base level.
2. (1) SpreC and postC can reduce the effect of ischemia-reperfusion injury on action potential parameters. (2) SpreC and postC can increase the peak density of ICa and L, accelerate inactivation, slow recovery, and alleviate the changes caused by ischemia-reperfusion injury, suggesting that SpreC and postC can prolong the action potential duration of myocardial cells after reperfusion injury and alleviate myocardial recovery. In addition, the regulation of L-type calcium channel by SpreC and Post C is beneficial to maintain calcium homeostasis and relieve calcium overload after reperfusion injury. (3) SpreC and Post C can increase the peak density of INa after reperfusion injury, suggesting that SpreC and Post C can increase the phase-0 rise rate of action potential and accelerate impulse conduction in myocardial cells after reperfusion injury. (4) SpreC and postC can increase the peak density of Ito, decrease the half-inactivation voltage, accelerate the inactivation process, reduce the effect of ischemia-reperfusion injury on action potential duration, reduce repolarization dispersion, and reduce Ito-mediated phase 2 reentry and depolarization. Thus, the incidence and incidence of reperfusion arrhythmia can be alleviated.
3. The effects of SpreC and Post C on hemodynamics, myocardial enzyme release, myocardial infarction area and reperfusion arrhythmia were similar. The effects of SpreC and Post C on action potential, ICa, L, INa and Ito were similar.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R614;R-332
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