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目的:研究缺血再灌注(ischemia/reperfusion,I/R)期间不同时相(缺血时、缺血后再灌时)氟比洛芬酯处理对乳鼠心肌细胞钠电流的影响。方法:实验分为四组:I组(对照组)为体外培养乳鼠心室肌细胞;II组(I/R组)为相同的细胞,缺血3小时,再灌注1小时;III组(氟比洛芬酯缺血时处理组)在I/R缺血时相加入氟比洛芬酯(0.15μM);IV组(氟比洛芬酯缺血后再灌注时处理组)在I/R再灌注时相加入氟比洛芬酯(0.15μM)。采用膜片钳全细胞模式分别记录并比较四组的钠电流。结果:和I组相比,II组在每一指令电压上都增大钠电流;在测试电压为-20mV的条件下,钠电流峰值电流密度从-375.47±70.31(pA/pF)增至-557.11±127.86(pA/pF)(n=5,P<0.05);稳态激活半激活电压(V1/2)分别为-42.81±1.21mV和-31.49±2.40mV(n=5,P<0.05);稳态失活半激活电压(V1/2)分别为-74.24±5.89mV和-68.70±6.26mV(n=5,P<0.05);通道失活后恢复τ分别为25.69±19.47ms和7.53±3.24ms(n=5,P<0.05)。III、IV组钠电流峰值电流密度从-375.47±70.31(pA/pF)分别降至-208.80±121.44和-278.91±188.26(pA/pF)(n=5,P<0.05);通道稳态激活V1/2分别为-40.89±9.81mV和-39.49±3.70mV(n=5,P>0.05);稳态失活V1/2分别为-74.45±5.02mV和-75.49±3.32mV(n=5,P>0.05);通道失活后恢复τ分别为24.98±16.41ms和26.30±11.82ms(n=5,P>0.05)。III、IV组之间各指标间的差异无统计学意义。结论:I/R处理可以增大钠电流的峰值电流,并使电压电流曲线下移;减慢通道的时间依赖性激活,促进通道的电压依赖性失活,失活后恢复变快。而在I/R缺血和再灌注两个时相使用氟比洛芬酯均能有效阻制钠通道的这些改变。
OBJECTIVE: To study the effect of flurbiprofen axetil treatment on sodium current in neonatal rat cardiomyocytes during ischemia / reperfusion (I / R) phase at different phases (ischemia, reperfusion). Methods: The experiment was divided into four groups: group I (control group) were cultured neonatal rat ventricular myocytes; group II (I / R group) were the same cells, ischemia for 3 hours and reperfusion for 1 hour; group III Flurbiprofen axetil (0.15 μM) at I / R ischemia and group IV (flurbiprofen axetil reperfusion at reperfusion) in I / R Flurbiprofen axetil (0.15 μM) was added during reperfusion. Whole-cell patch clamp mode was used to record and compare the sodium currents in the four groups. Results: Compared with group I, group II increased the sodium current for each command voltage. When the test voltage was -20mV, the peak current density of sodium current increased from -375.47 ± 70.31 (pA / pF) to - 557.11 ± 127.86 (pA / pF) (n = 5, P <0.05). The steady state activation half activation voltage (V1 / 2) were -42.81 ± 1.21mV and -31.49 ± 2.40mV ). The half-activation voltage (V1 / 2) of steady-state inactivation were -74.24 ± 5.89mV and -68.70 ± 6.26mV respectively (n = 5, 7.53 ± 3.24ms (n = 5, P <0.05). The peak current densities of sodium currents in groups III and IV decreased from -375.47 ± 70.31 (pA / pF) to -208.80 ± 121.44 and -278.91 ± 188.26 (pA / pF) (n = 5, P <0.05) V1 / 2 were -40.89 ± 9.81 mV and -39.49 ± 3.70 mV, respectively (n = 5, P> 0.05); steady-state inactivation V1 / 2 was -74.45 ± 5.02 mV and -75.49 ± 3.32 mV , P> 0.05). The recovery τ of the channel after inactivation were 24.98 ± 16.41 ms and 26.30 ± 11.82 ms respectively (n = 5, P> 0.05). There was no significant difference between the indexes of III and IV groups. Conclusion: I / R treatment can increase the peak current of sodium current and decrease the voltage-current curve. It can slow down the time-dependent activation of channels and promote the voltage-dependent inactivation of channels. However, the use of flurbiprofen axetil at both I / R ischemia and reperfusion stages effectively blocked these changes in sodium channels.