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目的观察抗钠-钙交换体α-2(807-820)抗体对大鼠心功能影响并分析其机制。方法以化学合成的α-2(807-820)肽段作为抗原免疫大鼠制备并纯化抗体,利用Langendorff离体灌流系统观察其对大鼠心功能影响;利用全细胞膜片钳技术观察抗体对心肌细胞钠-钙交换电流作用。结果大鼠经主动免疫后,抗α-2(807-820)抗血清滴度明显升高,纯化后抗体浓度为6.3 mg/mL,聚丙烯酰胺凝胶电泳时仅有1条带出现;10和40 nmol/L的抗体可明显升高离体灌流大鼠心脏左室发展压、左室压最大上升速率和下降速率,这一效应可被KB-R7943阻断;膜片钳实验表明,抗α-2(807-820)抗体(40 nmol/L)可使外向和内向的钠-钙交换电流分别从给药前的(1.05±0.17)、(0.86±0.21)pA/pF增加至(1.53±0.28)、(1.23±0.31)pA/pF(P<0.05)。结论抗α-2(807-820)抗体通过激动心肌钠-钙交换体不仅可以增强心肌的收缩力和加快心肌收缩的速度,还可促进心肌舒张。
Objective To observe the effect of anti-sodium-calcium exchanger α-2 (807-820) on cardiac function in rats and analyze its mechanism. Methods Antibody was prepared from the chemically synthesized α-2 (807-820) peptide as an antigen and purified by Langendorff in vitro perfusion system. The effect of antibody on cardiac function was observed by whole cell patch clamp technique. Cell sodium-calcium exchange current effect. Results The titer of anti-α-2 (807-820) antiserum was significantly increased after active immunization. The antibody concentration after purification was 6.3 mg / mL. Only one band appeared when polyacrylamide gel electrophoresis was performed. 10 And 40 nmol / L antibody could significantly increase left ventricular pressure, maximum rate and rate of left ventricular pressure in isolated perfused rat hearts, and this effect could be blocked by KB-R7943. Patch-clamp experiments showed that the anti- The exogenous and inward sodium-calcium exchange currents increased from (1.05 ± 0.17), (0.86 ± 0.21) pA / pF before administration to (1.53 ± 0.31) in the presence of α-2 (807-820) ± 0.28), (1.23 ± 0.31) pA / pF (P <0.05). Conclusion The anti-α-2 (807-820) antibody can not only enhance myocardial contractility and myocardial contractile speed, but also promote myocardial relaxation by activating cardiac sodium-calcium exchanger.