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目的分离大鼠肺动脉平滑肌细胞(PASMCs)并建立记录电压门控性钾离子通道(KV)和钙离子激活性钾通道(KCa)电流的方法。方法急性酶解法分离出单个PASMC,采用全细胞膜片钳技术记录KV和KCa电流。结果成功获得单个PASMC。在+60mV时,4-氨基吡啶5mmol/L可明显抑制KV电流,电流密度从(134.69±4.73)pA/pF减少到(35.02±5.64)pA/pF(P<0.05)。在+60mV时,四乙铵1mmol/L可明显抑制KCa电流,电流密度从(15.10±1.62)pA/pF减少到(3.82±0.72)pA/pF(P<0.05)。结论急性酶解法成功获得活性良好的PASMCs,并建立了KV、KCa电流的记录方法,为研究肺动脉高压时钾通道的电生理学改变奠定基础。
Objective To isolate rat pulmonary artery smooth muscle cells (PASMCs) and establish a method of recording voltage-gated potassium channel (KV) and calcium-activated potassium channel (KCa) currents. Methods A single PASMC was isolated by acute enzymatic digestion, and KV and KCa currents were recorded using whole-cell patch clamp technique. Results A single PASMC was successfully obtained. At +60 mV, 5 mmol / L 4-aminopyridine significantly inhibited the KV current, and the current density decreased from (134.69 ± 4.73) pA / pF to (35.02 ± 5.64) pA / pF; At +60 mV, 1 mmol / L of tetraethylammonium significantly inhibited the KCa current, and the current density decreased from (15.10 ± 1.62) pA / pF to (3.82 ± 0.72) pA / pF (P <0.05). Conclusions Acute enzymatic hydrolysis of PASMCs with good activity was achieved, and KV and KCa currents were recorded, which laid the foundation for the study of electrophysiological changes of potassium channels in pulmonary hypertension.