目的 研究参麦注射液对人诱导多能干细胞衍生心肌细胞（HiPSC-CM）钙信号的影响。方法 通过CCK-8法测定参麦注射液（培养基中终体积分数为0.25%、0.50%、1.00%、2.00%）培养24h对HiPSC-CM细胞存活率的影响；采用钙离子探针Fluo-4研究参麦注射液（2%，处理3min）对HiPSC-CM自发钙瞬变频率的影响，并在不同电刺激频率（0.2、1.0、2.0Hz）下研究参麦注射液对HiPSC-CM钙瞬变振幅、上升及下降时间和半峰全宽（FWHM）的影响。结果 CCK-8结果显示，参麦注射液对HiPSC-CM无毒性，且在体积分数2%时细胞存活率显著高于对照组（P＜0.05）。与加药前比较，对照组加药后自发钙瞬变频率没有明显变化，2%的参麦注射液组自发钙瞬变频率显著降低（P＜0.001）。与加药前比较，在0.2、1.0Hz电刺激频率下，参麦注射液显著降低HiPSC-CM的钙瞬变振幅、上升及下降时间和FWHM（P＜0.01）；在2.0Hz电刺激频率下，参麦注射液显著降低HiPSC-CM的上升、下降时间和FWHM（P＜0.01）。结论 参麦注射液能够降低HiPSC-CM自发钙瞬变的频率，且在不同频率的电刺激下能降低HiPSC-CM的钙瞬变振幅，表明钙信号调控可能是参麦注射液心肌保护作用的潜在机制之一。

Objective To study the effect of Shenmai Injection (SMI) on calcium signaling of human induced pluripotent stem cellderived cardiomyocyte (HiPSC-CM).Methods The effect of SMI (final volume fractions in medium were 0.25%, 0.50%, 1.00% and 2.00%) on the cell viability of HiPSC-CM was determined by CCK-8 assay. Calcium ion probe Fluo-4 was used to study the effect of SMI (2%, 3 min) on spontaneous calcium transient frequency in HiPSC-CM. The effect of SMI on amplitude, rise and decay time and half-peak full width (FWHM) of electrical stimulation-induced calcium transient in HiPSC-CM was investigated at different electrical stimulation frequencies (0.2, 1.0 and 2.0 Hz).Results The results of CCK-8 showed that SMI was not toxic to HiPSC-CM, and cell viability was significantly higher than that of control group at 2% (P <0.05). Compared with before treatment, the spontaneous calcium transient frequency in control group did not change significantly after treatment, and the spontaneous calcium transient frequency in 2% SMI group was significantly decreased (P <0.001). Compared with before treatment, SMI significantly decreased the calcium transient amplitude, rise and fall time and FWHM of HiPSC-CM at 0.2 and 1.0 Hz electrical stimulation frequency (P <0.01). At the frequency of 2.0 Hz electrical stimulation, SMI significantly reduced the rise and fall time and FWHM of HiPSC-CM (P <0.01).Conclusions SMI can decrease the frequency of spontaneous calcium transients in HiPSC-CM. Under the electrical stimulation at different frequency, SMI can reduce the amplitude of calcium transient in HiPSC-CM. It was indicated that SMI achieve its cardioprotective effect by regulating the calcium signal of cardiomyocytes.

R285.5

国家自然科学基金项目(81973698、81703942);中华中医药学会青年人才托举工程项目(2019-QNRC2-B08)