参麦注射液对人诱导多能干细胞衍生心肌细胞钙信号的影响

Effects of Shenmai Injection on calcium signaling in HiPSC-CM

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24 h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3 min)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、2.0 Hz)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、1.0 Hz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2.0 Hz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% and2.00%) 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.