目的 研究參麥注射液對人誘導多能干細胞衍生心肌細胞（HiPSC-CM）鈣信號的影響。方法 通過CCK-8法測定參麥注射液（培養(yǎng)基中終體積分數為0.25%、0.50%、1.00%、2.00%）培養(yǎng)24h對HiPSC-CM細胞存活率的影響；采用鈣離子探針Fluo-4研究參麥注射液（2%，處理3min）對HiPSC-CM自發(fā)鈣瞬變頻率的影響，并在不同電刺激頻率（0.2、1.0、2.0Hz）下研究參麥注射液對HiPSC-CM鈣瞬變振幅、上升及下降時間和半峰全寬（FWHM）的影響。結果 CCK-8結果顯示，參麥注射液對HiPSC-CM無毒性，且在體積分數2%時細胞存活率顯著高于對照組（P＜0.05）。與加藥前比較，對照組加藥后自發(fā)鈣瞬變頻率沒有明顯變化，2%的參麥注射液組自發(fā)鈣瞬變頻率顯著降低（P＜0.001）。與加藥前比較，在0.2、1.0Hz電刺激頻率下，參麥注射液顯著降低HiPSC-CM的鈣瞬變振幅、上升及下降時間和FWHM（P＜0.01）；在2.0Hz電刺激頻率下，參麥注射液顯著降低HiPSC-CM的上升、下降時間和FWHM（P＜0.01）。結論 參麥注射液能夠降低HiPSC-CM自發(fā)鈣瞬變的頻率，且在不同頻率的電刺激下能降低HiPSC-CM的鈣瞬變振幅，表明鈣信號調控可能是參麥注射液心肌保護作用的潛在機制之一。

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

國家自然科學基金項目(81973698、81703942);中華中醫(yī)藥學會青年人才托舉工程項目(2019-QNRC2-B08)