Effect of Danhong Injection on electrophysiology of human-induced pluripotent stem cell-derived cardiomyocytes using patch clamp technique
Objective To explore the possible mechanism of Danhong Injection(DHI)in the treatment of aconitine-induced arrhythmia from the perspective of cellular electrophysiology through whole-cell patch-clamp experiments.Method Based on the patch-clamp technique of human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs),the inhibition effects of aconitine at different concentrations(3,9,27 μmol·L-1)on sodium channel and hERG potassium channel were observed,and the modeling conditions were screened.The control group,model group(1 μmol·L-1 aconitine for a long time)and DHI(3,9,27 μL·mL-1)groups were set up,and aconitine and DHI were added at the same time.The action potential amplitude(APA),discharge frequency,time-domain of 50％ action potential decrease(APD50)and time-domain of 90％ action potential decrease(APD90)were recorded,and the relative values were calculated using the control group as the standard.The control group,model group and DHI(3 μL·mL-1)group were set up,and aconitine and DHI were added at the same time,observed for 3-5 min,and the current densities of sodium ion,hERG potassium ion,and calcium ion were recorded.Results The inhibition rates of aconitine at 3,9,and 27 μmol·L-1 on sodium current were 17.94％,31.07％,and 60.67％,respectively,and the inhibition rates of hERG channel were 5.02％,10.59％,and 28.59％,respectively.The inhibition effect of aconitine on hERG channel in a short period of time was limited,while in the actual experiment,the long-term incubation of high concentration of aconitine completely inhibited the sodium current,resulting in the damage of ion channel function,so the model was prepared by 1 μmol·L-1 aconitine for a long time.Compared with the control group,the relative APA,relative APD50,and relative APD90 of hiPSC-CMs in the model group were significantly decreased(P＜0.01,0.001),and the relative discharge frequency was significantly increased(P＜0.001).The sodium current decreased within a certain time,with an inhibition rate of about 30.18％.The hERG potassium current decreased significantly,with an inhibition rate of about 72.33％.The calcium current increased significantly,with an increase of about 191.35％.Compared with model group,the relative discharge frequency of DHI 3,9,and 27 μL·mL-1 groups was significantly decreased(P＜0.05,0.001),and the relative APD90 was significantly increased(P＜0.05,0.01),and the relative APD50 of 3 μL·mL-1 group was significantly increased(P＜0.001).The decrease of sodium current,the decrease of hERG potassium current,and the increase of calcium current in the DHI group were alleviated to some extent.Conclusion DHI may alleviate aconitine-induced arrhythmias by influencing sodium,potassium,and calcium channels.
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