Advanced Measurement and Quantitative Appraise of Anisodamine on Calcium Triggered in Cardiac Myocyte

Both patch clamp and laser scanning confocal microscopy (LSCM) was applied to appraise the Anisodamine´s cardioprotective effects quantitatively and its mechanism were studied. MTT measurement was observed cell viability and Fluo-3/AM was utilized for real-time free calcium with LSCM; I_Ca,L exposed to Anisodamine was measured by whole-cell patch clamp recording technique. Our study observed that KCL-triggered calcium elevation could be decreased by Anisodamine at dose of 0.04 mmol/L and 0.08 mmol/L with the decreased value of 46.6%, 54.3% correspondingly. Further study of Anisodamine at 0.08 mmol/L showed a marked inhibitory modulation of L-type Ca²⁺ current density in a time-dependent manner with decreased ratio of (34.8 plusmn 7.9) % (n = 6, P < 0.01), from the value of -4.474 pA/pF to -2.882 pA/pF and accelerated V_il/2 of current inactivation curve from -14.7 mV to -28.4 mV and delayed V_il/2 of current activation curve from -15.6 mV to -9.51 mV. Our result suggests that Anisodamine conferred the cardioprotection by decreasing calcium elevation quantitatively; the likely mechanism was suggested to be responsible for inhibition of L-calcium channel