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; ICa,Lexposed 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 0.04mmol/L and 0.08mmol/L with the decreased value of 46.6%, 54.3% correspondingly. Further study of Anisodamine at 0.08mmol/L showed a marked inhibitory modulation of L-type Ca²⁺current density in a time-dependent manner with decreased ratio of (34.8±7.9) % (n=6, P<0.01), from the value of -4.474 pA/pF to - 2.882 pA/pF and accelerated Vi1/2of current inactivation curve from -14.7mV to -28.4mV and delayed Vi1/2of current activation curve from -15.6mV to -9.51mV. 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.