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<inf>Ca,L</inf>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 0.04mmol/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 V<inf>i1/2</inf>of current inactivation curve from -14.7mV to -28.4mV and delayed V<inf>i1/2</inf>of 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.
Tian-Nan
Wang
W
Dept. of Biomedical Engineering, Key Laboratory of Biomedical Engineering of Ministry of Education, Zhejiang University, Zheda Road 38, 310027 Hang Zhou, China.
Hui-Juan
Yang
Y
Gu-Ling
Jie-Yue
Li
L
Xiao-Xiang
Zheng
Z