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Sodium TanshinoneⅡ-A Sulfonate(DS-201) Modulated the Kinetics of BKCa withβ1 Subunit Independence in HEK-293 Cells

Sodium TanshinoneⅡ-A Sulfonate(DS-201) Modulated the Kineti...

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中国生理学会心血管生理学术研讨会

作者： Isao Inoue Key laboratory of medical electrophysiology of Ministry of Education Luzhou Medical College

Objective:We have investigated the unique effect of sodium tanshinoneⅡ-A sulfonate(DS-201) on the calcium dependency and kinetics of large conductance Ca2+-sensitive K+ channels (BKCa) on mouse cerebral arterial smooth muscle *** data suggest that DS-201 increases the calcium dependency and inhibits the transition of BKCa from the open to the closed *** further focusing on the kinetics of DS-201 on BKCa,in this study,we addressed the effect of DS-201 on BKCa heterogenetic expressed in HEK-293 cells by using patch-clamp ***:HEK293 cell strain with stable expression of BKCa was constructed through transfecting the plasmid pcDNA3.1-BKCa(a) and/or pEFl- BKCa(β1) by using lipofectamine *** cell strain was identified with patch clamp,western blot and *** currents were recorded in whole-cell and single channel configuration. The macro-currents of BKCa in inside-out configuration were recorded for analyzing the voltage dependency and kinetics of ***:(1) HEK293 cell strain with stable expression of BKCa was constructed *** basic electrophysiological characteristics of BKCa were consistent with those reported previously,*** conductance,voltage dependent- and calcium dependent- characteristics.200 nM IbTX also inhibited the currents in the whole-cell and outside-out *** specific band at 20 kD was detected with western blot and the specific expression was also detected with ***β1 subunit of BKCa increased the mean open time.(2) In whole-cell and single channel configuration,80μM DS-201 increased the macroscopic current density withβ1 subunit independence.(3) 80 and 160μM DS-201 increased the voltage dependence of BKCa by decreasing the half activated voltage(V1/2) from 102±20 mV(n=5) to 54±8 mV(P<0.05,n=5) and to 30.64 mV,*** also delayed the deactivation time constant from 0.27 ms to 0.91 ms(80μM DS-201) and to 10.72 ms(160μM DS-201) by analyzing the tail

关键词： BKCa DS-201 kinetical mechanism

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