Background:The decreased slowly activating delayed rectifier K+current(IKs)is the molecular basis of arrhythmia caused by myocardial hypertrophy.The aim of this study was to investigate the mechanism of IKs down-regul...Background:The decreased slowly activating delayed rectifier K+current(IKs)is the molecular basis of arrhythmia caused by myocardial hypertrophy.The aim of this study was to investigate the mechanism of IKs down-regulation related to the channel number,as well as the regulation of channel number under pathological conditions.Methods:The HEK 293 cell co-transfected with KCNQ1/KCNE1 genes was cultured conventionally.After the cells incubated with angiotensin II(AngII)(24 h),AngII(72 h),bisindolylmaleimide I(Bis),brefeldin A and dynasore,the effect and relevant mechanism of long term incubation of AngII on the IKs tail current and KCNQ1 channel total protein were investigated by electrophysiology method and western blotting.In the experiment,the Bis,brefeldin A and dynasore could inhibit the protein kinase C(PKC)activity,the forward transport of KCNQ1 channel and the endocytosis of KCNQ1 channel,respectively.At last,the Rab GTPases 11(Rab11)dominant-negative mutant dsRed-Rab11/S25N was infected into the cells to investigate the effect and relevant mechanism of long term AngII incubation on the IKs tail current and KCNQ1 channel total protein.Results:Our results showed that the decreased IKs tail current and the KCNQ1 channel total protein caused by long term AngII incubation were attenuated by Bis treatment,which inhibited PKC activity.In addition,the inhibited IKs tail current and KCNQ1 channel total protein were also alleviated by brefeldin A and dynasore treatment.At last,the expression of Rab11 dominant-negative mutant dsRed-Rab11/S25N could weak the inhibition of IKs tail current and the KCNQ1 channel total protein caused by long term AngII incubation.Conclusion:The long term incubation of AngII inhibited the IKs tail current and KCNQ1 channel total protein was achieved by PKC activation and the disorder of the channel trafficking by Rab11.展开更多
基金supported by the National Natural Science Foundation of China(NSFC)Youth Project(No.82204397 and No.22203063).
文摘Background:The decreased slowly activating delayed rectifier K+current(IKs)is the molecular basis of arrhythmia caused by myocardial hypertrophy.The aim of this study was to investigate the mechanism of IKs down-regulation related to the channel number,as well as the regulation of channel number under pathological conditions.Methods:The HEK 293 cell co-transfected with KCNQ1/KCNE1 genes was cultured conventionally.After the cells incubated with angiotensin II(AngII)(24 h),AngII(72 h),bisindolylmaleimide I(Bis),brefeldin A and dynasore,the effect and relevant mechanism of long term incubation of AngII on the IKs tail current and KCNQ1 channel total protein were investigated by electrophysiology method and western blotting.In the experiment,the Bis,brefeldin A and dynasore could inhibit the protein kinase C(PKC)activity,the forward transport of KCNQ1 channel and the endocytosis of KCNQ1 channel,respectively.At last,the Rab GTPases 11(Rab11)dominant-negative mutant dsRed-Rab11/S25N was infected into the cells to investigate the effect and relevant mechanism of long term AngII incubation on the IKs tail current and KCNQ1 channel total protein.Results:Our results showed that the decreased IKs tail current and the KCNQ1 channel total protein caused by long term AngII incubation were attenuated by Bis treatment,which inhibited PKC activity.In addition,the inhibited IKs tail current and KCNQ1 channel total protein were also alleviated by brefeldin A and dynasore treatment.At last,the expression of Rab11 dominant-negative mutant dsRed-Rab11/S25N could weak the inhibition of IKs tail current and the KCNQ1 channel total protein caused by long term AngII incubation.Conclusion:The long term incubation of AngII inhibited the IKs tail current and KCNQ1 channel total protein was achieved by PKC activation and the disorder of the channel trafficking by Rab11.
