Effect Of Magnetic Nanoparticles On Kv1.3 Potassium Channel Of Jurkat Cells

Background:Nanobiology as a newly emerging multidisciplinary fused science has introduced a new dimension to research endeavor and a range of technologies in biology.As nanopaticles have particular physical property and the size of them are almost equal to protein,DNA and virus,the marvellous results may be present after nanoparticles coculture with cells.However,the mechanisms underlying nanoparticles influence the immune system remain obscure.The purpose of this research is to estabilish a new method to record Kv 1.3 current in Jurkat E6-1 cell accurately and study the potential molecular mechanism of SPION(superparamagnetic iron oxide nanoparticle)reacting with immunocyte.Materials and methods:Patch clamp was used to record Kvl.3 activation,inactivation and recovery currents in Jurkat E6-1cell and the results of SPION impacting Kv1.3 currtent were also observed by it.The Western blot and confocal system were chosen to detect the change of Kvl.3 protein expression under influence of SPION.Free[Ca2+]i free was detected lively by live cell imaging system.The apoptosis induced by SPION,which presented concentration-dependence,was measured by flow cytometry.Results:1.The electrophysiological characteristics and kinetics study of Kv1.3 in Jurkat cell:(1).It was necessary to dispose cells with 0.05%typsin before experiment,which was helpful to form giga seal.(2).Both the size of electrode caliber and[Ca2+]i free of internal solution were two key points in the process of recording current.The larger pore of electrode might lead to cell injured,or the whole-cell mode could not be made easily.The Ca2+ mixed in internal solution could disturb the recording results and the complex current might be got finally,and then EGTA was needed to chelate Ca2+ to avoid it.(4).Kv 1.3 showed its rapid activation(?act=6.47 ± 2.44ms)and inactivation(Tin=622.49±93.90ms),slow recovery after inactivation(Trec=6.141s)characteristics according to the results.2.The influence of Kv1.3,intracellular free calcium concentration and apoptosis under the SPION.(1).Kvl.3 could be inhibited by SPION in 15 minutes,which showed time-dependence.(2).The expression of Kv1.3 protein could also be suppressed by SPION,which presented concentration-dependence.(3).SPION could not be phagocytosed after incubation with Jurkat cell.(4).The elevation of[Ca2+]i and concentration-dependence apoptosis were observed due to effect of SPION.