The Kv1.3 Of Voltage-gated Potassium Channel Effect Of Titanium Ions On The Change Intracellular Ca²⁺ Concentration Of T-lymphocytes

Objective: After adding the ShK-Dap22,to investigate the effects of titanium ions on the changes of intracellular calcium concentration,proliferation and Kv l.3 channel in Jurkat T cells.Methods: 1.After Jurkat T cells were cultured with ShK-Dap22,MTT assay was performed to determine the optimum concentration of ShK-Dap22.2.The Jurkat T cells were divided into two groups:?1?T cells stimulated by Sh K-Dap22?2?resting T cells.After co-culture with different levels of titanium ions,the flow cytometry was used to detect the distribution of calcium cations,cell membrane and content changes.the real-time fluorescent quantitative RT-PCR was used to detect the expression of Kv l.3 mRNA and content changes.Results: 1.When the concentration of Sh K-Dap22 was lower than10nmol/L,T cells can be induced to activate and restrain.With the increase of concentration,the restrain rate rise.When the concentration is in excess of10nmol/L,restrain rate stabilization.The optimum concentration were10nmol/L when T cells restrain rate reaches.When the concentration is up to10nmol/L,the lymphocyte restrain of ShK-Dap22 treated group was the most significantly comparing to that of control.2.After treated with different concentration of titanium ions?25?mol/L,50?mol/L,100?mol/L?,in the extracellular solution without Sh K-Dap22,the expression of Kv l.3mRNA increased in resting cells group and activated cells group,and the rise within activated T cells group is much more evident.Compared with the control group,the expression of Kv l.3 mRNA significantly increased,the difference was statistically significant?p <0.05?.In absence of the extracellular solution containing ShK-Dap22,the expression of Kv l.3mRNA was not significantly changed in both experimental groups,compared with the control group,the difference was not statistically significant?p >0.05?.After treated with different concentration of titanium ions?25?mol/L,50?mol/L,100?mol/L?,in the extracellular solution without Sh K-Dap22,the intracellular calcium concentration increased in resting cells group and activated cells group,and the rise within activated T cells group is much more evident.Compared with the control group,the intracellular calcium concentration significantly increased,the difference was statistically significant?p <0.05?.in absence of the extracellular solution containing ShK-Dap22,the intracellular Ca²⁺ concentration was not significantly changed in both experimental groups,compared with the control group,the difference was not statistically significant?p >0.05?.Conclusions: 1.ShK-Dap22 can inhibits T-cell proliferation.2.There was a lot of mechanism to effect the change intracellular Ca²⁺concentration of T-cell on titanium ions stimulating,Kv l.3 mechanism is one of them.