Influence Of Titanium Ions On The Concentration Of Calcium Iron In T Cells

Objective:To study the effect of titanium ions on the calcium ion distribution of content and changes、the expression of inositol-1,4, 5-trisphosphate (IP3) and phospholipaseCyl (PLCyl) include its phosphorylation levels in T cells, and investigate the influence of titanium ions on the concentration of calcium ion in T cells and the calcium channel related protein of IP3 and PLCγ1.Methods:1. Jukat T cells were cultured with titanium ions. MTT assay was performed to detect the concentration of titanium when the activity of T cells were increased by 50%,95% and then select the maximum proliferation rate and the the middle of ion concentration, as the concentration of the experimental group.2. The Jukat T were divided into phytohemagglutinin(PHA) pre-activated group and non PHA pre-activated group, then both co-culture with different con-centrations of titanium ions. With laser scanning confocal microscope to detect the calcium ion distribution of content and changes after 24 hours, and use enzy -me-linked immunosorbent assay (ELLSA) to detect the expression of IP3 after 24 hours,and using Western blot (WB) to detect the protein levels and phosphor-ylation levels of PLCγ1 in T cells after 24 hours.Results:1. The concentration of titanium ions were lower than 100μM/L that make T cell activation and proliferation.With the increase of concentration, the proliferation rate rise. The concentration between 100-200μM/L, proliferation rate decrease while the concentration more than 200μM/L, cells were inhibition.The proliferation rate of 50%,95% were 5μM/L、10μM/L,and the maximum proliferation rate was 100μM/L, the intermediate concentration was 50μM/L.2. After treated with different concentration of titanium ions (5μM/L, 10μM/L,50μM/L,100μM/L),in the extracellular solution containing Ca2+, it can make intracellular calcium increased in the non pre-activated and pre-activated T cells, and more increase in pre-activated T cells. Compared with the control group, the intracellular calcium concentration significantly increased, the difference was statistically significant (p<0.05). In the extracellular solution without Ca2+, the intracellular Ca2+ concentration was not significantly changed in non pre-activated or pre-activated T cells,compared with the control group, the difference was not statistically significant (p>0.05).3. After treated with different concentration of titanium ions (5μM/L, 10μM/L,50μM/L, 100μM/L) the expression of IP3in the pre-activated T cells were siginificantly increased after 24h,compared with control group,the difference was statistically significant (p<0.05).Different concentrations show significantly difference(p<0.05).After treated with titanium ions, the expression of IP3 was decreased that without PHA pre-activated, compared with the control group, the difference was statistically significant (p<0.05). There were significantly difference compared with different concentrations (p<0.05).4. After treated with different concentration of titanium ions (5μM/L,10μM/L,50μM/L,100μM/L) the phosphorylation level of PLCyl were increased after 24h in the PHA pre-activated T cells, compared with the control group,the difference was statistically significant (p<0.05). There were significantly difference compared with different concentrations (p<0.05). After treated with titanium ions, the phosphorylation level of PLCyl was decreased that without PHA pre-activated, compared with the control group, the difference was statistically significant (p<0.05).Conclusions:1. titanium ions in concentration below 100μM/L can activate and proliferation T cell.2. Titanium ions can stimulate pre-actived T cells to increase the expression of Ca2+、IP3 and PLCyl phosphorylation level.