Titanium dioxide nanoparticles induce apoptosis through ROS-Ca2+ -p38/AKT/mTOR pathway in TM4 cells

Titanium dioxide nanoparticles (TiO₂ NPs) can cause Apoptosis in TM4 cells; however, the underlying mechanism has not been entirely elucidated. The purpose of this study was to investigate the effects of TiO₂ NPs on ROS, CA²⁺ level, p38/Akt/mTOR pathway, and Apoptosis in TM4 cells and to evaluate the role of CA²⁺ in p38/Akt/mTOR pathway and Apoptosis. After exposure to different concentrations (0, 50, 100, 150, and 200 μg/mL) of TiO₂ NPs for 24 h, cell viability, ROS, CA²⁺ level, CA²⁺ -ATPase activity, p38/Akt/mTOR pathway-related proteins, Apoptosis rate, and apoptosis-related proteins (Bax, Bcl-2, Caspase 3, Caspase 9, and p53) were detected. The ROS scavenger NAC was used to determine the effect of ROS on CA²⁺ level. The CA²⁺ chelator BAPTA-AM was used to evaluate the role of CA²⁺ in p38/Akt/mTOR pathway and Apoptosis. TiO₂ NPs significantly inhibited cell viability, increased ROS level, and elevated CA²⁺ level while suppressing CA²⁺ -ATPase activity. TiO₂ NPs regulated the p38/Akt/mTOR pathway via increasing p-p38 level and decreasing p-AKT and p-mTOR levels. TiO₂ NPs significantly enhanced the Apoptosis. NAC attenuated CA²⁺ overload and reduction in CA²⁺ -ATPase activity caused by TiO₂ NPs. BAPTA-AM alleviated TiO₂ NPs-induced abnormal expression of p38/Akt/mTOR pathway-related proteins. BAPTA-AM assuaged the Apoptosis caused by TiO₂ NPs. Altogether, this study revealed that TiO₂ NPs elevated intracellular CA²⁺ level through ROS accumulation. Subsequently, the heightened intracellular CA²⁺ level was observed to exert regulation over the p38/Akt/mTOR pathway, ultimately culminating in Apoptosis. These results provides a complementary understanding to the mechanism of TiO₂ NPs-induced Apoptosis in TM4 cells.

Ca2+; TM4 cells; apoptosis; p38/AKT/mTOR pathway; titanium dioxide nanoparticles.