为了测量细胞内ROS的产生,选择对数生长期的16hbe细胞以每孔9x1

In order to measure the production of ROS in cells, 16hbe cells in logarithmic growth phase were selected to inoculate 9×10^3 cells per well in 96-well plates, and a certain concentration of PM2.5 and PM2.5 model particles were infected for 48h. The cells were washed twice with PBS buffer, and 10 μM dichlorofluorescein diacetate (DCFH-DA) (Sigma Aldrich) was added and then incubated at 37° C. in the dark for 30 min. Finally, after washing with serum-free MEM three times to completely remove the DCFH-DA that did not enter the cells, the DCF fluorescence signal was measured with a microplate reader with excitation wavelength and emission wavelength of 488nm and 525nm, respectively.

In order to measure the production of ROS in cells, 16hbe cells of the long-term number of children were selected to be inoculated in 96-well plates at 9x10,3 per well, and infected with a certain concentration of PM2.5 and PM2.5 model particles 48h. Wash the cells twice with pBS buffer and add 10 m dichloroluorolyphosphonate (DCFH-DA) (Sigma Aldrich) and incubate 30min in the dark at 37 degrees C. Finally, dcF fluorescence signals were measured with microplate readers with excitation wavelengths and emission wavelengths of 488nm and 525nm, respectively, after serum-free MEM washed three times to completely remove DCFH-DA that had not entered the cell.

In order to measure intracellular ROS production, 16HBE cells in logarithmic growth phase were inoculated into 96 well plate with 9x10 ^ 3 cells per well, and were exposed to certain concentrations of PM2.5 and PM2.5 model particles for 48h. The cells were washed twice with PBS buffer solution and added with 10 μ m dichlorofluorescein diacetate (DCFH-DA) (sigma Aldrich) and incubated at 37 ℃ for 30 min. Finally, after three washings of serum-free MEM to completely remove DCFH-DA that did not enter the cells, the fluorescence signal of DCF was measured with a microplate reader with excitation wavelength of 488 nm and emission wavelength of 525 nm.<br>