Application of Low Temperature Plasma Combined with Metformin
A technology of low-temperature plasma and metformin, which is applied to parts of surgical instruments, medical preparations containing active ingredients, drug combinations, etc., can solve problems such as unclear effects and achieve the effect of inducing cell death
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Embodiment 1
[0021] Example 1: Determination of the inhibitory effect of low temperature plasma (CAP) and metformin (MET) alone or in combination on U251 / U87 glioma cells
[0022] 1. In order to determine whether the independent action of low-temperature plasma and metformin has an effect on the cell viability of the glioma cell line, in this embodiment, the effects of low-temperature plasma and metformin on the viability of the glioma cell line were first detected respectively. The U251 cell line was divided into 7.5*10 4 cells / ml, U87 cell line 5*10 4 Inoculated in 96-well plate at a density of / ml, after 24 hours of culture:
[0023] (1) Plasma alone effect experiment: according to the low-temperature plasma treatment time of 10, 20, 40, and 60 seconds, respectively, after 24 hours and 48 hours of treatment, the cell viability was detected by the CellTiter Blue method. The results are as follows figure 1 As shown in A-1B, it can be seen from the results in the figure that low-temperat...
Embodiment 2
[0029] Example 2: Determination of the effect of the combination of low temperature plasma and metformin inducing the death of U251 / U87 glioma cells
[0030] After culturing U251 and U87 glioma cells in a 96-well plate for 12 hours, add 16 mmol / L of metformin, continue culturing for 12 hours, and then treat them with low-temperature plasma for 30 seconds, and continue culturing for 12 hours. The results are as follows: figure 2 As shown in A-2B; it can be seen from the figure that the combination of low-temperature plasma and metformin can change the shape of the cells, making the cells round and showing a death-like shape. As detected by the cell viability kit, compared with a single treatment, the combined use can indeed significantly induce cell death (such as figure 2 C-2D, living cells are in the circle, and dead cells are in the square; figure 2 E-2F is the statistical graph of cell death rate).
[0031] figure 2 In A-2B, the glioma cell line U251 ( figure 2 A),...
Embodiment 3
[0034] Example 3 Carry out further analysis experiments on the synergistic inhibitory effect mechanism of low temperature plasma and metformin
[0035] U251 and U87 glioma cells were cultured in a 96-well plate for 24 hours, then treated with low-temperature plasma for 30 seconds, and the level of free oxygen in cells was detected with a reactive oxygen species detection kit and detected with a hydrogen peroxide kit. Hydrogen peroxide levels in low-temperature plasma-treated cell culture media.
[0036] Such as image 3 A, glioma cell lines U251 (above) and U87 cell lines (below) were cultured for 12 hours, treated with CAP for 30 seconds, and the level of free oxygen in cells was detected with a reactive oxygen species detection kit (inside the circle is intracellular active oxygen), and found that CAP treatment significantly increased the level of free oxygen in cells.
[0037] Such as image 3 B. The hydrogen peroxide level in the CAP-treated cell culture medium was dete...
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