Measuring method of cigarette smoke induced apoptosis under exposure of gas-liquid interface
A method of determination, technology of cigarette smoke, applied in the field of determination of apoptosis induced by cigarette smoke under the exposure of gas-liquid interface, to achieve the effect of real and intuitive results, simple operation, simple and accurate operation
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Embodiment 1
[0036] This embodiment provides a method for measuring the apoptosis induced by cigarette smoke under the exposure of the gas-liquid interface, specifically measuring the apoptosis of heat-not-burn cigarette smoke; the details are as follows:
[0037] (1) Configure experimental reagents: growth medium (DEME-H basal medium plus 10% FBS), exposure medium (DEME-H basal medium plus 10% FBS, 0.01M phosphate buffer solution (0.2g KCl, 0.2g KH2PO4, 8g NaCl, 2gNa 2HPO 4 12H 2 (0, add ultrapure water to 1L, measure pH to be 7.2, autoclave);
[0038] (2) Cell inoculation and culture: select mouse mononuclear macrophage RAW264.7 in the logarithmic growth phase, and use 1*10 6 The density of cells / well was inoculated on a 24 mm insert culture plate (Transwell culture plate), wherein 2 mL of growth medium was added to the lower chamber, and 1.5 mL of growth medium was added to the upper chamber, and cultured for 24 hours.
[0039] (3) Cigarette smoke exposure: After 24 hours of cell cu...
Embodiment 2
[0046] In this example, the cell apoptosis of 1 mg of cigarette smoke is measured, and the operation steps are the same as in Example 1; as the exposure dose increases, the cell apoptosis rate increases accordingly.
[0047] The apoptosis rate of 1mg cigarette smoke is shown in Table 2;
[0048] Table 2 Apoptosis rate of 1mg cigarette smoke (mean ± standard deviation)
[0049] smoke exposure dose 10％ 20％ 30％ 40％ 60％ Apoptosis rate (%) 1.9±0.4 11.7±2.2 23.6±1.4 34.2±1.4 76.5+1.1
Embodiment 3
[0051] In this example, the apoptosis of 3R4F cigarette smoke is measured, and the operation steps are the same as in Example 1; as the exposure dose increases, the apoptosis rate increases, showing a significant dose-effect relationship.
[0052] The apoptosis rate of 3R4F cigarette smoke is shown in Table 3;
[0053] Table 3 Apoptosis rate of 3R4F cigarette smoke (mean ± standard deviation)
[0054] smoke exposure dose 10％ 20％ 30％ 40％ 60％ Apoptosis rate (%) 2.4±2.8 14.2±3.2 73.1±3.1 86.3±2.9 97.9+1.4
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