Functional drug-loaded system used for treating lung tumour and preparation method and application thereof
A lung tumor and drug-carrying technology, applied in the field of lung tumor drug research, can solve problems such as tumor recurrence, achieve the effects of enhancing the effect of promoting apoptosis, realizing drug targeting, and reducing toxic and side effects
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Embodiment 1
[0066] Embodiment 1, the preparation of liposome
[0067] 1. Preparation of blank liposomes
[0068] 1. Preparation of blank liposomes
[0069] Dissolve lecithin, cholesterol and DSPE-PEG2000 in methanol at a molar ratio of 63.86:9.83:1, remove methanol by rotary evaporation, and form a film in an eggplant-shaped flask by rotary evaporation at 35°C and 40rpm, add 10mL of 250mM ammonium sulfate, and first place in a water bath Ultrasonic for 5 min, then transferred to a serum bottle and further ultrasonicated in a JY92 ultrasonic cell pulverizer (ultrasonic working time is 10 s, intermittent time is 30 s, the whole time is 10 min, and the protection temperature is 35 ° C), and then use liposome extrusion The press was passed through 400nm and 200nm polycarbonate membranes respectively 3 times, and the liposome external phase was replaced with PBS solution (pH=7.4) by dialysis to obtain blank liposomes.
[0070] 2. Preparation of blank liposomes modified by distearoylphosphati...
Embodiment 2
[0076] Embodiment 2, the performance measurement of liposome
[0077] 1. Determination of Encapsulation Efficiency
[0078] The unencapsulated ragragulin and EGGC were removed by dialysis, and the content was determined by high performance liquid chromatography. Take an appropriate amount of ragragulin liposomes, EGCG liposomes, FA-HPDL-EGCG liposomes and digest with acetonitrile before dialysis, and determine the contents of ragragulin and EGCG; , EGCG liposome, FA-HPDL-EGCG liposome plus acetonitrile digestion, measure the content of the EGCG entrapped in the liposome, calculate the drug encapsulation efficiency according to formula (1):
[0079] Encapsulation rate%=W 包 / W 总 ×100% (1)
[0080] where W 总 is the total drug dose; W 包 Indicates the amount of drug entrapped in liposomes.
[0081] In the prepared drug-loaded liposome, the encapsulation efficiency of ragragulin can reach 86%, and the encapsulation efficiency of EGCG can reach 75%.
[0082] 2. Particle size ...
Embodiment 3
[0091] Example 3. Research on the inhibitory effect of drug-loaded liposomes on lung adenocarcinoma A549 cells in vitro
[0092] 1. Training conditions
[0093] 1. Culture conditions of lung adenocarcinoma A549 cells
[0094] Lung adenocarcinoma A549 cells were cultured in DMEMF12 high-glucose medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin at 37 °C, 5% CO 2 Incubated in a saturated humidity incubator, the lung adenocarcinoma A549 cells grew adherently, and the cell morphology was spindle-shaped, ( figure 2 As shown in (A), the cells in the logarithmic growth phase were taken for the experiment.
[0095] 2. Culture conditions of lung adenocarcinoma A549 stem cells
[0096] A549 cells in the logarithmic growth phase were taken, washed with PBS, digested with 0.25% trypsin, blown into single cell suspension, resuspended in serum-free medium (containing DMEM / F12, bFGF, EGF, B27), 4~ After 5 days, the cells proliferate to form tumor st...
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