Method for extracting anti-tumor active component from traditional Chinese medicine pericarpium juglantis
An anti-tumor activity, Qinglong technology
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Embodiment 1
[0021] Preparation of active components of traditional Chinese medicine Qinglongyi:
[0022] Material source: Qinglongyi, a traditional Chinese medicine, is the immature dry exocarp of Juglans regia L., which was collected in the suburb of Tianshui City, Gansu Province in August.
[0023] Extraction and preparation: 1 kg of dried Qinglongyi, 5L of 70% ethanol was heated and refluxed at 80°C for 3 times, each time for 2 hours, filtered, combined for 3 extractions, concentrated (vacuum-0.07MPa, temperature 50°C), reduced Dry under pressure (vacuum-0.07MPa, temperature 50°C) to obtain extract I. After dispersing the extract I with 20 times the amount of water, extract 3 times with 2 times the amount of ethyl acetate of the extract I solution, combine the ethyl acetate extracts, concentrate, and dry under reduced pressure (vacuum -0.07MPa, temperature 50°C) , to obtain extract II. Extraction II is dissolved with 10 times the amount of 20% ethanol, the supernatant is taken, and t...
Embodiment 2
[0025] inhibit mouse S 180 and H 22 Assay for Growth Activity of Transplanted Tumors
[0026] Test animals: Kunming mice, male, weighing 18-22 g.
[0027] Drugs and reagents: physiological saline for injection (blank control), cyclophosphamide (positive control), active components of Qinglongyi. Tumor strain: mouse solid tumor S 180 and mouse hepatocarcinoma H 22 .
[0028] experiment method:
[0029] Effects on tumor weight and immune organ index in mice: Male mice were randomly divided into Qinglongyi active group, cyclophosphamide group and normal saline group. Tumor cell suspension (approximately 1×10 6 ~2×10 6 cell / mL) 0.2 mL was inoculated under the skin of the right anterior armpit of the mice, and 24 hours after the inoculation, each group was intragastrically administered (i.g.) for 10 consecutive days. After the last administration, the mice were killed, and the body weight, tumor weight, thymus and spleen mass were weighed, and the tumor inhibition rate, th...
Embodiment 3
[0039] Experiment of Inhibiting Proliferation Activity of Human Lung Cancer A549 Cell Line
[0040] Cell culture:
[0041]Human lung adenocarcinoma A549 cells were cultured in RPMI 1640 medium, plus 10% FBS. Take the cell count in the logarithmic growth phase, and take 2.5×10 4 / mL concentration in 96-well plate, 200 μL per well, in 5% CO 2 , 95%O 2 , 37°C, cultured in a saturated humidity cell incubator for 24 hours until the cells adhered to the wall.
[0042] Cytotoxicity Assay:
[0043] After the drug was dissolved in DMSO, it was diluted to different concentrations (3.125-100 μg / mL) with 1640 culture medium. 2 , 95%O 2 , culture in a saturated humidity incubator at 37°C for 48 hours, discard the cell solution, add 200 μL of 1640 culture solution and 20 μL of 5% MTT to each well and continue to incubate for 4 hours, discard the cell solution, add 150 μL DMSO to each well, and shake for 10 minutes to make the crystals fully After dissolving, use a microplate reader t...
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