The application of the active extract of the fruit of the wood turtle in the preparation of anti-respiratory system tumor drugs
A kind of Momordicae semen and extract technology, which is applied in the field of pharmacy and can solve problems such as no reports on Momordicae semen extract.
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Embodiment 1
[0020] Embodiment 1 prepares the active extract of semen japonicus
[0021] Grind the Chinese medicinal materials of Saccharia chinensis to obtain the middle-grade powder of Sorcus chinensis, take 100g of the middle-aged powder of Sorcus chinensis, extract with 500ml of sodium acetate solution (20mM, pH 5.0) in the refrigerator at 4°C for overnight, centrifuge and freeze-dry the filtrate to obtain the extract , stored in -20°C refrigerator for later use. After dissolving the freeze-dried powder of the extract of the fruit of the tree turtle obtained by extraction in water, centrifugal filtration removes a small amount of insoluble impurities, and further separation is carried out by chromatography (such as figure 1 Shown), the chromatographic conditions are: chromatographic column: YMC-Pack ODS-A (150 * 10mm, 5 μ m); Column temperature: room temperature; Mobile phase: A: 0.05% formic acid B: 90% acetonitrile+10% water (containing 0.045 % formic acid); flow rate: 2ml / min; inje...
Embodiment 2
[0025] Example 2 In vitro anti-tumor effect test of the fruit of the fruit of the tree
[0026] The A549 cells in the logarithmic growth phase were inoculated into 96-well plates, and divided into blank control group, carboplatin control group and wood turtle extract treatment group (0.01, 0.05, 0.1, 0.2, 0.4mg / ml), 37 ℃ After culturing for 48 hours, MTT assay was performed to calculate the cell proliferation rate. The results showed that with the increase of dose, the extract of Sapia chinensis significantly inhibited the proliferation of A549 cells (such as figure 2 shown).
[0027] Example 2. Experiment of the effect of the extract of the fruit of the fruit of the tree on the inhibition of the growth cycle of A549 cells
[0028] The A549 cells in the logarithmic growth phase were inoculated into 96-well plates, and after being treated for 48 hours with the extract of Saccharina chinensis, the cells were collected by centrifugation, washed twice with PBS, and fixed overni...
Embodiment 3
[0029] Example 3. Experiment of the effect of the extract of the fruit of the fruit on the induction of A549 cell growth and apoptosis
[0030] A549 cells were inoculated in 24-well plates, and after being treated for 48 hours with the extract of the fruit of the wood beetle, the Hoechst33342 solution with a final concentration of 5 μg / ml was added, and placed at room temperature in the dark for 5 minutes. Discard the culture medium, fix with 4% formaldehyde for 10 min, discard by suction, wash with PBS 1-2 times, and add 1ml of PBS. The size and shape of apoptotic cells were observed under a fluorescence microscope. In addition, A549 cells were collected by centrifugation, rinsed once with pre-cooled PBS, added AnnexinV-FITC, mixed evenly, and incubated at room temperature in the dark for 10 min. Wash once with PBS, add appropriate amount of PI, and perform quantitative detection of cell apoptosis by flow cytometry. The results showed that the extract of the fruit of the fr...
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