Extractive of combined prescription of ophiopogon deccotion and phragmitis decoction and application thereof in preparing medicament for restraining proliferation of lung adenocarcinoma cell A549
A technology of Ophiopogon japonicus decoction and extract, which can be used in drug combinations, medical preparations containing active ingredients, antitumor drugs, etc., can solve the problems of poor curative effect, high drug resistance, and unsatisfactory effects of radiotherapy.
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Embodiment 1
[0047] Example 1: Materials and methods.
[0048] 1. Main reagents
[0049] Rotary evaporator (Nanjing Jinzheng Teaching Instrument Co., Ltd., 1002 lifting constant temperature water bath), water circulation vacuum pump (Yingyu Yuhua Instrument Factory, Gongyi City, Henan Province), digital display thermostat heating mantle (Zhangzhishan Town, Tongzhou City), HH-S Digital display constant temperature water bath (Jintan Medical Instrument Factory), round bottom flask (20L), condenser tube, iron stand, separatory funnel (5000ml) (Nanjing Jinzheng Teaching Instrument Co., Ltd.).
[0050] Ophiopogon japonicus (Sichuan), French pinellia (Guizhou), sun-dried ginseng (Jilin), licorice (Inner Mongolia), reed stem (Anhui), coix seed (Guizhou), winter melon seeds (Anhui), peach kernel (Shandong).
[0051] Medicinal ethanol (analytical grade, Nanjing Chemical Reagent Co., Ltd.), cyclohexane (analytical grade, Shanghai Shisi Hewei Chemical Co., Ltd.), ethyl acetate (analytical grade, Nan...
Embodiment 2
[0055] Example 2: Preparation of the extract of Maimendong decoction combined with Qianjinweijing decoction.
[0056]Weigh 200g of Ophiopogon japonicus, 100g of Pinellia chinensis, 200g of raw sun-dried ginseng, 80g of licorice, 200g of reed stem, 200g of coix seed, 200g of winter melon seed, 100g of peach kernel, and make up a total of 1280g of the whole formula, crush it and put it in a 20,000mL round bottom flask , add 10 times the amount of medicinal materials in pure water, shake and mix, soak for 30 minutes; connect the reflux condensing device, put it in the electric heating mantle and heat slowly to 110°C to boil, and keep boiling slightly for 2 hours, stop heating, and place it to the solution temperature Cool down to room temperature, remove the reflux condenser, pour out the solution in the flask, and filter it with gauze. Add purified water to the dregs to the same scale as the first time, extract once more in the same way, combine the extracts, and concentrate und...
Embodiment 3
[0057] Example 3: Experiment of the inhibitory effect of the ethyl acetate extraction fraction (MW-06) on A549 cells.
[0058] 1. Experimental method
[0059] 1.1 Cell proliferation inhibition test (MTT) method
[0060] Digest and collect A549 cells and HFL-1 cells in the logarithmic growth phase, and use 5×10 3 Inoculate each well in a 96-well plate. After the cells are fused to 80%, replace the serum-free medium and culture for 12 hours to synchronize the cells. After that, add the final concentration of 500 μg / mL and 100 μg / mL diluted with RPMI1640 medium to the experimental group. , 10 μg / mL and 1 μg / mL of MW-06 sites, 200uL per well, set equal volumes of DMSO-containing control group (i.e. final concentration 5‰) and DMSO-free blank control group, each set 8 parallel wells , continue to culture for 72h, after observing under the microscope (×400), add 20μL of 5mg / mL MTT prepared in PBS at 37°C, 5% CO 2 Incubate in the incubator for 4 hours, discard the supernatant and ...
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