Limonium franchetii antineoplastic active constituent extract and preparation method and application thereof
An effective component and anti-tumor drug technology, applied in the field of Yantai Limonium Limonium extract and its preparation, can solve the problems such as the preparation method and use of Yantai Limonium Limonium extract, and achieve the purpose of improving medicinal value, expanding uses, and expanding The effect of raw material channels
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Embodiment 1
[0026] Example 1: Preparation of the extract of effective anti-tumor parts of Yantai Limonium Herb
[0027] Dry and crush the whole herb of Limonium yantai, take 1500g of the mixture, add 8-10 times the amount of 95% ethanol to soak for 2-3 days, extract the alcohol-soluble components, repeat three times, combine the extracts three times, and reduce pressure at 30°C~40°C Recover ethanol until there is no alcohol smell to obtain the total extract, add distilled water and stir to form a suspension evenly, put it into a separatory funnel, add an equal volume of cyclohexane to extract three times, combine the cyclohexane extracts, recover under reduced pressure, and store at 40°C Dry in a vacuum dryer to constant weight to obtain cyclohexane fractions. After removing the remaining cyclohexane under reduced pressure, the remaining aqueous solution was extracted three times with an equal volume of ethyl acetate, the ethyl acetate extracts were combined, recovered under reduced press...
Embodiment 2
[0028] Example 2: Research on Antitumor Activity of Yantai Limonium Limonium Extract
[0029] Taking the extract of the effective antitumor part of Yantai Limonium Limonium obtained in Example 1 as the test drug, various tumor cells in the logarithmic growth phase were taken (the tumor cell lines used have the following types: human leukemia cell line HL-60 and K562, human liver cancer cell line HepG2, human lung adenocarcinoma cell line A549, human breast cancer cell line MCF-7, human cervical cancer cell line Hela), adjust the cell concentration with RPMI-1640 complete culture medium containing 10% calf serum , 180 μL of 8000 cells per well were inoculated in a 96-well cell culture plate, placed at 37 °C, 5% CO 2 overnight in the incubator, and add medicine the next day. The sample solutions from different extraction sites were prepared into a mother solution with an initial concentration of 1 mg / mL, double-diluted, and 20 μL of the diluted drug was added to each well, and...
Embodiment 3
[0030] Example 3: Effect of Yantai Limonium Limonium Extract on Induced Differentiation of Acute Myeloid Leukemia HL-60 Cells
[0031] Taking the extract of the effective anti-tumor part of Yantai Limonium Limonium obtained in Example 1 as the test drug, the HL-60 cells in the logarithmic growth phase were treated with the extracts of different parts at a final concentration of 25 μg / mL, and 5 × 10 4 Cells / mL were plated in 24-well plates and stored at 37°C, 5% CO 2 Cultured in the incubator for 5 days, collected cells in each group, smeared, Wright stained, observed cell morphology under a microscope and took pictures to observe the characteristics of cell size, nuclear-to-cytoplasmic ratio, nuclear shape, and cytoplasmic staining. The results showed that the leukemic HL-60 cells were treated with extracts from four parts of Limonium yantai for 5 days, and the cell morphology changed significantly under the light microscope (see figure 1). It shows that the nuclear chroma...
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