Application of crude extract of radix tetrastigme in aspect of inhibiting pancreatic cancer
A technology of crude extract and clover leaf, applied in biochemical equipment and methods, medical preparations containing active ingredients, drug combinations, etc., can solve the problems of low response rate, adverse reactions of chemotherapy drugs, etc., and achieve good separation Effect
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Embodiment 1
[0029] A method for obtaining the crude extract of Clover, specifically comprising the following steps:
[0030] S1. Take 155 g of the medicinal material powder of A. clover root, add 70% ethanol to carry out reflux extraction for 3 times, the reflux temperature is 80 degrees Celsius, and extract for 1.5 hours each time;
[0031] S2. Filter the solution after each extraction in step S1 with a 5 μm filter membrane to separate the extract and filter residue, combine the extracts, concentrate and dry the filtrate at 50° C., and obtain a solid crude extract. The weight of the solid crude extract is 14.17 g ;
[0032] S3, column chromatography points
[0033] S3-1. Macroporous adsorption resin: Take 600ml of HP-20 resin and soak it overnight with 95% ethanol; after suspension, install it into a glass chromatography column; first elute with 1.8L of 95% ethanol, and then equilibrate with water;
[0034] S3-2, take 13.5g of the solid crude extract of step S2, dissolve it with about ...
Embodiment 2
[0053] The death of pancreatic ductal adenocarcinoma (PDAC) cells was induced by the crude extract of Cloverleaf obtained in Example 1, which specifically included the following steps: (1) four kinds of PDAC cells were planted in 96-well plates, 5 × 10 cells per well. 3 cells; (2) after 24 hours, the cells adhered to the wall, and four components of the crude extract of Clover were added respectively: component-1 (10% (1-2) EtOH extract), component-2 (10 %(-3)-30%(1-2) EtOH extract), fraction-3 (30%(-1)-50%(-1)EtOH extract), fraction-4 (50%(- 2) -95% (-2) EtOH extract), the concentration gradient is set to be 0, 100, 250mg / L; (3) After 48 hours, add 10ul of PI dye of 0.05mg / ml to each well, and the medium in each well is 200ul system, the cell survival rate after treatment at a specified concentration was measured with a cell counting kit-8 (CCK-8).
[0054] After enrichment and separation, it was divided into four fractions: fraction-1 (10% (1-2) EtOH extract), fraction-2 (1...
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