Biological detection method for Chinese medicine composition
A detection method and composition technology, applied in the field of medicine, can solve problems such as the inability to reflect the overall drug efficacy and the like
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Embodiment 1
[0162] Prescription: 255g forsythia, 255g honeysuckle, 85g roasted ephedra, 85g fried bitter almond, 255g gypsum, 255g isatis root, 255g Mianma Guanzhong, 255g houttuynia cordata, 85g patchouli, 51g rhubarb, 85g rhodiola, mint Brain 7.5g, licorice 85g;
[0163]Preparation method: the above thirteen flavors, patchouli plus water distillation to extract the volatile oil, collect the volatile oil, filter the water extract, and set aside; forsythia, ephedra, houttuynia, rhubarb are extracted twice with 70% ethanol, the first time 2 1.5 hours for the second time, filter the extract, combine, recover ethanol, and set aside; add honeysuckle, gypsum, isatis root, Mianma Guanzhong, licorice, and rhodiola to boiling, add fried bitter almonds, and cook for two 1.5 hours for the first time, 1 hour for the second time, filter the decoction, combine the filtrates, add the aqueous solution after patchouli oil extraction, concentrate to a relative density of 1.10-1.15 (60°C), add ethanol to m...
Embodiment 2
[0169] Bioassay method:
[0170] Step a, establishment of inflammation model: RAW264.7 cells in the logarithmic growth phase were digested with 0.25% trypsin, and the cell concentration was adjusted to 1×10 4 cells / well, seeded in 96-well plate, placed in 5% CO 2 , Cultivate in a 37°C incubator for 24 hours, discard the supernatant, wash twice with PBS, add 200 μL normal medium to the blank well, add 200 μL LPS medium with a final concentration of 1.0 μg / mL and a final concentration of 200 ng / mL to the model well IFN-γ medium, the drug group was given a certain concentration of traditional Chinese medicine composition under the inflammation model, placed in 5% CO 2 , Cultivate in a 37°C incubator for 24 hours. Aspirate the cell supernatant for the determination of the expression level of IL-6 secreted by the cells;
[0171] Step b, measuring the concentration of the inflammatory factor IL-6 in the cells: measuring the content of IL-6 in the cell supernatant by an immunoradi...
Embodiment 3
[0174] Bioassay method:
[0175] Step a, establishment of inflammation model: RAW264.7 cells in the logarithmic growth phase were digested with 0.25% trypsin, and the cell concentration was adjusted to 1×10 4 cells / well, seeded in 96-well plate, placed in 5% CO 2 , Cultivate in a 37°C incubator for 24 hours, discard the supernatant, wash twice with PBS, add 200 μL normal medium to the blank well, add 200 μL LPS medium with a final concentration of 1.0 μg / mL and a final concentration of 200 ng / mL to the model well IFN-γ medium, the drug group was given a certain concentration of traditional Chinese medicine composition under the inflammation model, placed in 5% CO 2 , Cultivate in a 37°C incubator for 24 hours. Aspirate the cell supernatant for the determination of the expression level of IL-6 secreted by the cells;
[0176] Step b, measuring the concentration of the inflammatory factor IL-6 in the cells: measuring the content of IL-6 in the cell supernatant by PCR method, a...
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